poster abstract numerical index 58 pages

Poster Abstract Numerical Index

(LBR next to the Abstract # indicates "Late-Breaking Research")

Abstract # Presenter Abstract Title Abstract Category & Presentation Day

001 Eric Kmiec, PhD

Targeted Nucleotide Exchange in the CAG Repeat Region of the Human HD Gene

Human Experimental Therapeutics/Monday, August 18

002Tatiana Zaldivar Vaillant, MD Huntington Disease in Cuba Other/Sunday, August 17

003 Laure Jamot, PhDA Strategy for Drug Discovery in Huntington's Disease

Animal Model Experimental Therapeutics/Monday, August 18

004Ferdinando Squitieri, MD, PhD

Homozygosity For CAG Mutation in Huntington Disease is Associated With a More Severe Clinical Course Pathogenetic Mechanisms/Sunday, August 17

005 Reinier Timman, MA

Adverse Effects of Predictive Testing for Huntington Disease Underestimated: Long Term Effects 7-10 Years After the Test Psychosocial Issues/Sunday, August 17

006 Karen Forrest, MLitt

Informing Relatives About Genetic Risk: A Qualitative Study of Patients At Risk For Huntington's Disease or Hereditary Breast/Ovarian Cancer Psychosocial Issues/Sunday, August 17

007Nellie Georgiou-Karistianis, PhD

The Effect of Huntington's Disease on the Ability to Respond to Conflicting Spatial Stimuli Other/Sunday, August 17

008Monique Losekoot, PhD

European Molecular Genetics Quality Network: External Quality Assessment for the Molecular Diagnosis of Huntington Disease in Europe (1997-2002) Genetics/Sunday, August 17

009 Anne Norremolle, PhD

Monozygotic Twins Displaying Mosaicism of an Expanded and an Intermediate Allele of the CAG Repeat in the Huntington Disease Gene Genetics/Sunday, August 17

010Oliver Quarrell, MD, FRCP

UK Study of Reduced Penetrance Alleles in Huntington's Disease Genetic Counseling/Sunday, August 17

011 Wang Jin, PhD

Small Compounds That Inhibit Mutant Huntingtin Aggregation: Potential Therapeutics of HD Patients Other/Sunday, August 17

012Fiona Richards, Master of Social Work

Reproductive Decision Making Before and After Predictive Testing for Huntington Disease: An Australian Perspective Genetic Counseling/Sunday, August 17

013 Marieta Anca, MD

An Abnormal Sleep Pattern and Excessive Movements Can Characterize Patients With Huntington's Disease Other/Sunday, August 17

014 Karen Keast, BSc

Innovative Approaches to Improving the Acceptance of Texture Psychosocial Issues/Sunday, August 17

015Silvia Gines-Padros, PhD Biochemistry

Specific Progressive cAMP Reduction Implicates Energy Deficits in Presymptomatic HD Knock-in Mice Cell Dysfunction/Sunday, August 17

016 Raphael Bonelli, MD

Subcortical Mild Cognitive Impairment: A Novel Concept of Mental Dysfunction Clinical Characteristics/Monday, August 18

017 Craig Bailey, PhD

Genetic Disruption of Tissue Transglutaminase Delays Disease Progression in a Mouse Model of Huntington's Disease Pathogenetic Mechanisms/Sunday, August 17

018 Hans Claus, MSC

Nature and Development of HD in a Nursing Home Population: The Further Development of the BOSH Rating Scale Clinical Characteristics/Monday, August 18


Predictive Testing in Huntington Disease Families With Patients Homozygous for CAG Mutation Genetic Counseling/Sunday, August 17


Study of Juvenile Huntington Disease Patients of Italian Origin Genetics/Sunday, August 17

021Roslyn Tassicker, Master of Social Work

Non-Consensual Predictive Testing: Testing at 25% risk-Some Legal and Counseling Issues Encountered in Australia Genetic Counseling/Sunday, August 17

022Maurice Curtis, BhSc, MSc

Increased Cell Proliferation and Neurogenesis in Huntington's Disease


023Paola Zinzi, psychology

Effects of Rehabilitation on Motor Performance, Mood State and Everyday Life of HD Patients Psychosocial Issues/Sunday, August 17

024Jeroen van Vugt, MD, PhD

Objective Assessment of Akinesia and Bradykinesia in Huntington's Disease Clinical Characteristics/Monday, August 18

025 Lilias Barron, BSc

Comparisons of CAG Repeat Distributions for Familial and Sporadic HD Cases & Intermediate Allele Frequencies For Different Populations Support a Stepwise Model for CAG Expansion Other/Sunday, August 17

026 David Blum, PhD

A Dual Role of Adenosine A2A Receptors in 3-Nitropropionic Acid-Induced Striatal Lesions: Implications for the Neuroprotective Potential of A2A Antagonists

Animal Model Experimental Therapeutics/Monday. August 18

027 Ronald Risley, MD

Multidisciplinary Care in the Information Age a System for Coordination of Care Amongst Geographically Dispersed Providers Other/Sunday, August 17

028 Vicki Wheelock, MD

Somatoform Disorders as the Initial Presentation of Huntington Disease Clinical Characteristics/Monday, August 18

029 Alis Hughes, Ms

The Huntingtin-Interacting Protein Endophilin A3 Forms Filamentous Structures Which Associate With Microtubules But Not With Actin Filaments Other/Sunday, August 17

030Christopher Kipps, MBBS

Progression of Structural Neuropathology in Huntington's Disease Using Voxel and Tensor Based Morphometry Neuroimaging/Monday, August 18

031 Neil Mahant, MBBS

Does the Clinical Spectrum of Huntington's Disease Include Tremor Clinical Characteristics/Monday, August 18

032Krisna Vaddadi, FRC Psych

Treatment of Huntington's Disease Using Essential Fatty Acids (Long Term Follow Up Case Presentation)


033 Din-E Shan, MD, PhD

Stereotypic Leg Movements in a Family Member of Huntington's Disease Clinical Characteristics/Monday, August 18

034Marie-Noelle Witjes-Ane, MSc

Psychomotor Slowing and Memory Disturbances Over a Three-Year Period in Presymptomatic' carriers for Huntington's Disease Clinical Characteristics/Monday, August 18

035 Julie Snowden, PhD

Assessment of Change in Cognitive Function in Huntington's Disease Clinical Characteristics/Monday, August 18

036 Nazli Basak, PhDMolecular Analysis of Turkish Huntington's Disease Patients Genetics/Sunday, August 17

037 Nagehan Ersoy, MSc

Interaction Between Huntingtin and the Nuclear Receptor Corepressor

Cellular-Based and Genetic Animal Models/Monday, August 18

038 Bjarke Naver, MSc

PEI Mediated In Vivo Gene Transfer in Transgenic Huntington's Disease Mice. Evaluation of Transfection Method, Effect and Biomarkers


039Lis Hasholt, Dr. Med.Sci.

Abnormal Morphology of HD Fibroblasts Grown in Enriched Culture Medium Cell Dysfunction/Sunday, August 17

040 Deborah Russel, BA

High Throughput Screen To Identify Compounds That Downregulate Intracellular Mutant Huntingtin Expression Other/Sunday, August 17

041Yvette Grimbergen, MD

Does the Cause of Functional Disability Change With Disease Progression in Patients with Huntington's Disease? Clinical Characteristics/Monday, August 18

042 Erica Johnson, BS

Identification in Cell Culture of Small, Recruitment Positive Polyglutamine Aggregates: Aggregation Foci


043 Hoon Ryu, PhD

Histone Deacetylase Inhibitors Prevent Oxidative Neuronal Death Independent of Expanded Polyglutamine Repeats via a Sp1-Dependent Pathway


044

Alison Lashwood, MSc. Genetic Counseling

Preimplantation Diagnosis for Huntington Disease-The London Experience Genetic Counseling/Sunday, August 17

045Sheila Simpson, MBChB, BSc, MD

PEG Feeding In End Stage Huntington's Disease: An Examination of the Clinical and Ethical Issues, Including the Views of the Doctor, Patient and the Patient's Family Other/Sunday, August 17

046 Anita Bruce, BSc

The Impact of Presymptomatic Predictive Testing for Huntington's Disease on Partners and Marital Relationships: Secrets, Lies and Truths Genetic Counseling/Sunday, August 17

047Teresa Tempkin, RNC, MSN, ANP

Improving Psychiatric Outcomes in an Outpatient HD Population Psychosocial Issues/Sunday, August 17

048David Craufurd, FRCPsych

Changes in Sexual Behaviour Associated with Huntington's Disease Clinical Characteristics/Monday, August 18

049David Craufurd, FRC Psych

Measurement of Irritability in Huntington's Disease


050

Elizabeth Howard, BSc, MB ChB, MRCGP

Walk and Drink Time; Objective Measures of Motor Progression in Huntington's Disease Clinical Characteristics/Monday, August 18

051 Heike Goehler

Analysis of Huntingtin Protein-Protein Interactions Using the Yeast Two-Hybrid System Pathogenetic Mechanisms/Sunday, August 17

052Collene Lawhorn, BS, MS Ed

Motor Deficits and Striosome Volume in the YAC72 Mouse Model of Huntington's Disease Other/Sunday, August 17

053 Gary Dunbar, PhD

Use of Adult Bone Marrow Stem Cell Transplants to Counteract Cognitive Deficits in a Rat Model of Huntington's Disease


054 Hui Zhu, PhD

Toxicity of a Carboxyl-Terminal Huntingtin Fragment in Drosophila


055Jennifer Thompson, B.Sc

Social Cognition in Huntington's Disease and Frontotemporal Dementia Clinical Characteristics/Monday, August 18

056 Heloisa Ruocco, PhD

Quantification of Caudate and Putamen Atrophy by Volumetric MRI in Patients with Huntington Disease. Association Between Length of the CAG Repeat and Age at Onset Neuroimaging/Monday, August 18

057Wen-Qi Zeng, MD, PhD

Sequence and Haplotype Analysis of GRIK2 (GluR6) in Huntington Disease: Assessment of a Modifier Gene Genetics/Sunday, August 17

058 Birgit Zucker, MD

A Quantitative Gene Expression Analysis Of Laser-Dissected Striatal Neurons In The R6/2 Mouse Model Of Huntington's Disease Cell Dysfunction/Sunday, August 17

059Marguerite Wieler, BScPT

Brain Energy Metabolism in Huntington's Disease Neuroimaging/Monday, August 18

060Elisabeth Almqvist, PhD, RN

Personality Traits in Probands Undergoing Predictive Testing for Huntington Disease Genetic Counseling/Sunday, August 17

061 Sarah Augood, PhD

mRNA Profiling of Two Striatal Neuron Populations with Differential Vulnerability in HD: A Study Utilizing LCM, Real-Time PCR, and cDNA Microarrays

Neuronal Vulnerability and Survival/Monday, August 18

062Ellizabeth McCusker, MBBS, FRACP

Parenting Patterns in Patients with Huntington's Disease Psychosocial Issues/Sunday, August 17

063 Blair Leavitt, MD, CM

Ethyl-Eicosapentaenoate (Ethyl-Epa) in Huntington's Disease: A Randomised, Placebo-Controlled Trial


064Parsa Kazemi-Esfarjani, B.Sc., PhD

Suppression of a Polyglutamine Toxicity by a Drosophila Ortholog of MRJ



MRI and PET Assessment of Brain Involvement in Subjects With Presymptomatic, Initial and Advanced Huntington Disease Neuroimaging/Monday, August 18

066 Kaori Muto, PhDWho Killed That Lady? A HD Murder Case in Japan Psychosocial Issues/Sunday, August 17

067 Raphael Bonelli, MD

Management of Huntington's Disease: An Evidence-Based Review


069 Donald Higgins, MDDefining Progression in Huntington Disease Clinical Characteristics/Monday, August 18

070Elise Kayson, MS, RNC

Safety and Feasibility of the Prospective Huntington At Risk Observational Study (PHAROS): A Progress Report Clinical Characteristics/Monday, August 18

071Fabiola Hormozian, Ms.c in Genetics

Molecular Study of Huntington Disease in Iranian Families Genetics/Sunday, August 17

072 Adam Rosenblatt, MD

Clinical predictors of neuropathological severity: a prospective autopsy study of 100 individuals with Huntington's disease Clinical Characteristics/Monday, August 18

073 Ayana Duckett, BA

Risk factors for Osteoporosis in Women with Huntington's Disease in a Long-term Care Setting Clinical Characteristics/Monday, August 18

074Wance J.J. Firdaus, DEA, MSC

Heat Shock Protein 27 Protects Monkey Cos Cells Against Oxidative Stress Generated by the Expression of Mutant Huntington Exon 1 Cell Dysfunction/Sunday, August 17

075 George Lawless

Huntington Expression Results in a Decrease of Proteasome Activity and Triggers Apoptosis in PC12 Cells Cell Dysfunction/Sunday, August 17

076 Elena Moro, MD, PhDBilateral GP Stimulation for Huntington's Disease Other/Sunday, August 17

077 Lisa Ellerby, PhDThe Role of Calpain in Huntington's Disease


078-LBR Michel Cyr, PhD

Behavioral Manifestations of High Extracellular Dopamine in HdhQ92 Knock-in Mice


079 Carsten Saft, MDPeg Insertion Reflects Dysfunction in Huntington's Disease Clinical Characteristics/Monday, August 18

080-LBR Carsten Saft, MD

Is 41 Better Than 45? Influence of Low CAG Ranges on Huntington's Disease Clinical Characteristics/Monday, August 18

081-LBR Kevin Biglan, MD

Emerging Cohort Characteristics in the Prospective Huntington At Risk Observational Study (PHAROS) Clinical Characteristics/Monday, August 18

082 Kristy Bolter, PhDImplicit Processes in People with Huntington's Disease Clinical Characteristics/Monday, August 18

083-LBR Charity Aiken

Cell Culture Screen Identifies Caspase Inhibitors as Potential Drugs for the Treatment of Huntington's Disease


084-LBRElizabeth Aylward, PhD

Onset and Rate of Change in Basal Ganglia Volume in Presymptomatic Subjects Neuroimaging/Monday, August 18

085 Alonso Montoya, MD

Episodic Memory Impairment in Huntington's Disease: a Meta-Analysis Clinical Characteristics/Monday, August 18

086-LBR Yona Gefen, PhD

Cop-1 Vaccination Reduces Motor Function Deficits and Increases Life Expectancy in a Transgenic Mouse Model of Huntington's Disease


087-LBR Irina KovtunMsh2 Protein in Base-excision Mediated CAD Expansion Pathogenetic Mechanisms/Sunday, August 17

088Kathleen Clarence-Smith, MD, PhD

Mechanism of Action of the Anti-Chorea Drug tetrabenazine: A Review


089-LBR Jane Paulsen, PhD

Initial Baseline Characteristics in the PREDICT-HD (Neurobiological Predictors of HD) Study Clinical Characteristics/Monday, August 18

090 Francois Richer, Phd

Frontal Hypoactivation and Reduced Voluntary Control in Early Huntington's Disease Neuroimaging/Monday, August 18

091 Francois Richer, Phd

Pupilary Measure of Brainstem Activation is Reduced in Huntington's Disease Clinical Characteristics/Monday, August 18

092 Jessica Easton, M.A.

Self-Understanding and Identity: The Experience of Adolescents At Risk for Huntington's Disease Psychosocial Issues/Sunday, August 17

POSTER ABSTRACT SESSION #1SUNDAY, AUGUST 17, 2003

7:45 a.m. – 8:45 a.m.

All Congress posters will be on display Sunday and Monday.

We request poster presenters to stand by their poster during poster abstract session #1 on

Sunday, August 17 7:45 a.m. to 8:45 a.m.

The following poster topics have been assigned to poster session #1:

Genetics, Genetic Counseling, Psychosocial Issues,

Pathogenetic Mechanisms, Cell Dysfunction,

Other

(Abstracts for the above topics to follow…)

TOPIC: GENETICS

ABSTRACT 008: European Molecular Genetics Quality Network: External Quality Assessment for the molecular diagnosis of Huntington Disease in Europe (1997-2002).

Monique Losekoot, Bert Bakker (DNA-diagnostic Laboratory, Centre for Human and Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands), Su Stenhouse (Northern Genetics Knowledge Park, Bioscience Centre, International Centre for Life, Newcastle, UK), Valerie Biancalana (CNRS INSERM, University Louis Pasteur, Institute of Biological Chemistry, Strasbourg, France), Franco Laccone (Institute of Human Genetics, Universitaet Goettingen, Goettingen, Germany), Simon Patton, Rob Elles (European Molecular Genetics Quality Network, National Genetics Reference Laboratory, Manchester, UK)

The European Molecular Genetics Quality Network (EMQN) was established with the aim of raising and maintaining standards of Clinical Molecular Genetic Testing in the EU through the provision of standard External Quality Assessment (EQA) schemes and agreed Best Practice protocols. Huntington Disease (HD) is one of the diseases for which EQA schemes are offered by the EMQN. An EQA scheme on HD has been run from 1997-now on a yearly basis. Three DNA samples, matched to mock clinical referrals, were sent out and analysed by the participating laboratories. Genotyping results as well as reports in normal laboratory format (in English) were assessed on an anonymous basis on previously agreed marking criteria by a panel of 3 assessors from different countries. Individual comments were returned to the participants together with the scores on genotyping and interpretation of the results in the light of the clinical data supplied. A report containing a general overview of the scheme was provided. Between 14 and 50 laboratories from more than 22 different countries participated over the years. The EQA schemes demonstrate a level of misdiagnosis: with the exception of 2002, 1 to 3 diagnostic errors (1-3% of cases) were made each year. In contrast, an increasing number (55% in 1998 to 80% in 2002) of the participants typed all alleles within the error limits. The Interpretation (laboratory reports) was more difficult to assess, but there is an overall trend towards a more uniform way of reporting. The results of the schemes will be presented and discussed. The errors identified indicate a clear need for EQA to measure current standards of proficiency and encourage laboratories to raise their technical performance.

ABSTRACT 009: Monozygotic twins displaying mosaicism of an expanded and an intermediate allele of the CAG repeat in the Huntington Disease gene.

Anne Nørremølle, Lis Hasholt, Cathrine B. Petersen, Hans Eiberg and Sven Asger Sørensen (Dept. of Medical Genetics, University of Copenhagen, Denmark)

We here present the first case of a monozygotic twin pair with mosaicism for different alleles of the CAG repeat in the Huntington Disease gene. The twins are 34 years of age; the firstborn twin is clinically normal whereas the second twin shows clear signs of HD with onset around 30 years of age. PCR amplification of the CAG repeat sequence in DNA from leucocytes and buccal epithelial cells revealed three products of different size in each of the twins, corresponding to a 20 CAG normal allele, a 37 CAG intermediate, and a 47 CAG HD allele. The intermediate allele had arisen through mitotic contraction of the expanded allele inherited from the affected mother. Analysis of DNA from single hair roots showed only two alleles: the normal allele and one of either the intermediate or the expanded allele, confirming mosaicism. All three alleles are transcribed. The proportion of the two cell populations differs between the twins in both skin fibroblast cultures, hair roots and sperm samples. Of special interest are the analyses of hair roots, which like CNS are of ectodermal origin; here the affected twin revealed only the normal and the expanded allele. Sperm cells from the non-affected twin carried the normal or a 37 CAG allele, whereas sperm cells from the affected twin carried the normal allele or an expanded allele with 47 to >70 CAG’s. Difference in age at onset among monozygotic HD twins has been reported (Georgiou et al. Mov Disord. 14:320-5, 1999); it is still too early to make a statement on the clinical differences in the present twins. Cultured cells from the twins constitute a remarkable cell model for studies of the cellular pathogenesis due to expression of an intermediate and an expanded HD allele with identical genetic background.

ABSTRACT 020: Study of juvenile Huntington disease patients of Italian origin

Milena Cannella, Vittorio Maglione, Patrizia Giallonardo, Ferdinando Squitieri (Neurogenetics Unit, IRCCS Neuromed, Localita’ Camerelle, 86077, Pozzilli, IS, Italy).

We analyzed a population of juvenile Huntington disease (HD) subjects of Italian origin (n = 57). The main aim of this study was to analyze the gender effect of the affected parent on age at onset and clinical presentation of offspring with juvenile HD. We also analyzed molecular features of the disease, including CAG mutation length and GluR6 gene polymorphism, according to the affected parent’s gender. The mutation length was longer in paternally than in maternally transmitted HD juvenile patients (p = 0.025), nevertheless a similar mean early onset in the two groups (p > 0.05). This data was even enforced by that obtained from the whole cohort of patients included in the databank (n = 600) where, in the presence of increased mean parent-child CAG repeat change in paternal vs maternal meiotic transmissions (+ 7.3 vs + 0.7 CAG, p = 0.0002), the mean parent-child year-of-onset change was similar in the two groups (-10.4 and –7.0 years, p > 0.05). A lower TAA-triplet in GluR6 was associated with an earlier age at onset in juvenile patients (p = 0.031, R2 = 0.10). When we added the GluR6 effect on age at onset to the CAG expanded number effect (p = 0.0001, R2 = 0.68) by multiple regression approach, the coefficient of determination R2 increased to 0.81. This effect in addition to the expanded CAG repeat number, found in juvenile and not in adult patients, was slightly enforced by paternal compared to maternal transmissions (R2 0.82). Our findings suggest the occurrence of a weaker effect of the paternal mutation on juvenile age at onset in our population, possibly amplified by other genetic factors, such as the TAA-triplet length in the GluR6 gene.

ABSTRACT 036: MOLECULAR ANALYSIS OF TURKISH HUNTINGTON’S DISEASE PATIENTS

Nagehan Ersoy and A. Nazli Basak Bogazici University, Department of Molecular Biology and Genetics, Istanbul, Turkey

Routine genetic testing of Huntington’s Disease (HD) was initiated at our center five years ago by direct mutation analysis. During this period, a total of 65 individuals presented in our department requiring molecular analysis. Forty four of these individuals were clinically diagnosed with HD, eleven were presymptomatic, and the remaining ten required differential diagnosis. Genetic counselling and informed consent was obtained in all cases. The method used for genetic testing involved extraction of DNA from leukocytes, PCR-amplification of the CAG-repeat region, electrophoresis of PCR products on a denaturing polyacrylamide gel and visualization by silver staining. Allele sizing was done on a calibration curve. Using this approach, clinical diagnosis was confirmed in 40 of the 44 clinically diagnosed individuals, the remaining four did not carry a CAG expansion in their HD genes. Eight out of eleven presymptomatic individuals had a favorable result, whereas three of them carried an HD allele. Out of ten patients requesting differential diagnosis, one proved to have an expanded allele. All patients exhibited characteristic clinical symptoms; e.g. behavioral changes, choreic movements and cognitive impairment. Repeat numbers on disease alleles varied between 40 and 58, the most frequent allele being 44. The age of onset of the patients was found to be inversely proportional to the repeat length. Follow-up of the patients and presymptomatic individuals is expected to reveal progression and onset of disease manifestations.No personal financial relationship

ABSTRACT 057: Sequence and haplotype analysis of GRIK2 (GluR6) in Huntington Disease: assessment of a modifier gene

Wen-Qi Zeng, Richard Myers, Marcy MacDonald, James F. Gusella (Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129 and Department of Genetics, Harvard Medical School)

In Huntington disease, only 50%-60% of the variation in onset age can be explained by the size of the CAG repeat expansion. Previous results have showed a significant association of a particular allele of GRIK2 (GluR6) - UTR TAA repeat polymorphism with age of onset, suggesting it might be in linkage disequilibrium with a functional variant in the GRIK2 gene, which encodes a subunit of the glutamate receptor. Four pairs of patients with similar CAG repeats but quite different ages at onset were selected for sequencing of the GRIK2 coding regions and subsequently for haplotype analysis. 17 exons and their splice junction regions were sequenced and compared. There was no coding sequence variant in GRIK2 that explained the effect on age at onset in these cases, suggesting a regulatory difference or an effect due to an adjacent gene. 10 microsatellite markers were chosen in the GRIK2 region and genotyped in the paired samples for haplotype analysis. There was no evidence for a single ancestral haplotype for the GRIK2 alleles associated with earlier than expected HD onset. Sequence analysis of the 17;UTR regions suggested that the TAA alleles did not share a common origin and therefore might themselves be responsible for the observed effect, presumably through a regulatory effect at the RNA level. This possibility is now being explored.

ABSTRACT 071: Molecular study of Huntington disease in Iranian families

Fabiola Hormozian,Mohammad M Banoei,Massoud Hushmand,Mohammad H sanati (NRCEGB, POBox:14155-6343 Tehran/Iran)

BACKGROUND Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder associated with expansions of an unstable CAG trinucleotide repeats in exon 1 of the IT15 gene. Located on chromosome 4p16.3. Choreiform movements and cognitive impairment characterize it. Onset of symptoms is around 40 years of age and progression to death occurs in approximately 10 to 15 years from the time of disease onset. In this study we present the first molecular data on the basis and the origin of Huntington disease in Iran.

PATIENTS AND METHOD: To obtain molecular data, genomic DNA from 51 patients and asymptomatic at risk members belongs to 20 affected families and 28 normal control were amplified in the involved region by polymerase chain reaction. The CAG repeat numbers varied from 40 to 74 (median: 52.6) in HD patients and asymptomatic carriers, while individuals of the normal control group had 7-34 CAG repeat numbers (median: 17.9). In our study age of onset varied from 17 to 56. We observed a significant increase of repeat size for paternal transmission of the disease and greater instability for paternally transmitted CAG repeats in the HD size range There were large CAG repeats (74 copies) in paternally transmitted HD case with early onset (age 17).

CONCLUSION: Data generated from this study may have significant implications for the etiology, knowledge of the incidence, diagnosis, prognosis, genetic counseling and treatment of HD Iranian patients. Therefore, molecular confirmation of the clinical diagnosis in HD should be sought in all suspected patients, making it possible for adequate genetic counseling.

TOPIC: GENETIC COUNSELING

ABSTRACT 010: UK Study of Reduced Penetrance Alleles in Huntington’s Disease

Oliver W Quarrell (Dept. Clinical Genetics, Sheffield Childrens’ Hospital, Sheffield UK)Alan S Rigby (Unit of Medical Statistics, University of Sheffield, Sheffield UK) Members UK HD Consortium

Four types of predictive test results may be recognised: <26 repeats is unequivocally normal; 27-35 repeats is normal but in a range which may give rise to abnormal expansions in future generations; 36-39 repeats is abnormal but the individual may or may not develop the condition; 40 or more repeats is unequivocally abnormal(1). There are no empirical risk figures for individuals with a result in the 36-39 range(1). A survey of UK laboratories offering mutation analysis for Huntington’s disease indicated that there had been at least 170 results in this range from both pre-symptomatic and diagnostic tests.

Ethical committee approval has been obtained to allow anonymous collection of data on age of onset or age last known to be asymptomatic for each case with a result in the reduced penetrance range. As part of the study, DNA samples will be re-analysed. All 21 UK centres have agreed to participate in the study.

Information collection started in Feb 03; so far, data is available on 41 cases: 26 are affected with ages of onset ranging between 38-80 years; 15 cases are asymptomatic with ages ranging between 35-71 years; 16 of the 41 cases are or would have been asymptomatic at the age of 60 years. Data collection will continue for the rest of the year.

The latest data will be presented at the meeting together with a preliminary Kaplan-Meier curve.1. ACMG/ASHG Statement Laboratory guidelines for Huntington disease genetic testing Am J Hum Genet 1998; 62: 1243-1247.

ABSTRACT 012: Reproductive Decision Making Before and After Predictive Testing for Huntington Disease: An Australian Perspective

Fiona H Richards (Department of Clinical Genetics, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, 2145, Australia) Gillian Rea (Faculty of Medicine, University of Aberdeen, Polwarth Building, Foresterhill, Aberdeen AB25 2ZD, UK)

This study, an update of an unpublished 1997 study, retrospectively examines the reproductive decisions of 373 adults who underwent predictive testing for Huntington disease (HD) in Sydney, Australia, between 1990 and 2002. Data were collected from genetics records and follow up contact.The total group consisted of 175 males and 198 females (mean age = 38 years, range 18 - 82 years). Of these, 222 (59.5%) had one or more pregnancies prior to predictive testing. In three pregnancies prenatal (linkage) tests were performed. Two were low risk, and one high (50%) risk pregnancy was terminated. Four couples utilised alternative reproductive options prior to predictive testing; one each of (overseas) adoption, fostering, IVF/donor egg, and donor sperm. Post-result analysis was performed for a subgroup of 231 persons in the reproductive age group of 18 to 45 years, for whom follow up data were available (mean age = 33.2, sd = 6.8). 109 (47%) were found to be carriers of the HD mutation* and 122 (53%) were non-carriers. 30 carriers (28%) and 39 non-carriers (32%) had one or more post-result pregnancies. This difference was not significant ( = .543, p = .461).

Five of the carriers (17%) who had a post-result pregnancy had a total of six prenatal tests - two results were non-carrier, and 4/4 carrier result pregnancies were terminated. 24 carriers (22%) were symptomatic at the time of testing - three (12.5%) of these had a post-result pregnancy. One pregnancy was terminated and the other two continued, all without prenatal testing. Two carriers utilised alternative reproductive options; one each of donor sperm and preimplantation genetic diagnosis. One non-carrier adopted two children and another non-carrier fostered two children.

These results support previous research that reports a low uptake of prenatal testing and other reproductive options.Included 15 whose result was in the intermediate range with risk to offspring. The authors have no personal financial relationship with any products or sponsors associated with the World Congress on Huntington’s Disease.

ABSTRACT 019: Predictive testing in Huntington’s disease families with patients homozygous for CAG mutation

Ferdinando Squitieri, Milena Cannella, Maria Simonelli (Neurogenetics Unit, IRCCS Neuromed, Localita’ Camerelle, 86077, Pozzilli, IS, Italy) Elisabeth W. Almqvist (NeuroHealth Consulting Sweden HB Hemslöjdsvägen 8, 3 tr S-167 31 Bromma, Sweden) Giuliana Cislaghi (Fornaroli Hospital, 20013, Magenta, MI, Italy). Cinzia Gellera, Stefano Di Donato (Istituto Neurologico Besta, via Celoria 11, 20133, Milan, Italy). David C. Ribinsztein (Cambridge Institute for Medical Research, Welcome Trust/M.R.C. BLD6 Addenbrooke’s Hospital, Cambridge, England CB2). David Turner (Department of Haematology and Genetic Pathology School of Medicine Flinders University, Adelaide, Australia 5042).Anne-Catherine Bachoud-Lévi (Equipe Avenir, INSERM U421, Faculté de Medicine, Paris XII and Service de Neurologie, Hopital Henri Mondor, Creteil, France). Sheila A. Simpson (Department of Medical Genetics, Grampian University Hospitals Trust, Aberdeen, Scotland). Martin Delatycki (Victorian Clinical Genetics Services, The Murdoch Institute, Royal Children’s Hospital, Melbourne, Australia). Yuval O. Herishanu (Department of Neurology, Soroka Medical Center, Ben-Gurion University of the Negev, P.O.Box 151, Beer-Sheva 84101, Israel). Marie-Francoise Chesselet (Department of Neurology, The Geffen School of Medicine at UCLA Reed Neurological Research Center, B114 - 710 Westwood Plaza Los Angeles, CA 90095, USA). Michael R. Hayden (Centre of Molecular Medicine and Therapeutics, Department of Medical Genetics, University of British Columbia, 980 West 28 avenue, room 302y Vancouver, BC, V5Z4H4 Canada).

Huntington disease (HD) is caused by a dominantly transmitted CAG repeat expansion mutation that is believed to confer a toxic gain-of-function on the mutant protein. HD patients with two mutant alleles are very rare. In other poly(CAG) diseases such as the dominant ataxias, inheritance of two mutant alleles causes a phenotype more severe than in heterozygotes. As this genetic condition is rare and its implication with the severity of HD phenotype has never been pointed out so far, it has never been contemplated by the predictive testing (PT) programs. It would therefore be of relevance for international guidelines and Predictive Testing programs to document the existence of unusual genetic conditions (i.e. subjects with HD homozygosity, 25% risk or premutation carriers), which may require different strategies in the genetic counselling approach according to the possible diverse scenarios occurring within the families, mainly where the parents’ consanguineity may generate HD homozygosity: 1. Increased risk of homozygotes’ offspring to inherit the HD mutation (100% instead of the usual 50%); 2. Increased risk of offspring with both parents heterozygous for HD mutation to inherit the HD mutation in either a homozygous (25%) or heterozygous (50%) condition (total risk of inheritance of 75%). We documented the existence of 14 subjects homozygous for CAG mutation. Two of them were unaffected and, therefore, in a presymptomatic stage of life while some of their relatives had 100% risk to be mutation carriers (homozygotes’ offspring) or a potential risk to be mutation carriers of 75% (homozygotes’ siblings), higher than the usual 50% risk to inherit the mutation. Two patients came from families from small areas/communities of Norther Italy and Israel, where marriages occurred among relatives potentially contributing to HD clusters. We are studying DNA polymorphisms in markers close to the 5’ CAG mutation (CCG, Del 2642, CA repeat, D4S227) in five homozygotes to analyse the potential within-family founder effect of the mutations in homozygotes’ families. Preliminary results indicate that at least the intragenic markers are themselves homozygous in these subjects suggesting an identical founder effect of both mutations within each family. We set up a patients’ tissue bank and are currently working on peripheral cells obtained from subjects homozygous and heterozygous for HD mutation to look at potential biological differences in these two groups of patients.

ABSTRACT 021: Non-consenusal predictive testing: testing at 25% risk - some legal and counselling issues encountered in Australia

Roslyn J Tassicker, Genetic Health Services Victoria, Royal Children’s Hospital, Flemington Road, Parkville 3052 Vic. Australia

Does the clinician have a legal or clinical responsibility to a person who has not contacted the Genetics Service? Two requests for prenatal testing to this Service were made by pregnant women whose male partners were at 50% risk. The male partners were not aware prenatal testing was being sought. Legal opinion was obtained regarding testing *. This poster summarises further clinical issues.International Guidelines prioritise the right of an individual at 25% risk of having the HD mutation to access testing over the right of their parent to not know. Both parties are encouraged to discuss testing. There is implied responsibility of the clinical team to the person at 50% risk whose genetic status may be indirectly revealed.

While some European experiences of testing individuals at 25% risk have been documented, numerous contentious counselling and legal matters remain unclear. A survey across 5 Australian states on testing those at 25% risk reveals Counsellors consider the following:

o is contact between parent and child ongoing?, is the parent aware testing is being sought?, the nature of the relationship., is the parent symptomatic but undiagnosed?

o time frames and contingent decision making.

Two (assumed monozygotic) twin cases have arisen in which one twin wanted testing and the other did not. In both cases, legal advice was that testing should proceed for the requesting twins, and the non-requesting twins should be warned of possible harm and encouraged to seek support of Genetic Services.Conclusion: Clinicians legal responsibility to an at-risk individual who has not contacted a Service is unclear. Counsellors may still reflect consideration for this individual in their practice. *Tassicker, R. et al, BMJ, 326: 331-333, 2003

ABSTRACT 044: Preimplantation Diagnosis for Huntington Disease- The London Experience

Lashwood Alison, Black Cheryl, Abbs Stephen, Renwick Pam, Flinter Frances, Braude Peter (Centre for Preimplanatation Diagnosis, Guy's & St. Thomas' Hospital, London).

Preimplantation genetic diagnosis (PGD) combines in vitro fertilisation with molecular analysis of a single blastomere biopsy. PGD for Huntington disease (HD) is available in our centre for those couples where one partner carries the HD expansion mutation. In accordance with the UK Human Fertilisation & Embryology Act (1990), a licence to practise PGD is required. There are several issues relating to PGD for HD that require special attention; we have addressed these by developing a detailed working protocol and we report our experience with this to date.

Prospective patients undergo extensive counselling to discuss the process of PGD and, for HD, we consider the welfare of any children born into a family where one parent will become symptomatic. Following counselling, eighteen couples requested preparatory molecular work up for HD PGD. To be informative for PGD all 4 parental allele sizes must be known, which requires the unaffected partner to undergo a modified presymptomatic test. The accuracy and likely success with PGD is greater if a couple is fully informative, i.e. both allele sizes of the unaffected partner are different from the normal allele size in the affected partner. Thirteen of 18 couples were fully informative. Seven cycles of PGD have been completed in 4 fully informative couples. Two cycles failed to reach embryo transfer. Up to 2 embryos were transferred in 5 cycles resulting in 3 pregnancies. One triplet pregnancy miscarried, one twin pregnancy resulted in live born twins and one singleton pregnancy is ongoing. Despite a misdiagnosis risk of 2-5% per embryo, all couples decided against confirmatory prenatal diagnosis (PND), and under our protocol no confirmatory testing is possible at birth. We believe that, despite its complexity, PGD HD offers an acceptable alternative to routine PND and our protocol sets a standard for PGD for other late onset disorders.

ABSTRACT 046: The impact of presymptomatic predictive testing for Huntington’s disease on partners and marital relationships: secrets, lies and truths.

Anita M Bruce, June Semper, Sheila A Simpson (Grampian University Hospitals Trust, Aberdeen, Scotland, UK)

Three classic clinical features are associated with Huntington’s Disease (HD): movement disorder, personality changes and cognitive impairment. This autosomal dominant neurodegenerative disorder usually develops between the ages of 30 and 50. For most people this is considered as the prime of life, when social responsibilities and personal and financial possibilities are greatest. Individuals have often married and had children before they had any knowledge of the family history of HD or their own risk, and this is generally considered to be one of the most distressing issues of this disease.

Individuals with HD exhibit a high degree of personality and behavioural changes. The spouse has to cope with their deteriorating partner’s physical needs and disabilities and also with slowly losing a companion and life long partner. Intimacy, friendship and decision making deteriorate to the point when many spouses feel that the individual they are caring for is no longer the person they married. Accurate presymptomatic testing has been available since 1993, but studies tend to focus on the person at risk, rather than their partner. The partner is sharing the life of the person at risk, and must be empowered during the decision making process and given care and support after the result is given. We have investigated the effects of presymptomatic testing for HD on the marriages and intimate relationships of 39 couples, and have explored the additional effect of having kept secret a family history of HD prior to marriage. The partners require the opportunity to have good communication with the person at risk and the clinicians dealing with them, regardless of whether favourable or unfavourable results are produced.

ABSTRACT 060: Personality traits in probands undergoing predictive testing for Huntington Disease

Elisabeth W. Almqvist, Petter Gustavsson, (Dept. of Nursing, Karolinska Institute) Ulla Platten, Aina Haegermark, Urban Ösby, Maria Anvret, (Dept. of Clinical Genetics, Karolinska Hospital) Tarja-Brita Robins Wahlin, (Neurotec, Karolinska Insititute), Anders Lundin, (Dept. of Neurology, Karolinska Hospital)

Personality changes are prominent features of Huntington Disease (HD). Shiwach (1994) recorded personality changes in 72% of a sample of 110 HD patients. However, little is known about prodromal signs of personality changes in asymptomatic individuals carrying the HD mutation.

A total of 91 probands undergoing predictive testing for HD at the Karolinska Hospital, Stockholm participated in the study. The questionnaire Karolinska Scales of Personality (KSP) (Schalling et al., 1987) was administered at baseline (pre-test). The KSP is a self-report inventory measuring dimensions of personality, preferentially considered to have a biological basis. The Beck Depression Inventory (BDI) (Beck et al., 1961) was administered at baseline and after test results at 10 days, 2, 6, 12, and 24 months. The aim of the study was to answer the following questions: 1) Do asymptomatic mutation carriers show any prodromal signs of personality change? 2) Are there any personality traits that predispose individuals to apply for the HD predictive testing program? 3) Does personality assessment at baseline reveal vulnerability that predicts adverse events in the follow-up period, as measured by the BDI?

A predictor of adverse events was identified; the Socialization Scale of the KSP (indicating maladaptivity) at baseline was correlated to increased BDI scores during the follow-up period, irrespective test result outcome. However, there was no difference between the mutation carriers and the non-carriers in the personality assessment at baseline. No particular personality traits were seen in the whole study group compared to normal controls. It is of great clinical value to identify predictors of adverse reactions in order to prevent such reactions by extra support during pre- and post-test counselling. References: Beck A et al. (1961) Arch Gen Psychiatr (4): 53-63, Schalling D et al. (1987) Acta Psychiatrica Scandinavica, 76: 172-182, Shiwach R (1994) Acta Psychiatrica Scandinavica 90 (4): 241-6

TOPIC: PSYCHOSOCIAL ISSUES

ABSTRACT 005: Adverse Effects of Predictive Testing for Huntington Disease Underestimated: Long Term Effects 7-10 years after the test

Reinier Timman, Raymund Roos, Aad Tibben (Leiden University MedicalCentre, Department of Neurology, P.O. Box 9600, NL-2300 RC Leiden, The Netherlands), Reinier Timman (Erasmus University Medical Center, Department of Medical Psychology & Psychotherapy, P.O. Box 1738, NL-3000 DR Rotterdam, The Netherlands), Anneke Maat-Kievit (Erasmus University Medical Center, Department of Klinnical Genetics, P.O. Box 1738, NL-3000 DR Rotterdam, The Netherlands), Aad Tibben (Leiden University Medical Centre, Department of Clinical Genetics, P.O> Box 9600, NL-2300 RC Leiden, The Netherlands)

This study assessed the effects of a long-term follow-up of predictive testing for Huntington disease (HD) in The Netherlands. Three questionnaires were administered up to 7 times during a period of 7 to 10 years from 142 individuals at risk for HD who requested a predictive test, accompanied by 104 partners. These questionnaires included future expectancies (Beck Hopelessness Scale), intrusion of adverse ideas and denial-avoidance reactions (Impact of Event Scale) and general well-being (General Health Questionnaire). Carriers and their partners were temporarily more distressed immediately after the test results although their outlooks improved somewhat in the 2-3 year post-test period. However, they became more pessimistic in the long term, when they approached the age of onset. Non-carriers reported strong relief after the test result, but eventually returned towards pre-test levels. Carriers, who did not return questionnaires after they had received their test results, had reported more distress than carriers who both returned questionnaires and came back for additional counseling. This demonstrates that studies reporting few harmful effects among people who had received an unfortunate test result, have underestimated their real impact. Moreover, former studies did not investigate time effects for periods longer than only one to a few years.

ABSTRACT 006: Informing relatives about genetic risk: a qualitative study of patients at risk for Huntington’s disease or hereditary breast/ovarian cancer.

Karen Forrest, Sheila A Simpson, Edwin van Teijlingen, Lorna McKee, Neva Haites, Eric Matthews (University of Aberdeen, Aberdeen, Scotland), Brenda J Wilson (University of Ottawa, Canada).

Background - Recent research has highlighted that disclosure of genetic risk within families is not always straightforward. This supports clinical observations which suggest that some family members know of their risk, whilst others may not. This is important given that in the UK it is generally held that it is a family member’s responsibility to pass on this information. If genetic counsellors are to develop effective strategies to help patients deal with communication issues, we need to know more about what actually happens in families.

Methods - Interviews with people who had received genetic counselling for risk of Huntington’s disease (HD) and hereditary breast/ovarian cancer (HBOC), and some of their partners.

Results - In total, 37 consultants and 19 partners were interviewed. The analysis confirms clinicians’ observations that people tell some relatives but not others. Generic factors such as gender, kinship and family dynamics structured patterns of communication in both groups. However, there were notable differences between and within the groups.

Conclusion - The disclosure of genetic risk information within families at risk of both HD and HBOC can be a complex process. Some participants informed close and extended relatives whilst others were more prevaricative. The majority felt it was their role, not the doctors, to inform relatives of their risk. Clinicians responsible for the clinical care of several members of the same family, may find an understanding of patients’ individual approaches to disclosure might help understand why tensions exist and what counselling strategies are most likely to be effective.

ABSTRACT 014: Innovative Approaches to Improving the Acceptance of TextureModified DietsKaren Keast, Emma Power, Alison Anderson(NSW Huntington Disease Service, Westmead Hospital and Lottie Stewart Hospital Western Sydney Area Health Service - 40 Stewart St DUNDAS, NSW 2117 Australia)

The Dietitian and Speech Pathologists from the NSW Huntington Disease Service have looked at innovative approaches to enhance the acceptance of texture modified diets and to reduce the impact of such diets on people’s quality of life. In 1999, a resource package called Shop to Swallow was launched. It provided people with a range of foods available in the supermarket suitable to each of the soft, minced and puree textured diet categories. Photographs provided a visual display of foods to assist with client awareness and the food lists assisted people with planning and shopping. An evaluation sent to all purchasers, was extremely positive with 100% of respondents (n6) indicating that they were satisfied with the resource and that they had found it very useful. In 2002, a survey of popular fast food outlets and restaurant chains was conducted. A selection of foods were assessed and categorised into each of the soft, minced and puree diet textures. This resource will be compiled in a similar format to Shop to Swallow. Food molds alter the appearance of the pureed food to more closely resemble typically prepared food. A pre and post implementation study will be conducted in April 2003. The study will examine the perceptions of 12 adults with impaired swallowing and 12 adults with unimpaired swallowing to a range of pureed food items and to a range of molded food items. Results will be presented for the attribute ratings of overall liking; liking of taste, texture and appearance and ease of chewing and swallowing. This poster/paper will discuss the above innovative approaches for enhancing eating satisfaction and safety in both community and residential care settings. The implications of the findings and ideas for further research will be discussed.

We acknowledge the financial contribution of both The Australian Huntington Disease Association (NSW) and Lottie Stewart Hospital and the assistance from the Occupational Therapy and Food Service Departments at Lottie Stewart Hospital. There was no personal financial relationship with any products or sponsers.

ABSTRACT 023: Effects of rehabilitation on motor performance, mood state and everyday life of HD patientsPaola Zinzi (1), Graziano Graziani (2), Rosa De Grandis (2), Stefano Maceroni (2), Dario Salmaso(3), Annarita Bentivoglio (4), Marina Frontali (1) Paolo Zappata(2) Anna G. Jacopini(3)1) Institute of Neurobiology and Molecular Medicine, National Research Council, Rome, Italy, 2) Home Care;Nova Salus, Trasacco, Italy, 3) Institute of Cognitive Sciences and Technologies , National Research Council, Rome, Italy, 4) Institute of Neurology U.C.S.C., Rome, Italy

Rehabilitation therapy on Huntington’s Disease patients has rarely been object of a methodologically valuable attempt to assess its effects, data reported so far being mostly observational or anecdotal ones. We performed a research project aimed at obtaining reliable data on the effect of a multidisciplinary rehabilitation protocol for HD patients. The protocol, including motor, cognitive, speech and respiratory exercises, was based on a repeated 3-weeks admissions in the rehabilitation home. 42 HD patients were enrolled on the basis of the following criteria: disease stage I-III, a MMSE performance >16, absence of major psychiatric disorders. Here we report data on motor outcomes, effects on patients’ mood and on perception of whole experience by patients themselves and caregivers. A comparison of motor performance at each admission and discharge provides evidence of a significant improvement during the 3-weeks period showing the ability of patients to positively respond to treatment. The improvement however was not maintained till the subsequent admission (4 or more months later) and was partially or totally lost.

Additional data from patients and caregivers were collected through an ad hoc questionnaire exploring the effects of rehabilitation experience in everyday life: from the quality of family relationship and patient’s mood to his/her effective autonomy in performing the routine daily activities. A data preliminary analysis shows a beneficial effect on patient’s ability to communicate, interest in family and social life, motor control and engagement in activities. Repeated measures of depression levels on Zung Scale were also collected showing mild to remarkable improvements. The research was made possible by a grant from CNR to MF.

ABSTRACT 047: Improving Psychiatric Outcomes in an Outpatient HD PopulationTeresa L. Tempkin RNC, MSN, ANP, Ron Risley, MD, Vicki L. Wheelock, MD (University of California Davis Medical Center - Department of Neurology 4860 Y Street, Suite 3700 Sacramento CA 95817 USA)

Objective: To describe a care intervention to improve the availability and accessibility of mental health services in an outpatient HD population.Background: Mood and behavior changes are common problems in Huntington’s Disease. Problems can range from mild depression to psychosis. Access to mental health services is limited in most regions of the U.S., and particularly acute in the HD population because of a widespread belief that psychiatric symptoms in this population are ‘organic’ and therefore not amenable to traditional psychiatric treatment. Lack of available mental health care can lead to costly and painful involuntary psychiatric hospitalizations. This is especially true in the HD population where providers have limited experience with HD. Psychiatric symptoms are often successfully managed in the HD population, improving the quality of life for patients and their caregivers. To increase accessibility and availability of psychiatric services, a regional HDSA Center of Excellence (COE), in Northern California added a psychiatrist trained in HD to their outpatient treatment team. This provider manages the more complex psychiatric problems and is available to the HD triage team (nurse practitioner and social worker) when problems occur outside clinic time. Results: This COE has an average of 300 visits per year in its HD clinic. About 50% of visits are concomitant medical and psychiatric visits. Core clinic staff prior to July 2001 consisted of a movement disorder neurologist, an HD trained nurse practitioner, HD social worker and physical therapist. Prior to the addition of an HD trained psychiatrist in July 2001, 9 HD patients (3% of total HD visits) were involuntarily committed to inpatient psychiatric facilities. Since July 2001, there have been two (<1%) involuntary hospitalizations in the outpatient group the COE staff actively manages. The HD psychiatrist also evaluates each new referral and provides longitudinal follow up for existing patients.Conclusion: Access to specialized HD psychiatric services can improve management of psychiatric and behavioral problems in an outpatient HD setting measured as a decrease in involuntary psychiatric hospitalizations.

ABSTRACT 062: Parenting patterns in patients with Huntington’s Disease

Clement T Loy, Roslyn M Curran, Elizabeth A McCusker. (Huntington Disease Service, Westmead Hospital, Westmead. NSW 2145, Australia.

Background: Assisted reproductive techniques, such as pre-implantation genetic diagnosis, offer the potential to prevent transmission of Huntington’s Disease (HD) mutations. However, the decision to reproduce will also be affected by the future parenting ability of gene carriers, particularly if he or she develops manifest disease. We are not aware of any published data in the area, and have therefore carried out a pilot project studying the prevalence of parenting patterns and potential predictors amongst HD patients.

Methods: The HD Service includes 296 of the estimated 400 HD patients in the State of New South Wales. All patients in the Service with children aged 15 years or under, as of 1April 2003, were identified. Data were collected from a survey of health professionals and medical records. Statistical analyses were performed using SPSS 11.

Results: Thirty-four patients were identified, 2 excluded due to incomplete data. The median age was 42 years (26-60, 65% female). The median duration of disease was 5 years and median Independence Score 60 in 2003. Sixty-three children were identified, 2 excluded due to incomplete data. The median age was 10 years (1-15). 31% of the children were living with the affected parent. 21% of the children were mainly cared for by the affected parent. This was significantly more prevalent amongst children with affected mothers (31% vs. 8% for affected fathers, p=0.03). 28% of the children had no contact at all with the affected parent. Neither the timing of birth (before or after diagnosis of manifest HD), nor the gender of the child predicted the parenting pattern. Current Independent Scale scoring was also associated with the parenting patterns.

Conclusion: We have presented preliminary data on the parenting patterns amongst HD patients. A workup protocol for assisted reproduction should include considerations for this, and a neurological assessment.

ABSTRACT 066: Who killed that lady? A HD murder case in Japan

Kaori Muto, Ph.D., Department of Health Sciences, Shinshu University, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621 JAPAN, Yuki MiyaokaM Japanese Huntington’s Disease Network, 5-29-6 Minamisenju, Arakawa-ku, Tokyo, 116-0003 Japan, Takashi Nakajima, M.D.& Ph.D., Department of Neurology and Department of Clinical Research National Saigata Hospital, 468-1, Saigata, Ogata-machi, Nakakubiki-gun, Niigata, 949-3193, Japan

Objectives: A 72-year-old male caregiver murdered his 55-year-old wife with HD, by strangulation in 2002. His lawyer contacted with Japanese Huntington’s Disease Network and JHDN started to support him on the court. We conducted a study to clarify what caused him to kill her and place this case among all domestic murder cases in Japan.Methods: We examined written statements and interviewed with related persons including health care professionals and the accused.Results: He has cared for his wife at home for seven years since she started to show symptoms. First of all, we found many misunderstandings on HD and her physician gave some of them to him. For example, he was informed that people with HD would die within 6 or 7 years. He thought she was at terminal stage though she could live more. Second, he lost his hope to manage both work and caregiving. However, he didn’t make full use of social assistance for caregivers. He also tried to keep his reputation in his neighborhood as a good caregiver; who manages everything by himself. Finally, JHDN couldn’t access him directly though JHDN accessed the public health center that was in charge of care management of her disease.Conclusion: This case could have been prevented if her physicians and local health professionals had communicated with JHDN that had a role to educate them. The reason why is that it is not common that a lay organization to support local health professionals in Japan.We need to start a pilot model case for local health care network for HD to support people with HD and health professionals.

ABSTRACT 092: Self-Understanding and Identity: The Experience of Adolescents At Risk for Huntington’s Disease

Jessica L. Easton (University of British Columbia, Vancouver, British Columbia)

Adolescence is a time when individuals begin to explore and examine psychological characteristics of the self in order to discover who they really are and how they fit in the social world in which they live. It is during this time of self-exploration that adolescents at risk for Huntington’s Disease often learn of their risk status and witness the debilitating symptoms of the disease in their parents. Huntington Disease (HD) is an autosomal dominant neuropsychiatric disorder characterized by mid-life onset, involuntary movements, cognitive impairment, and depression.

This research investigated how adolescents experience living in a family with Huntington’s Disease and therefore at risk for Huntington’s Disease, and how this impacts their self-understanding and self-identity. The method of inquiry was based on a phenomenological approach. In-depth interviews were conducted with each of the adolescents, resulting in an extraction of three themes. These themes are: (1) Naming the Legacy: Understanding and Misunderstanding; (2) Experiencing the Legacy: Huntington’s Disease in Relation to Relationships; and (3) Integrating the Legacy: At the Crossroads of Self and Future Self.The analysis emphasizes that the at-risk adolescents’ exploration of self-identity and future self was an individual process influenced by the cognitive, developmental, and socio-cultural contexts of the adolescents’ lives. The process of learning about Huntington’s Disease occurred through intuition and practical and experiential learning. The adolescents found support outside their family through friends and adult mentors. They engaged in complicated coping strategies and demonstrated a capacity for decision-making that displayed maturity beyond what would be expected for their age group. These findings led to specific recommendations for theory, research, and clinical practice in the area of the adolescent experience of HD. The research underscores the need for healthcare professionals to re-evaluate their view of adolescent autonomy and capacity for decision-making.

TOPIC: PATHOGENETIC MECHANISMS

ABSTRACT 004: Homozygosity for CAG mutation in Huntington disease is associated with a more severe clinical course Ferdinando Squitieri, Milena Cannella, Vittorio Maglione (Neurogenetics Unit, IRCCS Neuromed, Localita’ Camerelle, Pozzilli (IS), Italy), Stefano Di Donato, Cinzia Gellera, Caterina Mariotti (Division of Biochemistry and Genetics, Neurological Institute IRCCS “C. Besta”, Milan, Italy), Giuliana Cislaghi (Division of Neurology, “Fornaroli” Hospital, Magenta (MI), Italy), David C. Rubinsztein (Cambridge Institute for Medical Research, Addenbrooke’ s Hospital, England), Michael R. Hayden, Elisabeth W. Almqvist (Centre for Molecular Medicine and Therapeutics, Dept. of Medical Genetics, University of British Columbia, Vancouver, B.C., Canada), David Turner (Dept. of Haematology and Genetic Pathology, School of Medicine, Flinders University, Adelaide, Australia) Anne-Catherine Bachoud-Lévi (Equipe Avenir, INSERM U421, Faculté de Medicine, Paris XII and Service de Neurologie, Hopital Henri Mondor, Creteil, France), Sheila A. Simpson (Department of Medical Genetics, Grampian University Hospitals Trust, Aberdeen, Scotland), Martin Delatycki (Victorian Clinical Genetics Services, The Murdoch Institute, Royal Children’s Hospital, Melbourne, Australia)

Huntington disease (HD) is caused by a dominantly transmitted CAG repeat expansion that is believed to confer a toxic gain-of-function on the mutant protein. In other poly(CAG) diseases such as the dominant ataxias, inheritance of two mutant alleles causes a phenotype more severe than in heterozygotes. In this multicenter study, we sought differences in the disease features between eight homozygotes and 75 heterozygotes for HD mutation. We identified subjects homozygous for the HD mutation by DNA testing and compared their clinical features (age at onset, symptom presentation, disease severity, and disease progression) with those of a group of heterozygotes, who were assessed longitudinally. The age at onset of symptoms in the homozygote cases was within the range expected for heterozygotes with the same CAG repeat lengths, whereas homozygotes had a more severe clinical course. The observation of a more rapid decline in motor, cognitive and behavioural symptoms in homozygotes was consistent with the extent of neurodegeneration as available at imaging in three patients and at the post-mortem neuropathological report in one case. Our analysis suggests that though homozygosity for the HD mutation does not lower the age at onset of symptoms it affects the phenotype and the rate of disease progression. More rapid worsening in homozygotes can be the consequence of a greater toxic effect due to the presence of two rather than one mutated protein products which may cause more aggregate formation and/or sequestration of critical cellular components in the targeted cells. An alternative explanation can be that CAG expansion homozygosity mimic a “recessive” loss of the neuroprotective wild-type huntingtin function, in addition to the dominant toxic effect of the mutation.These data, once confirmed in a larger series of patients, point to the possibility that the mechanisms underlying age at onset and disease progression in HD may differ.

ABSTRACT 017: Genetic disruption of tissue transglutaminase delays disease progression in a mouse model of Huntington’s disease

Craig D.C. Bailey and Gail V.W. Johnson (Department of Psychiatry and Behavioral Neurobiology, University of Alabama at Birmingham, 1720 7th Avenue South, Sparks Center, Room 1061, Birmingham, AL 35294-0017)

Tissue transglutaminase (tTG; TG2) catalyzes a calcium-dependent transamidation reaction of glutamine residues within specific protein substrates. Given the fact that tTG expression and enzymatic activity are elevated in Huntington’s disease (HD) brain, and that tTG can promote apoptotic events, it is important to determine whether tTG plays a role in the pathogenesis of HD. To test the hypothesis that tTG contributes to the pathogenesis of HD, a tTG knockout mouse line was crossbred with the R6/2 HD mouse model and the development and progression of the HD phenotype was examined. When compared with R6/2 mice that had a normal complement of tTG (R6/2; tTG +/+ mice), R6/2 mice with tTG knocked-out (R6/2; tTG-/- mice) exhibited a significant delay in the onset of motor dysfunction. Using the rotarod as a measure of motor function, R6/2; tTG -/- mice exhibited a greater latency to fall off the rotarod at 9, 10 and 11 weeks of age. However, tTG genotype had no effect on other R6/2 phenotypes such as the onset of tremor or characteristic clasping, and did not affect the weight loss that occurs in R6/2 mice. Interestingly, R6/2; tTG -/- mice exhibited a 20% delay in mortality compared with R6/2; tTG +/+ mice, suggesting that tTG ablation increases lifespan in the R6/2 mouse. These results demonstrate that tTG is involved in the pathogenesis of the HD phenotype in the R6/2 mouse and that tTG inhibition may ameliorate or delay disease progression in HD. (Supported by NIH grant AG12396. CDCB is the recipient of a postdoctoral fellowship from the Hereditary Disease Foundation. tTG knockout mice were a kind gift from Dr. Robert Graham.)

ABSTRACT 051: Analysis of huntingtin protein-protein interactions using the yeast two-hybrid system

Heike Goehler, Maciej Lalowski, Claudia Abraham, Bianca Bauer, Stephanie Waelter, Erich E. Wanker (Max Delbrück Center for Molecular Medicine, Robert-Roessle-Str. 10, 13125 Berlin, Germany) Renate Hasenbank, Uwe Worm, Hans Lehrach (Max Planck Institute for Molecular Genetics, Ihnestr. 73, 14195 Berlin, Germany)

Huntington’s disease (HD) is a progressive neurodegenerative disorder caused by an elongated polyglutamine tract in a large protein, huntingtin (htt), of unknown function. Several investigators have proposed that HD is caused by abnormal protein-protein interactions in neuronal cells. Therefore, we aim to identify interaction partners of wild type and mutant huntingtin in order to elucidate the cellular function of htt and to find potential therapeutic targets for HD. We use the yeast two-hybrid system for the identification of htt interacting proteins. In this study a set of 10 different htt fragments was screened against a human fetal brain cDNA library by cotransformation. Protein-protein interactions were identified by testing the growth of yeast clones on selective plates lacking histidine and uracil and by a beta-galactosidase filter assay. This approach resulted in the identification of 11 novel and two known huntingtin interaction partners. To verify the protein-protein interactions in vitro binding experiments, coimmunoprecipitations and colocalisation studies were performed.

ABSTRACT 087-LATE-BREAKING RESEARCH: Msh2 protein in base-excision mediated CAG expansion

Mechanism of trinucleotide expansion in Huntington’s Disease (HD) was shown previously to involve DNA strand break repair. In the present work we demonstrate directly in in vivo and in vitro models that breaks arise as a result of oxidative damage and when are repaired lead to expansion. DNA breaks are detected by comet assay in immature germ cells and in HD fibroblasts after treatment with hydrogen peroxide. We demonstrate that mismatch repair enzyme msh2 binds damaged HD locus after the treatment, possibly recognizing and prolonging lifetime of the formed hairpin. In contrast, DNA replication in both cell culture and animals in vivo does not result in expansion. Deletion of msh2 protein in these mammalian systems caused contraction at HD and microsatellite loci. Together our data support a mechanism in which expansion is a result of aberrant base excision repair of DNA damage caused by metabolic stress, where mismatch repair proteins play an important role stabilizing hairpin structures formed by the repeats.

TOPIC: CELL DYSFUNCTION

ABSTRACT 015: Specific progressive cAMP reduction implicates energy deficits in presymptomatic HD knock-in mice

Gines Silvia, Seong IS, Fossale E,Ivanova E, Gusella JF, Wheeler, VC, Persichetti F, MacDonald ME. (MGH, 149 building 13th street, Charlestown, MA 02129, USA) Trettel F (Department of Pharmacology, University of Rome La Sapienza, P.le A Moro 5, I00185 Rome, Italy)

Defects in gene transcription and mitochondrial function have been implicated in the dominant disease process that leads to the loss of striatal neurons in Huntington’s disease (HD). Here we have used precise genetic HD mouse and striatal cell models to investigate the hypothesis that decreased cAMP responsive element (CRE) mediated gene transcription may reflect impaired energy metabolism. We found that reduced CRE-signaling in HdhQ111 striatum, monitored by brain derived neurotrophic factor (BDNF) and phospho-CRE binding protein (CREB), predated inclusion formation. Furthermore, cAMP levels in HdhQ111 striatum declined from an early age (10 weeks), and cAMP was significantly decreased in HD postmortem brain and lymphoblastoid cells, attesting to a chronic deficit in man. Reduced CRE-signaling in cultured STHdhQ111 striatal cells was associated with cytosolic CREB binding protein (CBP) that mirrored diminished cAMP synthesis. Moreover, mutant cells exhibited mitochondrial respiratory chain impairment, evidenced by decreased ATP and ATP/ADP ratio, impaired MTT conversion and heightened sensitivity to 3-nitropropionic acid. Thus, our findings strongly suggest that impaired ATP synthesis and diminished cAMP levels amplify the early HD disease cascade by decreasing CRE-regulated gene transcription and altering energy dependent processes essential to neuronal cell survival.

ABSTRACT 039: Abnormal morphology of HD fibroblasts grown in enriched culture medium.

Lis Hasholt, Cathrine B. Petersen, Anne Nørremølle, Sven Asger Sørensen (Institute of Medical Genetics, The Panum Institute,University of Copenhagen)Klaus S. Frederiksen (Department of Molecular Genetics,NOVO Nordisk A/S, Denmark)

We have studied cultured fibroblasts from five HD patients with different CAG repeat length. All cultures showed a normal fibroblast growth pattern when the medium was supplemented with 10-15% fetal calf serum. However, fibroblasts from the HD patients lose their characteristic fibroblast like appearance when grown in a nutrient enriched culture medium. The cultures were instead composed of cells characterized by loss of processes, expanded cytoplasm and more intracellular space. The dysmorphology was most pronounced for fibroblasts obtained from a juvenile patient.The abnormal growth pattern of HD fibroblasts was reversible after reverting to medium with only 10-15% fetal calf serum. The addition of 20-mer phosphoro-thioated (PS-) oligonucleotides to the enriched culture medium also changed the growth pattern; the cells appeared like normal fibroblasts within a few days. This observation applied both to an antisense (PS-) oligonucleotide sequence and to control oligonucleotides, and is therefore not related to an antisense mediated down-regulation of huntingtin expression. PS-oligonucleotides binds non-sequence specific to many proteins, particularly to heparin-binding proteins including laminin and fibronectin. The oligo’s in our experiment may therefore interact with extracellular matrix proteins, which regulate cell attachment, migration and shape,and are also important in initiating cellular responses through classic signal-transduction pathways.The distribution of huntingtin is unchanged in the dysmorphic HD cells compared to normal control cultures. No inclusions were seen in HD fibroblasts. Preliminary results indicate that neither fibroblast growth factor (bFGF) nor insulin is key factors in development of dysmorphic cells. A detailed study of the morphological changes combined with investigations of altered gene expression profiles in these HD fibroblasts may contribute to clarification of the molecular pathogenesis in HD.This work was supported by grants from the Danish Medical research council, The Danish LHC Foundation and Sven Arend Larsen & Jon Johannesons Foundation.

ABSTRACT 058: A Quantitative Gene Expression Analysis Of Laser-Dissected Striatal Neurons In The R6/2 Mouse Model Of Huntington’s Disease

Birgit Zucker, David G Standaert, Anne B Young, Ruth Luthi-Carter, Sarah J Augood (Center for Aging, Genetics and Neurodegeneration, Massachusetts General Hospital and Harvard Medical School, Boston, USA)

Gene expression changes are observed in several animal models of Huntington’s Disease (HD), a neurodegenerative disorder caused by a CAG repeat expansion in the IT15 gene. In the striatum - a richly heterogeneous structure - projection neurons selectively die in HD whereas interneurons survive, and gliosis is present. We compared mRNA levels in striatal neurons harvested by Laser Capture Microdissection (LCM) in 12 week old R6/2 mice and wildtype littermates. Our samples comprised 50 dissected neurons which primarily represent projection neurons, the predominant type of neuron present within the striatum. Quantitative real time PCR revealed statistically significant differences in the neuronal expression of mRNAs encoding neurotransmitter receptors, neuropeptides and signaling molecules: dopamine-D2-receptor (39%), DARPP32 (35%), preproenkephalin (31%), hippocalcin (39%), complexin II (54%), all relative to wildtype. mRNA levels for the NR1 subunit of the NMDA-receptor did not change. Supporting an immune response during the disease process, b2-microglobulin, a mediator of cellular immune signaling, showed an increase (179% of control) in its mRNA level. The mRNA for the signal transducer and activator of transcription (STAT1) was also upregulated (194%) indicating changes in transcriptional regulation. This study shows that gene expression can be quantified in laser-dissected striatal perikarya in samples of as few as 50 mouse neurons. Our data demonstrate specific transcriptional changes in striatal neurons of R6/2 mice and suggest that these changes take place in neurons rather than in glia. We are now able to determine whether mRNAs are differentially regulated in other neuronal subtypes of the striatum and whether those potential differences contribute to the selective vulnerability of striatal neurons in HD.

Supported by the Hereditary Disease Foundation, the Deutsche Forschungsgemeinschaft and USPHS grant AG13617 (A.B.Y.). contact address: [email protected]

ABSTRACT 074: Heat shock protein 27 protects monkey Cos cells against oxidative stress generated by the expression of mutant huntingtin exon 1

Wance FIRDAUS, "Andreas WYTTENBACH", André Patrick ARRIGO

Huntington's diseases (HD) is one of ten known neurodegenerative diseases caused by expanded polyglutamine tracts. We show that overexpression of mutant huntingtin exon 1 in cells causes increased levels of oxygen species (ROS) that contribute to death. Our data suggests that the production of ROS is due to a dysfunction of mitochondria in cells containing huntingtin aggregates. Heat shock proteins Hsp 27 coexpression was found to reduce cell death by decreasing the intracellular ROS level. These data suggests that ROS directly contribute to cell death in polyglutamine disease and highlights the importance of Hsp27 as an antagonist of this process.

mailto:[email protected]

ABSTRACT 075: Huntingtin expression results in a decrease of proteasome activity and triggers apoptosis in pc12 cells.

George M. Lawless, Allan J. Tobin, Erik S. Schweitzer (University of California Los Angeles, 695 Charles Young Dr. Los Angeles CA 90095)

We have developed stable PC12 (a rat pheochromocytoma cell line with neuronal properties) cell lines that express exon-1 of the huntingtin gene, encoding polyglutamine of either 25 or 103 repeats, fused to the reporter, EGFP (HttQ25-EGFP or Htt Q103-EGFP). Expression of HttQ103-EGFP results in the formation of aggregates and is toxic to the cells whereas, expression of HttQ25-EGFP does not produce aggregates and is not toxic. It appears that the ubquitin-proteasome system is unable to properly degrade the misfolded expanded repeat (Q103) Htt protein; resulting in the formation of aggregates. We have found that expression of HttQ103-EGFP decreases proteasome activity while expression of HttQ25-EGFP does not. Furthermore, we have found that Caspase-3 is activated in PC12 cells expressing Htt Q103-EGFP. We propose that the toxicity of HttQ103-EGFP in PC12 cells is apoptotic. Supported by grants from NINDS and CURE-HD Initiative of the HDF.

TOPIC: OTHER

ABSTRACT 002: Huntington disease in Cuba.

Dra. Tatiana Zaldívar Vaillant*, Dr. Enrique Mitchel Esteban*, Dr.Esther Alonso*, Dr. Gloria Lara*, Dr: Alejandro Negrín**, Dr. Claudio Scherlie**, Ada Beatriz Portal*, *National Neurology and Neurosurgery Institute, ** Hermanos Ameijeiras Hospital

Even when in Cuba we don’t have the concept of association families in several diseases, we have created an interdisciplinary group, in order to support HD families in our country. The composition of this group is as follow: 4 Neurologist, 1 Clinical Genetics MD, 1 Psychologist, 1 molecular Biologist, 1 Psychiatrist and 1 Social worker.

After we organised our methodology in the organisation of our HD Clinic, started working in this way (last june 2002). One day per month (each Friday fourth at 1:00PM) our group have a meet with no more than three families. Usually comes a patient with a relative. Until now we have registered 10 families.

RECOMMENDATION AND PROJECTION FOR FUTURE

All this ideas have arisen since the development of our work, interdisciplinary discussion, theoretical updating, and advice we have been soliciting to colleagues with experience in this subject in other countries.

To develop a specialized psychiatric attention to HD patients. Establish this clinic continue as a work methodology for all HD families in our country. Molecular diagnosis: Start this when we could get the chemical reagents. While we have not had the possibility for molecular diagnosis, we can not start working in predictive tests for relatives in risk. Any way this would be a future step, so in this period we are accumulating experience in the ethical procedures these kind of tests require. Our aim and intention is to reach a consolidation and a strong experience as a group, in order to organize a kind of meeting, with patients and relatives, to inform them the updating information in advances of treatment and research of the disease.

ABSTRACT 007: The effect of Huntington’s disease on the ability to respond to conflicting spatial stimuli

Dr Nellie Georgiou-Karistianis, Prof. John l. Bradshaw (Experimental Neuropsychology Research Unit, Psychology Department, Monash University, Clayton, Victoria, Australia), Prof Edmond Chiu (Huntington’s Disease Clinic, Melbourne University, Melbourne, Victoria, Australia), Dr Andrew Churchyard (Monash Neurology, Monash Medical Centre, Clayton, Victoria, Australia)

This study examined the efficiency with which Huntington’s disease (HD) participants could shift and direct attention away from conflicting stimulus response configurations.

Various relevant (congruent) and irrelevant (incongruent) spatial stimuli were adopted from a paradigm developed from the Simon effect. Stimuli consisted of left and right pointing arrows, and in some cases, conditionality manipulations were also employed (i.e., “=” or “x”) that instructed participants to either maintain or change cognitive set, respectively. As predicted, HD participants, regardless of medication status and unlike controls, were particularly disadvantaged in responding to various conflicting stimulus response configurations. HD participants may experience difficulties with attentional shifts and in changing cognitive set; they may also be more susceptible (than controls) to the conflict that can arise when the spatial code formed for the stimulus is irrelevant for selecting the appropriate response. We conclude that our findings support the notion that cognitive deficits in HD may stem from abnormalities of the major pathways interconnecting the basal ganglia and the frontal lobes.

ABSTRACT 011: Small compounds that inhibit mutant Hungtingtin aggregation: Potential therapeutics for HD patients

Jin Wang, Syvia Gines, Marcy E. MacDonald, James F. Gusella (Molecular Neurogenetics Unit, Massachussetts General Hospital, Harvard Medical School, 149 13th Street, Charlestown MA, 02129, U.S.A.)

Considerable evidence supports the view that neurodegeneration in Huntingtin’s disease (HD) is triggered by an expanded polyglutamine segment that alters the conformation of mutant huntingtin Relative to its normal counterpart. The hypothesis that a conformational property capable of promoting aggregation is the crucial element in triggering HD pathogenesis leads directly to the notion that compounds that affect this property could have therapeutic potential. At present, it is uncertain whether the initiator of pathogenesis involves an insoluble form of huntingtin’s amino terminus or an altered interaction of a soluble form of huntingtin. The identification of compounds that either inhibit aggregation or promote it by altering the soluble polyglutamine conformation would provide valuable research tools to test this question in model systems and could help guide future development of human therapeutics for HD and the other polyglutamine disorders. Consequently, we used an in vitro aggregation assay to screen the NINDS Custom Collection of 1040 FDA approved drugs and natural products. This assay involves in vitro aggregation, using thrombin digestion of GST-Htt(1-171)–Q58 in a standard experimental paradigm, where 100% of the Htt-Q58 moiety can be detected in amyloid-like aggregates after 24 hours incubation at room temperature. From the 1040 small compounds, we identified 12 compounds that inhibit huntingtin aggregation in vitro at low EC50 (<20uM) and 5 compounds that promote aggregation. We currently are evaluating these interesting compounds in a mutant huntingtin neuronal progenitor cell-based assay to gain clues to their potential effectiveness in vivo. Preliminary evidence indicates that at least some of the drugs restore the mutant cell phenotype to wild type, suggesting that they are candidates for testing in mouse models of HD.

ABSTRACT 013: An abnormal sleep pattern and excessive movements can characterize patients with Huntington’s diseaseMarieta H. Anca Department of Neurology,Wolfson Medical Center,Sakler Faculty of Medicine ) Nir Giladi, Amos D.Korczyn (Department of Neurology, Tel Aviv Medical Center, Sackler Faculty of medicine),A. Blonder (Maarag- Worldclass Sleep Diagnostic Center) Tel Aviv, Israel

Background: Huntington’s disease (HD) is an inherited neurodegenerative disease characterized by motor, cognitive and psychiatric symptoms. Over 80% of patients complain about sleep disturbances. There are evidences that GABA plays a major role in sleep regulation. The sleep studies done on HD patients were non-conclusive. We hypothesize that sleep disturbances are early and prognostic signs of the disease.Objective: To assess the sleep pattern in genetically tested HD patients with mild to moderate disease and to correlate it with clinical features.Methods: Ten HD patients (7 female), with a mean disease duration of 6.5 ± 3 years, underwent one night polysomnographic (PSG) assessment. All patients had motor, mood and sleep disturbances and complained of low daily performances. 6 patients are treated with neuroleptic drugs. All patients were assessed for their motor, mood and cognition scores using the Unified HD Rating Scale, the Hamilton questionnaire and the Mini-Mental State Examination (MMSE).The sleep quality was assessed using the Epworth Sleep Scale The PSG was performed at patient’s home , in order to prevent the “first night effect” of sleep laboratory. The Embla system digital recorder of 16 channels was used . The sleep scoring was done following the Rechtschaffen and Kales protocol.Results: The study population was characterized by a mean age of 51.5+1 years, a mean score for Functional Capacity of 9.1±2 , for MMSE of 25.7±5.6, for Hamilton test of 18.3±8.9, for Epworth scale of 35.4±11.7. All patients completed a whole night PSG. The sleep architecture, compared to standard sleep in normal age-matched subjects revealed prolonged NREM1 and reduced amount of REM (p< 0.001). Multiple body movements in sleep were recorded in all patients, mainly during NREM sleep.Conclusion: The HD patients has an abnormal sleep pattern. The excessive movements does not explain the poor sleep. This pattern seems to be an intrinsic feature of the disease not modifiable by disease duration or neurolepic treatment.

ABSTRACT 025: Comparisons of CAG repeat distributions for familial and sporadic HD cases & intermediate allele frequencies for different populations support a stepwise model for CAG expansion.Lilias H Barron, Mary E Porteous, Jonathan P Warner (Clinical Genetics Service, Molecular Medicine Centre, Western General Hospital, Edinburgh EH4 2XU, UK ), Brian Smith (Medical Genetics, Duncan Guthrie Institute, Yorkhill NHS Trust, Glasgow G3 8SJ, UK), Sheila A Simpson (Medical Genetics, Aberdeen Royal Infirmary, Foresterhill, Aberdeen AB9 2DZ, UK), David R Goudie (Human Genetics, Ninewells Hospital, Dundee DD1 9SY, UK) Steven Evans (Regional Genetics Service, Sheffield Children’s Hospital, Sheffield, UK)

We provide a national diagnostic service for Huntington’s disease (HD) in Scotland. Since 1993 when huntingtin was identified we have carried out over 1000 CAG mutation tests. From our HD family analyses we present CAG repeat size determinations seen in parent to child transmissions for over 200 HD meioses and also over 400 non-HD meioses. Paternal transmission gives rise to a significantly greater number of size changes than does maternal. The majority of changes from an affected parent involve a gain or loss of one repeat or no change:54%of paternal, 77% of maternal. These transmission data suggest that the CAG repeats are considerably more stable than generally suggested in the scientific literature. We present a comparison of CAG repeat size distributions in a cohort of sporadic cases versus affected individuals with a known family history. A Mann Whitney U test shows an over representation of the 36-41 CAG repeat range in the HD patients with no previous family history (p<0.0001). This supports a stepwise model of mutational expansion as the main source of HD in our population. The CAG repeat size range between 27 and 35 repeats represents an unusually high figure of >3% of normal CAG alleles in Scotland. Differences in the frequencies of alleles in this “feeder” range are seen when comparisons of frequencies for populations from the East and West coasts of Scotland, Yorkshire and published world data are made. The

significance of these observations and the possible influence of “feeder” allele frequencies on prevalence of HD are discussed.

ABSTRACT 027: Multidisciplinary Care in the Information Age a system for coordination of care amongst geographically dispersed providers

Ronald P Risley MD, Teresa L Tempkin RNC MSN ANP (University of California, Davis)

A multidisciplinary approach to care of people with Huntington’s Disease has been shown to improve quality of life. A true multidisciplinary approach, however, requires close coordination among the participants. Working with rare disorders presents particular challenges to the multidisciplinary team: even when specialists are concentrated (as in the HDSA Centers of Excellence) the patient volume is often not sufficient to justify full-time specialist positions. Often specialty care must be coordinated among providers who must also work at geographically dispersed sites. We recognized the need for an electronic communication system when we observed that electronic mail (email) was playing an increasing role in coordination of activities at the nascent HDSA Center of Excellence at UC Davis (the Center). Email served administrative needs well, offering accessibility, ubiquity, timeliness, and accountability. Email was inappropriate for specific patient care issues, however, because the public Internet email system cannot assure the privacy of patient-related communications. Secure email solutions exist, but these generally require custom software and configuration at each workstation. Given the diversity of sites where our providers operate (a University hospital, county clinics, private clinics, and administrative sites), solutions which required modification of workstations in any were deemed impractical. We describe a novel approach to coordination of a multidisciplinary team that leverages ubiquitous technologies originally developed for electronic commerce, such that any Internet-connected workstation with a standard web browser can access Center communications while still assuring patient confidentiality. The system as implemented at the Center allows the treatment team to communicate rapidly and unobtrusively from a diverse range of institutions, sites, and practice settings while preserving patients’ privacy and avoiding the need to modify other institutions’ equipment. In addition, the system was realized without a significant investment in infrastructure or training, and is being maintained at virtually no cost to the Center.

ABSTRACT 029: The huntingtin-interacting protein endophilin A3 forms filamentous structures which associate with microtubules but not with actin filaments

Alis. C. Hughes, Rachel. J. Errington, A. Lesley. Jones. (UWCM, Heath Park, Heath, Cardiff.)

Endophilin A3 is a member of the endophilin family of proteins, thought to play a role in the formation of clathrin-coated vesicles from the plasma membrane. This protein may have a role in Huntington’s disease pathology as it associates with huntingtin in a repeat-length dependent manner and is expressed at its highest levels in the brain. We investigated the localisation of endophilin A3 in mammalian cells expressing the endophilin A3 gene fused to green or cyan fluorescent proteins (GFP, CFP). Endophilin A3 did not localise around the periphery of the cells as was expected but formed numerous bright puncta and filamentous strands, superimposed on top of a diffuse background. Colocalisation studies revealed the endophilin A3 structures to be distinct from mitochondria, endoplasmic reticulum, lysosomes and actin, but to associate with subsets of microtubules. Perturbation studies with nocodazole, colcemid (microtubule disruptors) and cytochalasin D (actin filament disruptor) confirmed these findings. We therefore propose that endophilin A3 has a role in transport along or in the structure of microtubules, in addition to its role in endocytosis. We are now investigating the dynaminc colocalisation of full-length huntingtin fused to YFP with normal or expanded polyglutamine and endophilin A3 in both neuronal and non-neuronal cells.

ABSTRACT 040: High throughput screen to identify compounds that downregulate intracellular mutant huntingtin expression

Russel DE*, Maxwell MM*, Fox J*, Thompson LM**, Hersch SM*, Housman DE***, Young AB*, Kazantsev A*(*Center for Aging, Genetics and Neurodegeneration, Massachusetts General Hospital, **Department of Biochemistry, University of California at Irvine, ***Center for Cancer Research, Massachusetts Institute of Technology)

150,000 people nationwide are at risk of developing Huntington’s Disease (HD), which at onset, initiates a clinical phenotype that progressively worsens. In an inducible transgenic HD mouse model it was shown that when mutant huntingtin (htt) protein expression is blocked, the progression of the HD neurological phenotype is arrested. Furthermore, when soluble and aggregated forms of mutant huntingtin within disease-affected neurons are cleared, the neuropathological symptoms associated with HD are reversed. We have designed a high throughput screen to identify small chemical agents that assist cellular degradation machinery in eliminating mutant huntingtin accumulation.

As published, we created a PC12-based stable cell line, which expresses a mutant (104Q) huntingtin-GFP fusion protein when induced. From data obtained using expanded polyQ-specific antibodies, we determined that in the absence of expressed protein, mutant huntingtin polypeptides are quickly degraded. The degradation of huntingtin-GFP polypeptide is directly correlated with decreasing fluorescent signal. Our screen is based on this premise. We conducted a preliminary 3,000 compound screen to validate our assay. Currently we are in the process of screening our complete chemical library totaling 30,000 compounds. We expect our cell-based screen to yield compounds that reduce the level of mutant huntingtin expression via transcriptional regulation. We will discuss the structural and functional relationship of “hit” compounds to the degradation rate of pathological and abnormally folded conformation of mutant huntingtin. Ultimately, our purpose is to analyze the efficacy of compounds identified in the screen in HD animal models.

This work is supported by the Center for Aging, Genetics and Neurodegeneration (CAGN).

ABSTRACT 045: PEG feeding in end stage Huntington’s disease: an examination of the clinical and ethical issues, including the views of the doctor, patient and the patient’s family.

Sheila A Simpson, Kofi Atiemo (Grampian University Hospitals Trust, Aberdeen, Scotland, UK)

Clinical features of Huntington’s Disease (HD) include movement disorder, personality changes and cognitive impairment. Classically deterioration progresses until excessive weight loss and uncoordinated swallow with risk of aspiration occurs. Feeding using percutaneous endoscopic gastrostomy (PEG) has been suggested as a means of providing additional calories in a safer and more reliable fashion.PEG feeding raises a number of ethical issues because it is a form of artificial feeding and it does not remove the risk of aspiration. It will not cure HD, but it may sustain life. None the less it is often put in place with little or no discussion with the patient or his family. There has been little debate about withholding or withdrawal of such feeding which some view as treatment and others as basic care, yet the dilemma occurs frequently as the HD patient deteriorates. We surveyed the views of HD patients, relatives, carers and healthcare professionals in Scotland regarding a patient’s right to choice with regard to PEG feeding in the end stage of HD. The minority of doctors encouraged patients to discuss issues of end life care and very few doctors sought their patient’s opinion regarding PEG feeding early in the natural history of HD despite 93% of respondents stating that the patient should be allowed to make the decision. We examine the evidence for and against the use of PEG feeding in this paper. We suggest that early in the course of HD clinicians should discuss with the patient and their carer the prospect of PEG feeding, since at that time patients are competent and able to communicate their wishes. Their views should be

recorded. Respondents agreed that guidelines on how this could be achieved would be acceptable to patients and healthcare professionals.

ABSTRACT 052: Motor deficits and striosome volume in the YAC72 mouse model of Huntington’s disease.Collene Lawhorn, Diane M. Smith, Lucy L. Brown (Albert EInstein College of Medicine, Dept of Neuroscience/Neurology, Bronx, NY 10461) Blair R. Leavitt (University of British Columbia, Vancouver Canada)

The striatum is a heterogeneous structure with distinct neurochemical and functional compartments called striosomes, which are embedded in an extrastriosomal matrix. In early-stage Huntington’s disease (HD), there is some evidence that cell loss is specific to striosomal compartments (Hedreen and Folstein, 1995). To investigate whether striosome pathology mediates some of the motor symptoms in HD we used a yeast artificial chromosome transgenic mouse model of HD that expresses full-length human huntingtin with 72 polyglutamine repeats (YAC72), because it showed neuronal degeneration at 12 months and behavioral changes starting at 7 months of age (Hodgson et al, 1999). We ran 15-month old YAC72 mice (n) and age-matched controls (n) on assays for motor function. Prior to testing, all subjects were blindly screened for general health using the complete SHIRPA protocol. Compared to controls, mutant mice showed a 70% decrease in their ability to remain on a suspen! ded wire in the wire maneuver task (Controls: 14±3 seconds; Mutants: 4±2 seconds). Footprint analyses revealed a 16% increase in the width between paws during locomotion compared to controls (Controls: 18±0.7 mm; Mutants:21±1.2 mm). We used 3D reconstructions of coronal brain sections to determine striosome volume. Overall striatal volume did not differ between YAC72 and control mice. However, striosome volume was 29% less than controls in the YAC72 mouse throughout the striatum, especially in posterior regions (Controls: 14±1 percent of striosomes relative to striatum; Mutants: 10±1 percent of striosomes relative to striatum). Most importantly, wire maneuver scores were positively correlated with striosomal volume, especially in posterior regions, yielding correlation coefficients of .76 across groups and .90 for controls alone. These data are the first to show a striosomal volume loss in a transgenic mouse model for Huntington’s disease, and the first to demonstrate a co! rrelative relationship between natural motor behaviors and any strioso mal measure.

ABSTRACT 076: Bilateral GP stimulation for Huntington’s DiseaseElena Moro, Yu Yan W. Poon, Serena Hung, Anthony E. Lang (Movement Disorders Center, University Health Network, Toronto Western Hospital, Toronto, On, Canada), Pablo M. Arango, Andres M. Lozano (Division of Neurosurgey, University Health Network, Toronto Western Hospital, Toronto, On, Canada)

Background: Huntington’s disease is an inherited progressive neurodegenerative disorder clinically characterized by involuntary choreiform movements, cognitive impairment and psychiatric symptoms. The severe neuronal loss in the striatum leads to overactivity of GPe, underactivity of STN, GPi and thus overactivity of the thalamus, resulting in chorea. GP DBS is a relatively new functional surgery which has been proven effective in PD, dystonia and other movement disorders.Objective: to determine if GP stimulation is effective in improving chorea in HD.Methods: A 42-year-old HD gentleman was selected for bilateral GP stimulation because of the severity of his chorea and his relatively preserved neuropsychological and psychiatric conditions. Before surgery he also had severe speech, swallowing, balance and gait impairment. His UHDRS motor assessment score was 86. In particular, his dystonia total score was 16, whereas the chorea total score was 25. Bilateral GP DBS was performed using Leksell’s frame and MRI imaging under local anesthesia. The positioning of the electrodes (Medtronic 3387) was determined using microrecording and micro- and macro-stimulation. A Kinetra IPG was implanted in the right subclavicular area one week after.Results: Immediately post-op, a moderate pallidotomy-like effect was observed and lasted for one week. Thereafter, several settings of the electrical parameters of stimulation were assessed at each contact over 6 weeks. The best clinical result was obtained with contact 2 bilaterally. High frequency stimulation (130 Hz) worsened bradykinesia, speech and balance, whereas low frequency stimulation (40 Hz) improved chorea without motor side effects. His 2-month post-operative UHDRS motor assessment score was 64 (-26%), his dystonia total score was 10 (-37.5%) and chorea total score was 11 (-56%). In addition, speech, swallowing and gait showed a moderate improvement. Conclusion: bilateral GP stimulation appears to be a promising treatment for chorea and dystonia in HD.No personal financial relationship with any products or sponsors.

POSTER ABSTRACT SESSION #2MONDAY, AUGUST 18, 2003

8:00 a.m. – 9:00 a.m.

All Congress posters will be on display Sunday and Monday.

We request poster presenters to stand by their poster during poster abstract session #2 on

Monday, August 18 8:00 a.m. to 9:00 a.m.

The following poster topics have been assigned to poster session #2:

Neuronal Vulnerability and Survival,Cellular-Based and Genetic Animal Models, Animal Model Experimental Therapeutics,

Clinical Characteristics, Neuroimaging,

Human Experimental Therapeutics

(Abstracts for the above topics to follow…)

TOPIC: NEURONAL VULNERABILITY AND SURVIVAL

ABSTRACT 061: mRNA Profiling of Two Striatal Neuron Populations with Differential Vulnerability in HD: A Study Utilizing LCM, Real-Time PCR, and cDNA Microarrays

Jordan D. Scopa, Christine E. Keller-McGandy, Xiaojia Ren, David G. Standaert and Sarah J. Augood.

The neuroanatomical hallmark of Huntington’s Disease (HD) is severe atrophy of the striatum. Medium-sized interneurons expressing neuronal nitric oxide synthase (nNOS)/somatostatin (SOM) are spared, but there is selective loss of the medium spiny projection neurons. These different cell types are intermingled throughout the striatum and can be identified using immunostaining, and harvested using laser-capture microdissection (LCM). Differential cDNA microarray profiling has already been carried out in the laboratory on these populations of NOS/SOM striatal interneurons versus medium-sized projection neurons in the control adult rat in order to identify differences in mRNA profiles that may account for their differential vulnerability. Quantitative analysis of the data identified 10 cDNAs that are differentially expressed in the two cell populations. The differential expression of the mRNAs encoding VL30, a transcriptional element, and phospholipase C delta 4 (PLCd4), a generator of inositol 1,4,5-trisphosphate and diacylglycerol, are of particular interest, as they play a role in Ca2+ and receptor-mediated signaling. These data suggest that dysregulation of calcium signaling may contribute to the differential vulnerability of these two striatal cell types in HD. These findings are currently being validated utilizing real-time PCR and investigated further in the R6/2 transgenic mouse model of HD.

Supported by the Hereditary Disease Foundation (S.J.A.), the Huntington Disease Society of America (S.J.A.), and Harvard College Research Program (J.D.S.)

TOPIC: CELLULAR BASED AND GENETIC ANIMAL MODELS

ABSTRACT 037: Interaction between huntingtin and the nuclear receptor corepressor

Nagehan Ersoy, A. Nazli Basak Bogazici University, Department of Molecular Biology and Genetics, Istanbul, Turkey, Lesley Jones University of Wales, College of Medicine, Institute of Medical Genetics, Cardiff, UK

Transcriptional dysregulation is one of the proposed pathogenic mechanisms by which mutant huntingtin (htt) exerts its effects. Abnormal interactions of the mutant htt, with the proteins involved in transcription, may lead to transcriptional dysregulation. Boutell et al (1999) detected a repeat-length-dependent interaction of htt with the nuclear receptor corepressor (NCoR). NCoR mediates repression from sequence specific nuclear receptors, and its level in cells is thought to be a limiting factor in mediating repression. We proposed that mutant htt can sequester NCoR from its normal site of action and thus alter transcription. Full length (FL) htt and NCoR fusion proteins were generated with living colors vectors (Clontech, USA). FL htt with 25, 60 or 90 glutamines(Q) was cloned into EYFP; FL NCoR was cloned into ECFP and EGFP. HEK 293 and PC12 cells were transfected with the fluorescent constructs and analyzed with laser scanning confocal microscopy. Normal and mutant htt fusion proteins showed diffuse expression in the cytoplasm. Cells transfected with htt 25Q constructs were cytologically normal, however, cells transfected with htt 60Q and 90Q developed aggregates and presented with apoptotic morphology. NCoR fusion proteins exhibited punctate expression in the cell nucleus and were excluded from nucleoli. Co-transfection studies revealed that NCoR localization in the cell was apparently not altered when expressed together with mutant htt. Mutant htt fragmentation was detected on Western blots; characterization of the fragments is in progress. Further studies include cloning of DsRED into YFP-tagged htt, to visualize the cellular compartmentalization of the htt fragments, and detailed localization of NCoR by immunocytochemistry and confocal microscopy.No personal financial relationship.

ABSTRACT 042: Identification in Cell Culture of Small, Recruitment Positive Polyglutamine Aggregates: Aggregation FociErica Johnson (UT-ORNL, Genome Science and Technology Program, University of Tennessee, Knoxville, TN 37996), Valerie Berthelier, Alexander Osmand, Ronald Wetzel (Graduate School of Medicine, UT Medical Center, Knoxville, TN 37920)

The possible role of various aggregated states of polyglutamine (polyGln) in the disease mechanisms of expanded CAG repeat diseases continues to be a subject of great interest. We believe that some studies probing the relationship between aggregate accumulation and cell death or dystrophy may be compromised by an inability to visualize and/or quantify the most important, most toxic, aggregated species. For example, small polyGln aggregates in vitro are much more potent on a weight basis than are large aggregates in the recruitment, through a continuing elongation reaction, of other polyGln sequences [1]. This suggests that measurements that focus only on very large aggregates, like neuronal intranuclear inclusions, may entirely miss the aggregates that are most important for cytotoxicity via recruitment of other polyGln proteins. We have developed methods that allow imaging of relatively small species (presumably aggregated polyGln proteins) in archival Huntington’s disease brain sections based on their ability to recruit new polyGln peptides, species we refer to as aggregation foci (AF). (A. Osmand, V. Berthelier and R. Wetzel, manuscript in preparation). We present here an extension of the same basic approach to the detection and quantitation of AF in cell culture. PC-12 cells as well as SH-SY5Y neuroblastomas were transiently transfected with constructs encoding expression of green fluorescent protein fused to huntingtin Exon 1 fragments including various polyGln repeat lengths, both benign (Q25) and pathological (Q47, Q65, and Q103). After fixation, the cells were incubated with nM concentrations of biotinylated-polyGln Q28 monomer followed by incubation with fluorescently tagged streptavidin-conjugate. Fluorescent foci indicative of the presence of recruitment-competent aggregates were only observed in cells transfected with the pathological length exon 1 constructs. In addition, these polyGln recruitment-competent aggregates appeared prior to the appearance of mature polyGln aggreg ates (detected as green fluorescent foci), indicating that at this early stage their structures are too small to be detected by the GFP signal. These structures are active in recruiting polyGln sequences in contrast to the larger polyGln aggregates detected by green fluorescence, which appear to give weaker recruitment responses. The ability to independently measure both mass and recruitment activity should allow closer look at the aggregation process in the cell and thereby offer new insights into the role of aggregation in cell death. This research funded by the Hereditary Disease Foundation. Reference: 1. Chen, S., et al., Polyglutamine aggregation behavior in vitro supports a recruitment mechanism of cytotoxicity. J Mol Biol, 2001. 311(1): p. 173-82.

ABSTRACT 054: Toxicity of a carboxyl-terminal huntingtin fragment in DrosophilaHui Zhu, Hiroki Takano, Jo-Chen Chou, Naoto Ito, James F. Gusella Molecular Neurogenetics Unit Massachusetts General Hospital/Harvard Medical School BLDG 149, 13th St. Charlestown, MA 02129

Cleavage of both wild type and mutant huntingtin has been demonstrated in vitro and in vivo and considerable attention has been paid to the possibility of toxicity due to the amino terminal polyglutamine-containing fragment. We have noted abnormal phenotypes in transgenic flies that overexpress a carboxyl terminal fragment of Drosophila huntingtin. Driving ectopic expression of the fragment in the fly nervous system with ELAV produced significant defects in climbing ability late in aged flies (7 weeks old). Overexpression in fly eyes, driven by GMR, caused obvious roughness and pigmentation loss. Semi-thin sections of the affected eyes showed disrupted ommatidia and large vacuoles, suggesting extensive cell loss. Electron microscopic studies indicated loss of photoreceptor neurons within the ommatidia. Co-expression of viral caspase inhibitor, p35, with the huntingtin fragment significantly rescued the eye phenotype, supporting the view that the C-terminal huntingtin fragment caused apoptotic cell death. These data raise the possibility that generation of a stable amino-terminal huntingtin fragment could produce deleterious consequences in HD brain and participate in HD pathogenesis. We are currently carrying out a genetic modifier screen using the eye phenotype in these transgenic flies and several candidate modifiers genes have been identified. Validation of these modifiers and assessment of their roles in the normal function of huntingtin and potentially in modifying disease progression will require further work. We hope that these investigations will broaden our understanding of HD pathogenesis and identify potential leads to the treatment of HD by modifying disease progression.

ABSTRACT 064: Suppression of polyglutamine toxicity by a Drosophila ortholog of MRJ

Zahra Fayazi, Susan Marion, Marlene Shero, Xiankun Bao, Christopher Hand, and Parsa Kazemi-Esfarjani (Department of Physiology and Biophysics, Center for Neuroscience, School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, New York, USA 14214)

We have shown that Drosophila fruit flies that express proteins with expanded polyglutamine repeats in the retina, beginning at the time of retinal cell differentiation, have eyes with little or no pigmentation and structural integrity. To screen for genetic modifiers of polyglutamine toxicity, these flies were crossed to those with random chromosomal insertions of a transposable P-element, containing a strong recombinant transcription enhancer element. If a P-element were inserted near or within a modifier locus, in the progeny, the insertion alone or the enhancer within the P-element would modulate the transcription of the gene(s) in that locus. Thus, the eye phenotype was either enhanced or suppressed. One such P-element that partially suppressed the loss of eye pigmentation, was inserted 9.7 kb upstream of a gene that encodes an ortholog of a brain-enriched chaperone, the human MRJ (mammalian relative of DnaJ). In transfected 293 cells, MRJ reduced and delayed the aggregation and cellular toxicity of a mutant exon 1 huntingtin, with 150 glutamine repeats (1). To investigate the effect of the Drosophila ortholog (dmrj) on the aggregation and toxicity of expanded polyglutamines, four different transgenic P-element constructs were produced, expressing full-length dmrj cDNA at different levels. A fifth construct carried a dmrj ORF with an N-terminal Flag epitope tag. For each construct, at least three lines of transgenic flies were established. The progeny from the cross between dmrj transgenic flies and those expressing an expanded polyglutamine showed that, similar to its mammalian counterpart, dmrj suppresses the toxicity of expanded polyglutamines. We are now examining its effect on the aggregation of polyglutamines.References:1. Chuang JZ, Zhou H, Zhu M, Li SH, Li XJ, Sung CH (2002) Characterization of a Brain-enriched Chaperone, MRJ, That Inhibits Huntingtin Aggregation and Toxicity Independently. J Biol Chem 277:19831-8.

ABSTRACT 077: The role of calpain in Huntington’s disease

Lisa M. Ellerby, Juliette Gafni, Evan Hermel Anna Logvinova(Buck Institute for Age Research, Novato, CA 94945) Roger Siman (Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104) Cheryl L. Wellington, Michael R. Hayden (Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia V5Z 4H4 Canada; Center for Molecular Medicine and Therapeutics, Department of Medical Genetics, University of British Columbia, Vancouver, British Columbia, V5Z 4H4, Canada)

Huntington’s disease (HD) is a neurodegenerative disorder caused by a glutamine expansion near the N-terminus of huntingtin (Htt). Abnormal proteolytic processing of mutant Htt may play a critical role in disease progression and pathogenesis. Recent work indicates calpains may participate in the increased and/or altered patterns of Htt proteolysis leading to the selective toxicity observed in HD striatum. Here, we identify two calpain cleavage sites in Htt and show that mutation of these sites renders mutant Htt less susceptible to proteolysis and aggregation, resulting in decreased toxicity in an in vitro cell culture model. In addition, we found calpain- and caspase-derived Htt fragments preferentially accumulate in the nucleus without the requirement of further truncation. Calpain family members, calpain 1, calpain 5 and calpain 10 show altered expression in Htt-transfected cells. Interestingly, calpain 10 localizes to the cytoplasm and the nucleus while the activated form is found only in the nucleus. Analysis of striatum and cortex from a HD mouse model supports the involvement of specific calpain family members in HD pathogenesis. These results suggest that aberrant activation of calpains alter Htt proteolysis and may play a role in the pathogenesis of HD.

Supported by: NIH RO1 NS40251A (LME), Huntington’s Disease Society of America, Hereditary Disease Foundation (LME and MRH) and Muscular Dystrophy Association (L.M.E.) and NIH F32 NS43937-01 (J.G.), and the Canadian Institute of Health Research (CIHR).

ABSTRACT 078-LATE-BREAKING RESEARCH: Behavioral manifestations of high extracellular dopamine in HdhQ92 knock-in mice.

Michel Cyr, Marc G. Caron (HHMI, Dept. of Cell Biology, Center for Models of Human Disease, IGSP, Duke University, Durham, NC, 27710, USA)

An expansion in the CAG repeat of the huntingtin gene underlie the development of Huntington’s disease (HD). Whether other molecular events might contribute to the susceptibility and/or onset of HD pathogenesis has remained unknown. Clinical studies reveal that the majority of HD individuals exhibit neuropsychiatric symptoms which can be corrected to some extend by antipsychotic drugs blocking dopamine receptors, and that these symptoms usually precede the onset of overt manifestations of HD. To examine the potential contributing role of the dopamine system in the emergence of pathological conditions in HD, we generate a double mutant mouse strain with both striatal hyperdopaminergic tone and endogenous expression of the human huntingtin gene containing 92 CAG repeats (HdhQ92). This strain has been produce by mating a dopamine transporter knockout (DAT KO) mouse, which exhibit elevated extracellular dopamine levels in the striatum and locomotor hyperactivity in a novel environment, to an HdhQ92 knock-in mouse that does not exhibit signs of neurodegeneration or behavioral abnormalities. Similar mortality rates were observed in DAT KO/HdhQ92 knock-in and DAT KO/Hdh WT mice during a period of one year. When activity was evaluated using an automated Omnitech Digiscan apparatus, the locomotion (horizontal activity) of both mice was enhanced to levels comparable to the one previously reported in DAT KO mice alone. However, at 4-months of age, this locomotor hyperactivity was progressively reduced in DAT KO/HdhQ92 knock-in mutants while in contrast, rearing (vertical activity) and stereotypy time were increased in comparison to DAT KO/HdhQ92 knock-in mice. These data suggest that striatal neuronal overactivation in combination with a mutant huntingtin protein can trigger alterations in the control of locomotor behavior.

ABSTRACT 083-LATE-BREAKING RESEARCH: Cell culture screen identifies caspase inhibitors as potential drugs for the treatment of Huntington’s disease

Authors: C.T.Aiken1; A.J.Tobin1,2,3; E.S.Schweitzer1,21Brain Research Institute, 2Depts. of Physiol. Sci., and 3Neurology, UCLA, Los Angeles, CA, USA

In search of potential treatments for Huntington’s disease (HD), we developed a cell-based, medium-throughput drug screen assay. Using this assay, we performed a blind screen of 1,040 compounds, compiled by the NINDS, to determine their abilities to protect expanded repeat huntingtin (HttQ103)-expressing PC12 cells from death. We quantified cell death, in 96-well plates, using a colorimetric assay that measures the release of the soluble cytoplasmic enzyme lactate dehydrogenase (LDH) into the culture medium. 18 compounds (2% of the 1,040 drugs tested) completely protected the HttQ103-expressing cells in our assay. An additional 5% were partially protective. 12% of the compounds tested were toxic to both HttQ103-expressing and non-expressing cells, and the majority of the compounds (81%) had no measurable effect. Several of our hits overlapped with those obtained by other investigators using different HD model systems, including those of whole organisms: C. el! egans and Drosophila.Another general caspase inhibitor, Z-VAD-FMK, was identified among the completely protective compounds from the library. It remains uncertain whether neurons undergo apoptosis or necrosis in HD, but the protective effects of caspase inhibitors in our cell system support apoptosis as the mechanism of cell death caused by HttQ103. We have therefore examined the effects of more specific caspase inhibitors to identify particular caspases involved and their relative importance. We have found that none of the narrow-spectrum inhibitors protects as completely as BOC-D-FMK or Z-VAD-FMK. An understanding of how these caspases contribute to cell death caused by the expression of HttQ103 will provide insight into the pathogenic mechanisms of Huntington’s disease and permit a more rational search for effective treatments.

Funded by NINDS and HDF.

TOPIC: ANIMAL MODEL EXPERIMENTAL THERAPEUTICS

ABSTRACT 003: A strategy for drug discovery in Huntington’s disease

Jamot Laure, Pruss Rebecca.(TROPHOS, Parc Scientifique de Luminy, Luminy Biotech Entreprises, Case 931, 13288 Marseille Cedex 9, France).

TROPHOS is a drug discovery biotech company. We are using high-throughput screening of a diverse, drug-like chemical collection on primary neurons to search for drug candidates for neurodegenerative diseases. TROPHOS has programs on motor neuron diseases (both SMA and ALS), Alzheimer’s disease and Huntington’s disease (HD). Since the mechanisms of degeneration and death are not fully understood in these diseases, it is not always possible to use specific molecules as screening targets. TROPHOS aims to target the pathways involved in the degeneration of neurons; molecules may therefore be identified that block events at many different stages in the degenerative process. All of the neurodegenerative diseases affect specific populations of neurons. The reasons for this are not clear, particularly since the mutant proteins involved (e.g. SMN, SOD1, Htt) are expressed ubiquitously. We assume that there are pathways or targets that are unique to these populations, making them susceptible to the disease. TROPHOS has therefore taken the approach of carrying out its screening on the population of neurons affected by the disease (striatal neurons for HD). In order to carry out screening, we have developed a new cytometer. This fluorescence-based analyser takes snapshot images of each individual culture well and can measure changes at the single cell level. The analyser is part of an integrated robotics system that has been optimised for dealing with primary neuronal cultures. For HD, TROPHOS has developed an original model based on cultured striatal neurons transfected with vectors expressing mutant huntingtin (Htt). Expression of mutant Htt induces the death of 50% of cultured primary striatal neurons, whereas normal Htt has no effect on neuronal survival. Moreover, this cell death can be blocked by trophic factors. Hit profiling studies performed by TROPHOS using other models of neurodegeneration and in collaboration with academic laboratories will explore the mechanism of action of active molecules. To demonstrate activity in vivo, TROPHOS plans to use models that recapitulate the pathology associated with mutant huntingtin. We expect by a combination of these approaches to eventually bring to the clinic molecules that act at all levels of the disease pathway.

ABSTRACT 026: A dual role of adenosine A2A receptors in 3-nitropropionic acid-induced striatal lesions: implications for the neuroprotective potential of A2A antagonists

David BLUM, Marie-Christine GALAS, Annita PINTOR, Emmanuel BROUILLET, Catherine LEDENT, Christa E. MULLER, Kadiombo BANTUBUNGI, Mariangela GALLUZZO, David GALL, Laetitia CUVELIER, Anne-Sophie ROLLAND, Patrizia POPOLI, Serge N. SCHIFFMANN

Reduction of A2A receptor expression is one of the earliest events occurring both in Huntington’s disease patients and in mice overexpressing the N-terminal part of mutated huntingtin. Interestingly, increased activity of A2A receptors has been found in striatal cells prone to degenerate in experimental models of this neurodegenerative disease. However, the role of A2A receptors in the pathogenesis of Huntington’s Disease (HD) remains obscure. In the present study, using A2A-/- mice and pharmacological compounds in rat, we demonstrate that striatal neurodegeneration induced by the mitochondrial toxin 3-nitropropionic acid (3NP) is regulated by A2A receptors. Our results show that the striatal outcome induced by 3NP depends on a balance between the deleterious activity of presynaptic A2A receptors and the protective activity of postsynaptic A2A receptors. Moreover, microdyalysis data demonstrate that this balance is anatomically determined since the A2A presynaptic control on striatal glutamate release is absent within the posterior striatum. Therefore, since blockade of A2A receptors have differential effects on striatal cell death in vivo depending on its ability to modulate pre- over postsynaptic receptor activity, therapeutic use of A2A antagonists in Huntington’s as well as in other neurodegenerative diseases could exhibit a undesirable biphasic neuroprotective-“neurotoxic” effects.

ABSTRACT 038: PEI mediated In vivo gene transfer in transgenic Huntingtons Disease mice. Evaluation of transfection method, effect and biomarkersBjarke Naver, Anne Nørremølle, Lis Hasholt, Sven Asger Sørensen (Sections of Neurogenetics, Institute of Medical Biochemistry and Genetics, University of Copenhagen, Denmark), Morten Møller (Institute of Medical Anatomy – Dept. B, The Panum Institute, University of Copenhagen, Denmark), Charlotte Stub, Axel Kornerup (Center of Bioethics and Risk Assesment, Division of Laboratory Animal Science and Welfare, Department of Pharmacology and Pathobiology, The Royal Veterinary and Agricultural University, Denmark)

A potential treatment for Huntington’s Disease and other neurodegenerative diseases is gene therapy. The major aim of gene therapy is to deliver genetic material into cells to alter their function. The genetic sequence to be delivered can be a gene encoding a protein, which is missing or present in insufficient amounts. However sequences designed to counteract a disease-related effect are being increasingly investigated. Reducing huntingtin expression by use of the antisense principles is a straight forward strategy for treatment of HD.

We are investigating the possibility of using antisense RNA expression for inhibiting the expression of the HD gene. Inhibition of the translation of the HD gene mRNA has been shown in an in vitro translation system using an approximately 200 bp antisense sequence of the HD gene. Using this antisense sequence of the HD gene cloned into a plasmid expression vector we performed in vivo intra-cerebral transfections in R6/1 mice using the polyethyleneimine transfection system. We have studied the short term and long term effects of an HD antisense RNA expression plasmid vectors and HD sense RNA expression plasmid vectors. Inclusion formation as well as motor abilities in the R6/1 mice were studied. Both transfectins with the antisense and sense constructs resulted in the inhibition of inclusion formation. However long term antisense treated mice foot-printing did perform better in the foot-printing analysis, suggesting that sequence specific effects succeed the stress induced effects caused by the in vivo transfections.

ABSTRACT 043: Histone Deacetylase Inhibitors Prevent Oxidative Neuronal Death Independent of Expanded Polyglutamine Repeats via a Sp1-Dependent PathwayH. Ryu, J. H. Lee, A. Mwidau, and R. R. Ratan (Department of Neurology and the Program in Neuroscience, Harvard Medical School and Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115), A. Dedeoglu, R. J. Ferrante (Geriatric Research and Education and Clinical Center, Bedford VA Medical Center and Neurology, Pathology, and Psychiatry Departments, Boston University School of Medicine, Boston, MA 02118), B. A. Olofsson, J. Azizkhan-Clifford (Drexel University College of Medicine, Philadelphia, PA 19102), M. Escudero, E. Flemington (Department of Pathology, Tulane University School of Medicine, New Orleans, LA 70112)

Oxidative stress is believed to be an important mediator of neurodegeneration. However, the transcriptional pathways induced in neurons by oxidative stress that activate protective gene responses have yet to be fully delineated. Immunoprecipitation with an Sp1 antibody, followed by Western blot analysis with an acetyl-lysine antibody, revealed a statistically significant increase in Sp1 acetylation in response to HCA-induced oxidative stress in cortical neurons. We confirmed that the transcription factor Sp1 is acetylated by p300 in vitro. Histone deacetylase (HDAC) inhibitor, trichostatin A (TSA), augmented Sp1 acetylation, Sp1 DNA binding and Sp1-dependent gene expression and conferred resistance to oxidative stress-induced death in vitro. The structurally distinct HDAC inhibitors, sodium butyrate and suberoanilide hydroxamic acid (SAHA), also abrogated oxidative death in primary cortical neurons. We found that protection by sub-maximal concentrations of TSA is significantly reduced by the Sp1 antisense oligonucleotide (ODN) (p <.05), but not the mismatch ODN. This data implied that Sp1 activation is necessary for the protective effects of HDAC inhibitors to occur. Animals concomitantly treated with sodium butyrate and 3-nitropropionic acid (3-NP) had increased levels of acetylated Sp1, as compared to 3-NP treated animals alone. The increased in vivo acetylation of Sp1 induced by sodium butyrate treatment was associated with nearly complete protection from striatal 3-NP toxicity, as determined by Nissl staining and TUNEL labeling. The histopathological evaluation of 3-NP-induced striatal lesion-volumes showed significantly less tissue damage in sodium butyrate-treated mice than in PBS-treated mice. Together, these results demonstrate that HDAC inhibitors inhibit oxidative death independent of polyglutamine expansions by activating a Sp1-dependent adaptive response.

ABSTRACT 053: Use of Adult Bone Marrow Stem Cell Transplants to Counteract Cognitive Deficits in a Rat Model of Huntington’s Disease.

Gary L. Dunbar (BRAIN Center, Department of Psychology, Central Michigan University, Mt. Pleasant, MI 48859)and Laurent Lescaudron (Institut de Transplantation et de Rescherche en Transplantation, Universite de Nantes, 44035 Nantes, France)

The purpose of this study was to investigate possible therapeutic effects of autologous bone marrow transplantation in a rat model of Huntington’s Disease. Thirteen days after bilateral injections of quinolinic acid (QA) into the rat striatum, whole bone marrow was aspirated from the left tibia and bilaterally implanted into the damaged striatum of each rat. Autologous transplantation was used in order to mitigate possible immune responses to the transplanted cells. Thirteen days after transplantation, the ability of bone marrow derived stem cell transplants to reverse QA-induced learning deficits in the radial-arm water maze (RAWM) was examined. Visuospatial abilities and swim speed were also assessed. An additional group of rats was implanted with labeled cells in order to study cell migration, integration, and differentiation within the damaged striatum. Our results indicated that QA caused a significant increase in both working- and reference-memory errors in the RAWM task, and that the transplants significantly reduced the number of working memory errors. Transplanted cells migrated throughout the whole striatum, the fronto-parietal cortex and the nucleus accumbens. Some of the labeled cells were visible in the capillaries, suggesting a migration through the brain blood barrier. Most of the transplanted cells did not express any neural phenotypes and appeared quite primitive. However, a very small percentage of the transplanted cells were positive for nestin, beta-tubulin-III, and GAD, but none were positive for GFAP. Thirty seven days post-transplantation, evidence of metabolic activity within the transplanted cells was evident. Because the therapeutic effects of the transplanted stem cells were observed as early as 10 days post-transplantation and because very few of transplanted cells differentiated into neurons, we hypothesize that the release of growth factors by the transplanted cells allowed the surviving cells in the host tissue to function more efficiently or promote other compensatory responses in the damaged brain.

ABSTRACT 086-LATE-BREAKING RESEARCH: Cop-1 vaccination reduces motor function deficits and increases life expectancy in a transgenic mouse model of Huntington’s disease

Hadas Schori, Michal Schwartz

Immune system was recently shown to play a crucial role in protecting against loss of neurons after acute or chronic insults to the central nervous system (CNS). Moreover, appropriately-controlled boosting of the T cell-mediated autoimmune response causes the cells to recognize and home to their specific antigens at the lesion site, where they regulate microglia behaviour and thereby protect neurons from further degeneration. Cop-1, a synthetic antigen of wide specificity, can elicit this neuroprotective response without inducing an autoimmune disease. Thus, vaccination with Cop-1 promotes the survival of neurons exposed to a toxic excess of the neurotransmitter glutamate, a major risk factor in neurodegenerative diseases. The neuroprotective effect is dependent on activation and proliferation of Cop-1-specific T-cells, which secrete the cytokine IFN-g. Repeated injections of a single dose of Cop-1 at 4-week intervals maintained the neuroprotective effect in a model of glutamate toxicity in the mouse eye. We tested this dose and regimen using Cop-1 in R6/2 mice, a transgenic model for Huntington’s disease (HD), an autosomal dominant neurodegenerative disorder whose symptoms are caused by the loss of neurons in the basal ganglia of the brain. Cop-1 was injected into 45-day-old R6/2 mice and repeated every 4 weeks. Motor functions (tested on a Rotarod), weight loss, time of disease onset, and life expectancy were evaluated. Vaccination with Cop-1 resulted in significant improvement of the motor performance and delayed the onset of disease and mortality. These results demonstrate that the neuroprotective effect achieved by boosting of the immune system with a suitable antigen in an appropriate regimen attenuates neurological decline in HD transgenic mice.

TOPIC: CLINICAL CHARACTERISTICS

ABSTRACT 016: Subcortical Mild Cognitive Impairment: a novel concept of mental dysfunctionBonelli RM, Hödl A, Otti DV, Lahousen T, Wenning GK, Kapfhammer HP

Background: The term “subcortical dementia” refers to the behavioural symptom of degenerative disorders involving primarily subcortical structures. It is characterized by slowness of mental processing, forgetfulness, impaired cognition, apathy, and depression. Mild cognitive impairment (MCI) is a stage between “normal” aging and dementia. Common activities of daily living are intact, but there may be subtle impairment in complex, instrumental activities of daily living. Cortical MCI (cMCI) is said to be prodromal for Alzheimer’s disease. Subcortical MCI (sMCI), on the other hand, has never been proposed so far. Patients & Methods: Research definitions of MCI using the global deterioration scale (GDS) stage 3 were applied to diagnose MCI in 61 patients with a MMSE > 23. Out of these, 41 patients suffered from degenerative disorders classically attributed to subcortical dementia (i.e. Huntington’s disease, Parkinson’s disease, multiple system atrophy, progressive supranuclear palsy, multiple sclerosis). These patients (sMCI) were compared to 20 patients with cMCI (diagnosed by SPECT, MRI, and CSF parameters) and 30 age and sex matched controls (GDS stage 1). All subjects underwent an intensive neuropsychological test battery. Results: We found a significant difference between the sMCI group and controls in calculating, figural but not verbal memory, trail making test B, perseverations, executive functions, and verbal fluency. In contrast, cortical MCI showed more deficits in verbal memory and design fluency, but not in perseverations. When the two MCI groups were compared it turned out that perseverations (more impaired in sMCI) were a significant factor as well as orientation, design fluency, and complex figure organization (more impaired in cMCI).

Discussion: Our data suggest that sMCI is common in neurodegenerative disorders affecting subcortical circuitry regardless of underlying etiology. In addition, sMCI and cMCI can be discriminated in terms of their neuropsychological deficit profile. Further studies are required to determine the evolution of patients with sMCI versus cMCI.

ABSTRACT 018: Nature and development of HD in a nursing home population: The further development of the BOSH rating scaleHans Claus (Stichting Duyn & Rhyn, location Overduin, Katwijk, The Netherlands), Reinier Timman (Erasmus Medical Center Rotterdam - Medical Psychology & Psychotherapy, The Netherlands; Leiden University Medical Centre - Neurology, Leiden, The Netherlands), Henk Slingerland (Atlant Zorggroep, location Hullenoord, Beekbergen, The Netherlands), Marijanne van der Schalk (R.K. Zorgcentrum Sint Jacob, Amsterdam, The Netherlands), Saskia Demeulenaere (Home Marjorie, Heist-op-den-berg, Belgium), Aad Tibben (Leiden University Medical Centre - Neurology, Leiden, The Netherlands)

The BOSH (Behavior Observation Scale Huntington) was developed for a more detailed description of the HD-patient in final phases of the disease. The scale was validated in four nursing homes with a specialized ward for HD-patients. For 91 patients the BOSH was administered twice independently by different observers. Test- retest reliabilities were satisfactory. The BOSH contains 32 items. From principal component analysis three subscales emerged with adequate internal reliability: 1) ADL (Activities of Daily Living), 2) social cognitive functioning and 3) aggression and rigidity. The hospitalization period seems not a good indicator for disease duration. We hypothesize that ADL is a better indicator. A non-linear relation was found between aggression and rigidity and ADL. This relation can be interpreted as aggression and rigidity becoming more frequent as the disease progresses in the first stages, until deterioration causes these behaviors to diminish. A second non-linear relation was found between social-cognitive capabilities and ADL, which suggests that these capabilities deteriorate more rapidly in later stages of the disease. In a follow-up study the relation between ADL and disease duration will be ascertained more precisely. Also, the relation with scores on the UHDRS (Unified Huntington’s Disease Rating Scale), the GMS (Geriatric Mental State Schedule), the pre-morbid personality (NEO-FFI) and the social support (SSL) will be explored. More insight into the dynamics of the disease and its progression enables the nursing homes staff to better adjust to the individual needs of HD-patients.

ABSTRACT 024:Objective assessment of akinesia and bradykinesia in Huntington’s diseaseJeroen P.P. van Vugt, Karlijne K.E. Piet, Liesbeth J. Vink, Sabine Siesling, Aeilko H. Zwinderman*, Huub A.M. Middelkoop, Raymund A.C. Roos (Department of Neurology and *Department of Medical Statistics, Leiden University Medical Center, Leiden, The Netherlands)Introduction - Functional disability of patients with Huntington’s disease (HD) is determined by impairment of voluntary motor function rather than the presence of chorea. Only few attempts have been made to quantify these parkinsonian symptoms. Aims - To investigate (1) which clinical factors (e.g. clinical parkinsonism, cognitive impairment, use of medication) determine quantified motor slowness in HD, and (2) whether quantified motor slowness is a responsive measure of functionally important changes during prospective evaluation of HD patients.Methods - Using a simple reaction time paradigm, we measured the time needed for movement initiation (akinesia) and execution (bradykinesia) in 76 HD patients and 127 controls. Furthermore, patients were rated clinically using UHDRS subscores for eye movements, impairment of voluntary movements (i.e. parkinsonism), dyskinesias, posture and gait, and depression, the Mini-Mental State Examination, and Shoulson and Fahn’s Total Functional Capacity. Results - Quantified akinesia and bradykinesia were already evident in early stages and increased linearly with increasing disease stage. Quantified motor slowness correlated with clinical parkinsonism, but also with cognitive impairment and medication use (i.e. neuroleptics and sedatives). In patients without severe cognitive impairment, quantified motor slowness reflected clinical parkinsonism more purely. During 1.9 years follow-up (range 0.8-3.8 years), quantified akinesia and bradykinesia progressed concomitantly with progression of clinically rated parkinsonism, cognitive impairment and functional disability. However, rate of change in motor slowness did not discriminate between patients whose disease stage remained stable and those whose disease stage progressed. Conclusion - We conclude that the reaction time paradigm may be used to quantify akinesia and bradykinesia in HD, at least in patients without severe cognitive impairment. In patients, reaction and movement times became worse over time. However during our relatively short follow-up period, this progression of quantified motor slowness was not marked enough to detect functionally important changes. Financial disclosure: J.P.P. van Vugt received sponsoring for his Ph.D. thesis from Janssen-Cilag B.V., GlaxoSmithKline, and Servier Nederland B.V. ABSTRACT 028: Somatoform disorders as the initial presentation of Huntington diseaseVicki L. Wheelock, Ron Risley, Teresa Tempkin (University of California Davis Medical Center, 4860 Y Street, Sacramento, CA 95817), Kathleen Baynes (Center for Neuroscience, University of California Davis, 1544 Newton Court, Davis, CA 95616)

Objective: To report three patients manifesting somatoform disorders as the initial presentation of HD.Background: The most widely accepted diagnosis of manifest HD is the unequivocal presence of an extrapyramidal movement disorder. Families and experienced clinicians recognize that psychiatric or cognitive symptoms may precede the onset of motor symptoms. Somatoform disorders are characterized by a focus on physical symptoms in individuals who claim symptoms or impairment not fully supported by physical examination or by laboratory or imaging studies. There have been no previous reports of somatoform disorders associated with HD. Method: Case report Results: Three women presented to the HD clinic seeking treatment for neurological symptoms that were atypical for HD, including pseudoseizures (2), blurred vision (2), dizziness, bizarre gait disorder (2), sensory loss, and unilateral weakness. Neurological examination in each revealed nonphysiological findings. UHDRS total motor score was zero. EEG confirmed non-epileptic events and MRI imaging of the brain (3) and spine (2) was normal. Genetic testing revealed expanded CAG alleles in each patient. Neuropsychological testing demonstrated subtle frontal and executive dysfunction. MMPI revealed strong somatic preoccupation in two patients and mild depression in one. These patients reported physical symptoms starting before age 30 and experienced between 4-6 neurological symptoms, 2-4 pain symptoms, 2 sexual symptoms, and 1-3 gastrointestinal symptoms consistent with DSM-IV-TR criteria for somatization disorder in two and undifferentiated somatoform disorder in the third. Conclusions: Somatoform disorders can precede the onset of extrapyramidal motor abnormalities in individuals with the HD CAG expansion. Individuals at risk for HD commonly experience the distress of observing progressive, debilitating neurological changes in one or more family members. Degeneration of striatal associative pathways may result in somatoform disorders as a psychiatric manifestation of HD. Identification of

somatoform disorders has important implications both for predictive testing and for appropriate and effective medical management.

ABSTRACT 031: Does the clinical spectrum of Huntington’s disease include tremor?

Dr Neil Mahant, Ms Shanthi Graham, Dr Elizabeth A McCusker (Westmead Hospital, Sydney, Australia) Objectives: To identify the spectrum of clinical features and forms of tremor in Huntington’s disease (HD).

Methods: Our clinic provides care for patients at all stages of HD at no cost to patients.Consecutive patients were examined and rated according to standardised assessment scales, recorded on videotape where possible. The clinical features of five additional patients, not included in the consecutive clinic sample, are also described.

Results: There were 72 patients during the study period, 20 of whom did not consent to participate or were excluded due to the presence of a significant psychiatric or other co-morbidity.The average age of the 52 participants was 52±12 years, with the severity of HD ranging from almost no sign of disease to incapacitated. The clinical scores, including the dystonia and chorea scores, were normally distributed.Rhythmical movement disorders were present in 15 subjects (29%): postural tremor (4), kinetic tremor (3), postural and kinetic tremor (6), clonus due to severe spasticity (1) and stereotypy of the leg (1). Patients with tremor had a greater CAG repeat length (p=0.006), motor disorder (p=0.05) and level of disability (p=0.007) than patients without tremor. Myoclonus was present in 1 patient, and other atypical movement disorders were not seen. One of the five additional patients had an irregular head tremor and torticollis; the other patients had a postural/kinetic tremor of the hands (one also had rest tremor) in association with prominent akinesia.

Conclusions: The phenotypic expression of HD is complex, representing a spectrum from chorea-predominant to akinesia-predominant disease. The ‘Westphal’ variant represents one end of the spectrum rather than a separate entity. Tremor is more common than other atypical features, but may in some cases be drug induced.

ABSTRACT 033: Stereotypic Leg Movements in a Family Member of Huntington’s Disease

Din-E Shan, Bing-Wen Soong, Neurological Institute, Taipei Veterans General Hospital, and School of Medicine National Yang-Ming University, Taipei, Taiwan, ROC

We reported a 46-year-old woman from a family with genetically proved Huntington’s disease. Her father had involuntary movements and expired at the age of 57. One of her brothers and one sister had involuntary movements for 8 years and 7 years, respectively. Both had onset at about 48-year-old. The patient visited our outpatient clinic in 1996 due to involuntary movements of the right big toe for half a year. She also felt stiff of the right leg, with frequent leg cramp. She did not felt any urge to move the limbs, and she could not suppress the movements on volition. There were no precipitating factors. Hyperthyroidism was told at 20-year-old. Neurological examination revealed the presence of repetitive, involuntary, arrhythmic, and brief dorsiflexion of the right big toe and fanning of other toes. In 1998 increasing blinking of both eyes was noted. She began to feel exacerbated nocturnal leg pain and jerking of the legs while sleeping. The leg movements became more periodic, occurring every 17 to 30 seconds. Most laboratory tests showed normal results. Thyroid hormones were slightly elevated. PCR analysis of the IT15 gene demonstrated the presence of expanded CAG repeat with an upper allele of 40 repeats in the sister, but not in the patient. We concluded that the patient did not carry Huntington disease gene mutation. The movements that she had appeared to be a form of periodic leg movements and had a benign course. However, some questions remained to be answered. Was the periodic movements a coincident finding or related to other gene trait in the family? Did the movements indicate dopaminergic deficiency, as in the cases of Parkinson disease with periodic leg movments? Why did the movements appear in the day and not limit in sleep? More investigations are required to answer these questions.(no personal financial relationship)

ABSTRACT 034: Abstract_title: Psychomotor slowing and memory disturbances over a three-year period in presymptomatic’ carriers for Huntington’s Disease

Marie-Noëlle W. Witjes-Ané, Bart Mertens, Jeroen P.P. van Vugt, Gert-Jan B. van Ommen, Raymund A.C. Roos (Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, the Netherland) Jan B.K. Lanser (Rijnland Hospital, Delftzichtweg 2, 2402 NB Alphen a/d Rijn, The Netherlands)

Background: Predictive testing in Huntington’s Disease (HD) nowadays allows long-term follow-up of ‘presymptomatic’ carriers to eventually determine the sequence of appearance in symptoms. Research into the first manifestations of Huntington’s Disease (HD) is essential to improve the diagnostic process. As the expectation of new effective (farmaco)therapeutic strategies will be developed, it becomes even more important to assess onset of clinical disease and to review the progression of the disease. Because of the disparate findings reported in the literature, we focussed on cognitive and motor functioning.Methods: Thirty-three carriers and 73 non-carriers, who had undergone predictive testing, attended three occasions with 18 months interval. Single-blind assessment of the Unified Huntington’s Disease Rating Scale (UHDRS) and an extensive neuropsychological battery was performed.Results: Carriers performed consistently worse than non-carriers over time on memory tasks (auditory verbal learning task and visual retention task), timed psychomotor tasks and quantified motor assessment. The decline seen in timed psychomotor tasks involving a sequencing aspect was predominantly due to deficits in mental speed rather than motor or cognitive disturbances. Furthermore, we found that estimated onset age (based on CAG-repeat length) had significant influence on changes in the Mini Mental State Examination.Conclusions: Quantified motor assessment, psychomotor tasks with a sequencing component, auditory verbal learning tasks and visual retention tasks might be vulnerable in the preclinical phase of HD. Carriers might have reached a ‘zone of onset’ as psychomotor and memory decline could be seen as precursors of manifest HD. The findings suggest that different aspects of cognitive changes in HD carriers do not evolve uniformly. Furthermore, we conclude that psychomotor tasks and quantified motor assessment are a useful addition to neurological examination in early HD.

ABSTRACT 035: Assessment of change in cognitive function in Huntington’s Disease

Julie S Snowden, Jennifer C Thompson (Greater Manchester Neuroscience Centre, Hope Hospital, Salford, UK), David Craufurd (University of Manchester, Department of Medical Genetics, Manchester, UK).

Cognitive impairment is a core feature of HD. Evaluation of efficacy of novel therapies for HD depends upon the availability of reliable neuropsychological assessment instruments, sensitive to changes in cognitive function. Neuropsychological studies of HD have highlighted the principle domains of cognitive impairment: psychomotor slowing, executive deficits and memory impairment. Neuropsychological tests that are sensitive to the presence of HD are well established. However, longitudinal studies of the natural evolution of cognitive change have been relatively limited, so that it is much less clear which tests are most sensitive to change in cognition over time. This study reviews existing longitudinal studies from the US and Europe, examines the types of measures that were used and identifies those measures demonstrated to be most sensitive to change. An essentially consistent pattern of findings is revealed across studies. Psychomotor tasks, such as the Stroop test and Trail making, are most sensitive to change. Non-timed executive tests, such as the Wisconsin Card Sorting test, show poor sensitivity to change. Measures of memory and general intellectual function are also relatively poor markers of change. Our own study of asymptomatic people who carry the HD mutation reveals complementary findings: cognitively undemanding psychomotor tasks are most sensitive to the earliest changes of HD. Differential sensitivity of tests in part reflects intrinsic properties of the tests, such as vulnerability to practice effects and the gradation of the scale along which performance is measured. However, we argue that it also reflects genuine differences in sensitivity to the evolution of HD. We argue that psychomotor performance taps striatal function, and is a fundamental marker of severity of HD. We argue that simple rather than sophisticated, cognitively demanding neuropsychological tests provide the best tools for measuring change in HD and for use in clinical trials. No personal financial relationship to any product or sponsor

ABSTRACT 041: Does the cause of functional disability change with disease progression in patients with Huntington’s disease?

Yvette A.M. Grimbergen, Jeroen P.P. van Vugt, Douwe Pons, Raymond A.C. Roos (department of Neurology, Leiden University Medical Centre, Leiden, The Netherlands), Sheila A. Simpson (department of Medical Genetics, Medical School, Aberdeen Royal Infirmary, Aberdeen, Scotland)

Introduction: The aim of this study was to explore what clinical symptoms determine functional disability and functional decline in a European cohort of patients with Huntington’s disease (HD).Methods: Since the introduction of the Unified Huntington’s Disease Rating Scale (UHDRS) in 1994, Huntington’s disease patients in Leiden and Aberdeen were rated when they visited the outpatient clinic.These scores were gathered in a database.In the present analysis we included patients that were assessed two times or more, at least 6 months apart.Assessments missing significant parts of the motor or functional scores were excluded. Pearson’s correlation followed by stepwise multiple regression analysis was used to determine what symptoms correlated with (changes in) functional disability.Results: Baseline assessments were available from 283 patients.Of these, 136 were excluded due to missing follow-up. No medical history was available from thirty-three patients, while another 15 patients were excluded due to incomplete motor or functional scores. Therefore, ninety-nine patients were included in total (50 men; mean age 46.3 years (range 20-74); mean disease duration 7.3 years (range 0.2-23.2)). At baseline, functional disability correlated best with impairment of voluntary movements, posture and gait and with the UHDRS total motor score. Weaker correlations were found with verbal fluency, symbol digit and Stroop tasks.The mean follow-up time was 3.1±1.74 years (range 0.5-8.04). At follow-up, functional disability correlated best with impairment on cognitive tasks (i.e. symbol digit modalities test and MMSE). The rate of decline in total functional capacity (TFC) was 0.66±0.94 units/year.The rate of functional decline did not correlate significantly with rate of decline in any specific symptom.Conclusion: The present data suggest that functional impairment may primarily be determined by motor impairment in earlier stages of HD, but by cognitive impairment in later stages.

ABSTRACT 048: Changes in sexual behaviour associated with Huntington’s disease

David Craufurd (University of Manchester), Jennifer Thompson, Julie S Snowden (Greater Manchester Neuroscience Centre).

AIMS: Previous reports in the medical literature, including that of Huntington himself, have suggested that Huntington’s disease (HD) is associated with hypersexuality. However, the only study to date which examined the issue systematically using modern research methods found no evidence to support this (Fedoroff et al., 1994). We therefore investigated libido and sexual behaviour as part of a broader study of behaviour in HD patients.METHODS: 134 patients attending an HD management clinic were interviewed with their principle carer using the Problem Behaviours Assessment for Huntington’s disease.RESULTS: Loss of libido was reported by 61.8% of patients for whom reliable information (corroborated by the partner) was obtained. Loss of libido correlated significantly with the PBA-HD apathy score (rho 0.409, p<0.01) and inversely with Total Functional Capacity score (rho –0.456, p<0.0001). Sexually disinhibited behaviour was reported by carers in only 6/104 individuals (3 male, 3 female; 5.8%), while demanding or persistent sexual behaviour was described in just 4/81 cases (3 male, 1 female; 4.9%). Hypersexual behaviours were associated with a behavioural profile characterised by irritability, mental inflexibility and obsessive-compulsive or perseverative behaviours.CONCLUSIONS: Hypersexual behaviour is rare in HD and affects a small minority of cases only. Loss of libido and hyposexuality are much more common and correlate strongly with other measures of disease progression. The theoretical implications will be discussed.No personal financial relationship to product or sponsor.

ABSTRACT 050: Walk and Drink Time; Objective measures of motor progression in Huntington’s Disease.

Elizabeth M Howard (St Mary’s Hospital) Jennifer C Thompson,Julie S Snowdon (Greater Manchester Neuroscience Centre) David Craufurd (University of Manchester)

Reliable measures of motor progression in Huntington’s Disease are essential for the evaluation of new treatments now being tested. The Unified Huntington’s Disease Rating Scale (UHDRS) is generally accepted as a sensitive tool for this purpose, (Seisling et al.,1998) with good inter-rater reliability (Huntington’s Study Group., 1996). In practice, however the utility of the UHDRS for longitudinal studies may be limited, particularly where frequent assessments are required. Scores may be affected by variation in chorea from one appointment to the next due to the impact of emotional state on the severity of involuntary movements. The gradual decline of involuntary movements in late stage disease may also cloud the issue. In addition to the UHDRS motor scale we have used two further measures for evaluation of motor progression of HD. Both are simple quantitative assessments which require no special equipment to carry out. They are also easily reproducible by non-medical personnel. Patients are timed as they walk a fixed distance (including a 180 degree turn) as fast as they can. They are also timed as they drink 130mls of water as fast as possible. Data were collected from 97 individuals affected by HD of whom 47 were male and 50 female. Ages ranged from 22 to 81 years (mean 48.3,SD11.9). Duration of illness varied from 1 to 21 years (mean 8.1,SD 4.3). The shortest walk time was 10 seconds and the longest 85 seconds. There was even greater variability in drink time from 5 to 330 seconds. There was a highly significant correlation between duration of illness and both walk time (rho 0.562, p<0.001) and drink time (rho 0.279, p<0.01) as well as between the two measures themselves(rho 0.449, p<0.001). These new measures can therefore be used as simple and reliable additions to the UHDRS motor scale when considering longitudinal outcomes.

ABSTRACT 055: Social cognition in Huntington’s disease and frontotemporal dementia

Jennifer C Thompson, Julie S Snowden, Cheryl L Stopford, David Neary (Greater Manchester Neuroscience Centre, Hope Hospital, Salford, United Kingdom) David Craufurd (Department of Clinical Genetics, St Mary’s Hospital, Manchester, United Kingdom)

Altered social conduct and a breakdown in interpersonal relationships are prominent features of HD. Patients are often described as being self-centred, demanding, mentally inflexible and lacking in empathy for others. The factors underlying this breakdown in social cognition remain poorly understood. A previous study (Snowden et al, 2003) investigated the ability of HD and frontotemporal dementia patients (FTD) to interpret social situations and ascribe mental states to others. Although both patient groups were impaired on these tasks, the reasons underlying this differed. Typically, FTD patients failed to make inferences about social situations or the mental states of others. By contrast, HD patients tended to draw inappropriate inferences. Studying social cognition in individuals with general cognitive impairment can be problematic, because social cognition tests typically make demands on other cognitive abilities, such as memory and attention. Thus, failure may reflect impaired cognitive abilities, rather than a specific breakdown in social reasoning. The aim of this study was to further investigate the ability of HD and FTD patients to make social inferences. A novel social cognition task was developed, which aimed to minimise demands on other cognitive functions. Patients were presented with simple, illustrated vignettes that depicted everyday social situations and were required to answer a series of questions tapping their ability to understand the beliefs, thoughts and feelings of the characters depicted, and to draw inferences about the social situations, which necessitated going beyond the information given. The results demonstrated that although both patient groups were impaired relative to controls, the pattern of errors differed between the two groups. As in our previous study, the HD patients were particularly impaired in the ability to draw appropriate inferences from social situations. The finding that HD patients make faulty social inferences has implications for understanding the breakdown in social behaviour in HD.

ABSTRACT 069: Defining Progression in Huntington DiseaseDonald S. Higgins (Albany Medical College, 47 New Scotland Ave/MC136, Albany, NY 12208), Sudeshna Adak, Samit Paul (GE India Technology Centre Pvt Ltd, Export Promotion Industrial Park - Phase 2, Hoodi Village, Whitefield Road, Bangalore - 560 066 - India), Kati Illouz, Will Gorman (General Electric Global Research, One Research Circle, Niskayuna, NY 12309)

The TFC is a 13-point scale assessing a range of abilities in individuals with Huntington disease. Change in the TFC has been the primary outcome measure in many HD trials. With a modest dynamic range and substantial “floor/ceiling” effects, the TFC may be insensitive to modest yet relevant change. Incorporation of additional measures from the UHDRS may enhance this sensitivity. A decision tree analysis was applied to a series of 133 subjects with at least 2 visits (2-12 visits) and complete UHDRS records (58 M/75 F) followed for 1.0 ± 0.5 yrs (0.2 – 2.9 yrs). Rules were generated using absolute and change in 3 composite scores (motor, cognitive and functional), TFC and Independence scale (IS) that best correlated with physician assessment of progression (worse on question 78). The rules suggest that TFC and IS figure prominently in defining disease stage, while changes in motor, cognitive and functional performance are more useful in defining progression within each disease stage. Classification accuracy was greatest when absolute and difference measures were included (88%). Less dramatic collapse of the UHDRS (19 subscales) revealed a considerable role for chorea and balance in determining progression with a smaller contribution from difference measures, TFC and IS. Additional dimensions improved classification accuracy with the best performance incorporating absolute and difference measures (> 91%). Validation of clinically meaningful versions of these rules and assessment of rule performance in defining progression within various stages of HD is ongoing with planned application to a larger sample. This preliminary investigation suggests that progression in HD is more effectively assessed considering multiple components of the UHDRS, not simply the TFC. A composite measure, incorporating these dimensions would likely increase the precision with which progression is defined and treatment efficacy determined, thereby hastening the discovery of disease modifying therapies. No conflicts to disclose.

ABSTRACT 070: Safety and Feasibility of the Prospective Huntington At Risk Observational Study (PHAROS): A Progress ReportHuntington Study Group/PHAROS Investigators (presenter: Elise P. Kayson, University of Rochester; Rochester, NY)

Background & Objective: The Prospective Huntington At Risk Observational Study (PHAROS) was designed to objectively determine the earliest and most specific clinical signs of Huntington’s disease (HD) in a population of unaffected adults at risk for HD who chose not to undergo predictive DNA testing. A major goal of PHAROS is to assess the safety and feasibility for future clinical trials aimed at forestalling the onset of HD.Design/Methods: Eligible PHAROS research participants include unaffected adults (26 to 55 years old) at immediate risk for HD (a parent or sibling affected by HD) who agree to provide a blood sample for a confidential analysis of their HD gene status (the individual results of which will never be disclosed) and be clinically evaluated every 9 months by investigators who also remain unaware of gene status. Research participants and investigators are asked to report relevant events that may have a bearing on the safety and feasibility of PHAROS. Accrued reportable events are reviewed and evaluated periodically by an Event Monitoring Committee.Results: Since July 1999, 863 consenting adults (age 41.5 ± 7.5 years; mean ± sd) have been enrolled at 43 PHAROS research sites in the United States and Canada and have been followed for 2.2 ± 1.0 years. As of April 1, 2003, 141 reportable events have involved 115 (13.3%) participants, including 40 (2.1% annual incidence) who developed new-onset depression; 36 (1.9%) who were hospitalized for any medical reason; 28 (1.5%) who withdrew research participation; 23 (1.2%) who required a new mental health evaluation; and 2 (0.1%) who died (not suicide). No participant reported a breach of confidentiality or disclosure of HD gene status.Conclusions: The overall number and types of reportable events are in keeping with our expectations for this long-term study, including the annual incidence of events. So far, Withdrawal from PHAROS has been less than our projected 5% annual rate. The 2.1% annual incidence of new-onset depression approximates the 1.6% annual incidence reported in a general medical population (J Fam Practice 2000; 49:68-72). To date, PHAROS is a safe and feasible research undertaking. Financial Disclosure: The authors do not have any personal financial interest in this research.

ABSTRACT 072: Clinical predictors of neuropathological severity: a prospective autopsy study of 100 individuals with Huntington's disease

Adam Rosenblatt, M.D., Margaret H. Abbott, M.P.H., R.N., Lisa M.Gourley, M.A., Juan C. Troncoso, M.D., Russell L. Margolis, M.D., Jason Brandt, Ph.D., Christopher A Ross, M.D., Ph.D. (Johns Hopkins University School of Medicine, 600 North Wolfe Street, Baltimore, MD 21287)

Subjects were followed in the Longitudinal Core Study of the Baltimore Huntington’s Disease Center and given annual neurologic, cognitive, and psychiatric exams. Postmortem Neuropathological grade was assigned using the system of Vonsattel et al. We examined the correlations between the neuropathological grade and scores on the Quantified Neurological Exam (QNE) and its chorea and motor impairment subscales, the Mini-Mental State Exam (MMSE), the HD Activites of Daily Living (ADL) Scale, and a number of demographic variables (CAG number, age of onset, age at death, disease duration) in 100 subjects who had been examined within 1000 days of death. All measures showed significant correlation with Vonsattel score except for chorea. The strongest effect was that of motor impairment score (r-squared=.351, p<.0001). In a step-wise correlation of clinical variables, motor impairment remained significant. The largest effect for a demographic variable was for age of onset (r-squared=.226, p<.0001). A partial correlation was significant between CAG number and Vonsattel grade, controlling for age at death or disease duration. Motor impairment appears to be a good clinical measure of neuronal cell loss, at least late in the course of HD, and therefore may prove useful in observational and treatment studies.

ABSTRACT 073: Risk factors for Osteoporosis in Women with Huntington’s Disease in a Long-term care Setting

Joseph Klager, Bruce Berman, Sybil Montas, Ayana Duckett, Jean Perri, Robert Wilkins, Anthony J. Lechich, MD (Terence Cardinal Cooke Health Care Center, Medical Administration, 1249 Fifth Avenue, New York, NY 10029)

BACKGROUND Female residents in a long-term care unit for Huntington’s Disease (HD) were observed to have a high prevalence of amenorrhea and oligomenorrhea. The risk factors for estrogen deficiency and loss of bone mass have not been studied in this population. METHODS Risk factors for osteoporosis were retrospectively reviewed in 24 women with HD whose ages ranged from 35 to 73 years. RESULTS Of the 24 women, one showed severe bone loss on chest X-Ray, 22% showed moderate bone loss, 22% showed mild bone loss and 48% showed no bone loss. 79% of the women were either amenorrheic or oligomenorrheic and 16% of the women were underweight according to BMI calculations. Dietary calcium intake on average exceeded recommended daily value by 1.49 times while dietary vitamin D intake exceeded the recommended daily value by an average of 1.25 times. 76% of residents were non-ambulatory. History of cigarette smoking was present in16% of the women surveyed. CONCLUSIONS 52% of the women surveyed showed some degree of bone loss on chest X-ray. This result suggests that women with HD residing in long-term care facilities may be at an increased risk for developing osteopenia or osteoporosis. Impaired ambulation ability and abnormal menstruation patterns were two prevalent risk factors observed in the women surveyed. Thus, pre- and post-menopausal estrogen deficiency along with a lack of weight-bearing mobility could be causing bone loss in some women with HD.To further investigate bone loss and its potential mechanisms in women with HD, a bone mineral density analysis via dual-energy x-ray absorptiometry (DXA) and a complete laboratory analysis of estrogen and markers of bone metabolism are necessary. Due to the movement disorder and the accompanying risk of fall and injury, assessing and maintaining bone health should be viewed as important for women with HD.

ABSTRACT 079: Peg insertion reflects dysfunction in Huntington’s Disease

Carsten Saft, Jürgen Andrich, Nina-Marie Meisel, Horst Przuntek and Thomas Müller [department of Neurology, St. Josef Hospital, Huntington center NRW, Ruhr University Bochum, Gudrunstrasse 56, 44791 Bochum, Germany

Introduction: Various complex instrumental tools and rating scales with respect to CAG repeat length may reflect severity of Huntington’s disease (HD). A simple assessment task for dysfunction due to the disease is the standardized, computer based performance of peg insertion. Objectives of our study were to compare scored HD symptoms, age related genetic disease load (CAG index = [CAG-35.5 x age]) and peg insertion scores between asymptomatic 34 HD gene carriers, 89 previously untreated HD patients with psychiatric or motor symptoms, or both, and 51 treated HD patients. We measured the period of the total time taken to insert 25 pegs from a rack into a series of appropriate holes, calculated the CAG index and scored the HD patients under blind conditions.

Results: Intervals for the peg insertion task significantly differed between HD patients and controls, but not between HD gene carriers and controls. CAG index reflected the increase of symptoms in HD. Peg insertion results and scored severity of HD and CAG index significantly correlated to each other. Conclusions: Peg insertion scores are no specific diagnostic marker for HD, but they reflect clinical symptoms of neurodegeneration. Performance of peg insertion involves visuospatial cognition, self-elaboration of internal strategies, sorting and planning of movement. Therefore peg insertion particularly reflects executive dysfunction in early HD and additionally motor impairment in advanced HD.

ABSTRACT 080-LATE-BREAKING RESEARCH: Is 41 better than 45? - Influence of low CAG ranges on Huntington´s disease

Saft C [department of psychiatry, WWU Muenster, Germany; department of neurology, St Josef Hospital, huntington center NRW, Ruhr-university Bochum, Germany], Andrich J, Brune N, Kraus PH, Przuntek H [department of neurology, St Josef Hospital, huntington center NRW, Ruhr-university Bochum, Germany], Gencik M, Epplen J [department of molecular genetics, huntington center NRW, Ruhr-university Bochum, Germany]

Age of onset of HD is known to be negatively correlated with an expanded CAG repeat in the HD gene. However, there is a broad mean variation especially in the lower CAG ranges. We investigated 154 patients (74 female, 80 male) previously analysed with CAG repeat numbers in a low range between 41 and 45 in order to study differences in main symptoms and age of onset. Age of onset of first motor and/or psychiatric symptoms was evaluated in every patient. 40 patients could be divided into groups of paternal or maternal transmission. Clinical assessment of symptoms was performed by using a standardised battery of neurological and psychological investigations. Clinical signs were differentiated as symptoms presenting or as main symptoms.

Mean age of onset of motor symptoms was 47.47 (±8.8) years, in psychiatric abnormalities (missing data for 33 patients) 45.87 (±9.0). Age of onset was 3.96 (motor), resp. 3.6 (psychiatric) years earlier than in parent if there was maternal transmission, but 4.84 (motor), resp. 6.0 (psychiatric) years earlier if there was paternal transmission. 97.4% of the patients showed symptoms of chorea, 51.3% suffered from choreatic movements as the main clinical symptom. About 29.9% (2.6% as main symptom) of the patients presented other movement disturbances like dystonia, tremor, myoklonus or akinesia. Major depression was presented by 64.3% (20.1% as main symptom) of all patients. 27.2% of the patients suffered from signs of a psychosis, in 7.1% of the patients psychosis with illusions and hallucinations was the main symptom. Dementia was found in 83.1% (18.8% as main symptom). Remarkably, we found a significant negative correlation in age of onset of HD for motor signs (r=-0.521, p=0.01), as well as for psychiatric abnormalities (r=-0.445, p=0.01) but no difference in the main symptoms in this genetically homogenous collective.

ABSTRACT 081-LATE-BREAKING RESEARCH: Emerging Cohort Characteristics in the Prospective Huntington At Risk Observational Study (PHAROS)

Huntington Study Group/PHAROS Investigators (presenter: Kevin M. Biglan, MD, University of Rochester; Rochester, NY)

Background & Objective: The Prospective Huntington At Risk Observational Study (PHAROS) was designed to determine the earliest and most specific signs of Huntington’s disease (HD) in clinically unaffected adults at risk for HD. As enrollment nears completion (final enrollment in 3rd quarter 2003), an analysis of the baseline characteristics of the PHAROS cohort was undertaken.

Design/Methods: Eligible research participants included unaffected adults (26 to 55 years old) at 50% risk for HD who agreed to prospective evaluations about every 9 months under conditions wherein individual HD gene status is never disclosed. Basic demographic and disease-specific data were obtained from standardized assessments at the initial (baseline) visit.

Results: As of May 1, 2003, 863 consenting adults have been enrolled at 43 PHAROS researchsites in North America and have been followed for 2.2 ± 1.0 years. The cohort, mean age =41.5 ± 7.5 years, is disproportionately female (69%), highly educated (mean education = 15 ±2.6 years), and largely employed (only 4% not in the labor force). The participants are married (71%) with a prevalence of divorce (13%) similar to the U.S. population as a whole (10%). At baseline, a small proportion (5%)of individuals reported symptoms possibly of HD, but this was not corroborated by the participants’ family members or by a significant difference in motor scores on the Unified Huntington Disease Rating Scale. A self-reported history of psychiatric illness was common (29% depression, 10% suicidal ideation, 4% suicide attempt, and 25% of participants were taking anti-depressants]. Depression occurred twice as commonly in women than men [33% and 17% respectively], but mean Beck depression scores were 2.2 ± 3.2 (mode 0 out of a maximum of 39) without gender disparity.

Conclusions: The cohort is more highly educated and more likely to be employed than the general population, consistent with the demographics of individuals who participate in clinical trials (J Chron Dis 29: 331-339, 1976), but perhaps not representative of the HD at-risk population as a whole. Despite the burdens and stress of HD, marital and divorce rates are similar to the general population.Research participants were more likely to report symptoms of HD and depression than could be verified thus far by independent measures.Ongoing longitudinal follow up of this cohort will enhance our understanding of the factors that may or may not relate to HD gene status and the development of signs of HD.

Financial Disclosure: The authors do not have any personal financial interest in this research.

This research is supported by the National Institutes of Health (HG 02449), the Hereditary Disease Foundation, the High Q Foundation, and the Huntington’s Disease Society of America.

ABSTRACT 082: Implicit processes in people with Huntington’s disease

Kristy Bolter, John McDowall (Victoria University of Wellington)

Evidence from previous implicit (or non-conscious) learning research with patients with various neurodegenerative diseases, including Huntington’s disease (HD), suggests that the corticostriatal system, the cerebellum, and the primary motor and sensory cortices may be implicated in implicit learning deficits. Furthermore, many previous studies with HD patients have indicated that poor performance on such tasks is not associated with the severity of motor symptoms but rather with cognitive status, which suggests that motor control dysfunction cannot completely account for the deficits in motor-based learning and memory in HD. One of the primary implicit learning tasks is the serial reaction time (SRT) task. However, one of the factors complicating the interpretation of SRT task impairment in patients with basal ganglia disorders is that movement disorders result in slower initial performance and more performance variability. Although research that has used the standard button pressing SRT task with this patient group shows reasonably robust deficits, a verbal version of the SRT was used in Experiment 1 to help to determine which neural substrates underlie this type of implicit learning and whether or not the task reflects a general sequence learning impairment. Results indicated that HD patients in the mid-later stages of the disease demonstrate sequence learning deficits even when the motor component of the task was minimised. In contrast, most types of implicit memory appear to be intact for this patient group. Experiment 2 focused on implicit memory (or priming) but also investigated different types of explicit memory and was the first study to examine the distinction between perceptual and conceptual processing in these two areas of memory. The results indicated intact conceptual and perceptual priming for people with HD and suggested further evidence for difficulties in initiating retrieval strategies in the explicit memory tasks. Theoretical and practical implications will be discussed.

ABSTRACT 085: Episodic Memory Impairment in Huntington’s Disease: a Meta-Analysis.

Alonso Montoya M.D., Marc Pelletier B.A., Elisabeth Duplessis B.Sc., Martin Lepage Ph.D (Brain Imaging Group, Douglas Hospital Research Centre, McGill University. 6875 Boul. LaSalle Verdun, Québec H4H 1R3)

Background: Memory dysfunction is an important feature in the clinical presentation of Huntington’s Disease (HD). Despite some equivocal findings, there is a prevailing notion that patients with HD exhibit retrieval-based episodic memory impairment. This assumption is based on the fact that HD patients often exhibit greater memory impairments on tasks of free recall compared to tasks of recognition memory. Some studies, however, have reported significant recognition memory impairments. Thus, a quantitative review of the existing literature is needed to better characterize the episodic memory impairment observed in HD. Objective: To assess the presence and magnitude of recognition memory impairment in HD and to contrast it to the free recall impairment using a meta-analytical strategy.Methods: We conducted a meta-analysis of published studies of recall and recognition memory in HD. Pubmed and Psyclit search engines were used to identify studies published between 1985 and December 2002 that reported measures of recognition and free recall memory in HD. We included only studies that compared performance of a group of HD patients to a healthy control group, and provided sufficient data to calculate an effect size. Results: Thirty studies totalling 381 symptomatic gene carriers, 487 presymptomatic gene carriers (individuals carrying the HD gene but who are free of symptoms), and 855 controls subjects met our criteria. The overall group comparison between symptomatic gene carriers and controls yielded a mean weighted effect size of d=1.45 for free recall and d=1.49 for recognition (z = 0.06, p =n.s.). The group comparison for presymptomatic gene carriers yielded effect sizes of .39 and .29 for recall and recognition respectively, again suggesting a similar pattern of impairment in both clinical groups.Conclusion: These results suggest that contrary to what some studies have suggested, recall and recognition memory are similarly impaired in both symptomatic and presymptomatic HD patients.No personal financial relationship of the authors with any products or sponsors. Supported by the Huntington Society of Canada and by the Fonds de Recherche sur la Nature et les Technologies du Québec.

ABSTRACT 089-LATE-BREAKING RESEARCH: Initial Baseline Characteristics in the Predict-HD (Neurobiological Predictors of HD) Study

Huntington Study Group/Predict-HD Investigators (presenter: Jane S. Paulsen, The University of Iowa)

Background & Objective: Predict-HD is a NINDS-funded study designed to examine early neurobiological and neurobehavioral markers of disease in presymptomatic HD using measures from MRI and cognitive neuroscience.Design/Methods: Unaffected adults (at least 26 years old) who had previously undergone genetic testing for the HD mutation were considered eligible research participants. Standardized assessments at the initial (baseline) visit included a comprehensive touch-screen neuropsychological examination, behavioral rating scales and a MRI, as well as basic demographic and disease-specific data.Results: 150 consenting participants have been enrolled at 15 Predict-HD research sites in North America as of May 1, 2003. The cohort, mean age = 41.7 ± 9.9 years, is 58% female, well educated (mean education = 14.5 ± 2.44), and predominately employed (only 0.67% not in the labor force). The participants are married (70.67%) with a prevalence of divorce (11.6%) similar to the U.S. population as a whole (10%). At baseline, a small proportion (13.33%) of individuals reported having possible symptoms of HD, and an even smaller proportion of families reported seeing possible symptoms (10.67%). These reports were not corroborated by the motor scores on the Unified Huntington’s Disease Rating Scale. Psychiatric illness by self-report was more than expected (42.67% depression, 13.33% suicidal ideation, 6% suicide attempt, and 62% of participants were taking anti-depressants]. Depression was more common in women than men [51% and 30.65% respectively].Conclusions: Depression was over twice as common in the Predict sample than that reported in the general population. Suicide attempt was four to six times greater and suicidal ideation was eight to 13 times than reported in the general population (Farrer, 1986; NIMH, 2003). These increased rates of depression are consistent with previous research conducted in at-risk and diagnosed HD samples (Almquist et al., 1999; DiMaio et al., 1993). Although the underlying mechanisms of depression and suicide are only beginning to be elucidated, it is critical for health care providers to include depression and suicide risk assessment as part of the traditional medical evaluation for HD.

Financial Disclosure: The authors do not have any personal financial interest in this research. This research is supported by the National Institutes of Health (NS440068 and MH01579 to JS Paulsen), the Huntington’s Disease Society of America and the Huntington Study Group.

ABSTRACT 091: Pupillary measure of brainstem activation is reduced in Huntington’s disease

Nadera Djerroud, Sylvain Chouinard, Francois Richer, Centre Hospitalier de l’Université de Montréal

Huntington’s disease (HD) is associated with a dysfunction in frontostriatal brain systems but little is known on the effect of this dysfunction on brainstem activation systems. We examined this question by measuring pupillary responses in patients in the early stages of HD and controls during auditory reaction time tasks. Subjects had to make rapid keypress responses after hearing letters A or B presented as singletons (simple responses) or triplets (sequential responses) while pupil size was measured with a head-mounted infrared camera. Patients were slower and made more errors in sequential responses than controls. There was no difference between groups in the baseline pupillary amplitude and pupillary light reflex was strong in all subjects. The amplitude of the pupillary response in HD patients was smaller than that of controls in both tasks and it was also less responsive to task difficulty. These results suggest that HD affects the recruitment of brainstem activation by fronto-striatal systems in relation to task demands and they are consistent with data on cortical hypoactivation in HD.

TOPIC: NEUROIMAGING

ABSTRACT 030: Progression of structural neuropathology in Huntington’s Dease using voxel and tensor based morphometryChristopher M Kipps, Andrew J Duggins, Neil Mahant, Elizabeth McCusker (Dept Neurology, Westmead Hospital, Wentworthville, Sydney NSW 2145)

Reliable measurement of progression of pathology in early HD is essential for the development of therapies to delay the onset and slow the progression of clinical disability. In HD there is evidence that atrophy occurs well before the clinical diagnosis of disease onset. 30 subjects (17 preclinical gene positive, 13 gene negative) were assessed both clinically and with MRI scanning over two years. Statistically significant regional changes in grey and white matter volume were analysed using both voxel-based morphometry (VBM) and tensor-based morphometry (TBM) techniques. VBM allows identification of voxels at which there has been a significant change in the probability of grey (or white) matter between initial and follow-up scans within subject. In TBM a voxel-wise estimation of volume change between images is calculated. Regions of brain atrophy (or expansion) are thus defined. Over the two year interval, both analyses revealed significant areas of regional grey matter atrophy in preclinical gene positive relative to gene negative subjects. Clinical progression over the same period did not reach significance. Both VBM and TBM techniques identified significant grey matter volume loss over 2 years in the right putamen. Significant grey matter atrophy over time was also seen in the right head of caudate with VBM, and the left head of caudate and putamen with TBM. VBM alone revealed significant cortical atrophy over time in bilateral intra-parietal sulci, right inferior frontal sulcus and the left fusiform gyrus. This study demonstrates that over a relatively short time period in preclinical Huntington’s Disease, significant regional grey matter atrophy can be detected using these techniques. The significance of these findings for therapeutic trials in Huntington’s Disease is clear: VBM and TBM could be used to assess the efficacy of potential disease modifying drugs in slowing the progression of pathology before clinical disease onset becomes apparent.

ABSTRACT 056: Quantification of Caudate and Putamen atrophy by volumetric MRI in patients with Huntington Disease. Association between length of the CAG repeat and age at onset.Heloísa Helena Ruocco,Íscia Lopes-Cendes, Marilza Santos Silva, Li Li Min, Fernando Cendes. (Universidade Estadual de Campinas-UNICAMP), (Cidade Universitária Zeferino Vaz-Barão Geraldo-UNICAMP-Campinas-SP-Brasil.

The neurobiology of Huntington Disease (HD) involves selective neuronal cell death, which is most striking in the caudate and putamen. However, the association between severity of CNS atrophy and age at onset or length of the CAG repeat is controversial. The objectives are to evaluate the degree of caudate, putamen and brain volume atrophy, by volumetric MRI and to determine if these findings correlate with molecular characteristics of HD. We studied 40 patients with HD (20 women) with a mean age of 45 years (21 to 69). The patients were genotyped for the CAG repeat in the HD gene. MRI images were obtained in a 2T system. Manual delineation of caudate and putamen volumes and total intracranial volumes were performed on T1 coronal 3 mm acquisitions using the NIH image program. Volumes obtained were compared to a control group of 40 normal volunteers. Values below 2 standard deviations from the mean of the control group were considered abnormal. The mean age at onset of the disease was 35.5 years (12 to 59). The mean expanded CAG repeat was 54.5 CAGs (40 to 69). All HD patients presented marked atrophy of the caudate nuclei, putamen and cerebral volume in comparison to controls (p<0.0001). An inverse correlation was found between the length of the expanded CAG tract and two variables: age at onset of the disease (p=0.000001) and total brain volume (p=0.0001). However, we did not find a linear correlation between expanded CAG length and the degree of atrophy of the caudate nuclei (p=0.80) or putamen (p=0.81). Volumetric MRI identified caudate and putamen atrophy in all patients with HD. The length of the expanded CAG tract correlated with an earlier onset of HD and with total brain atrophy, but it did not correlate to the degree of atrophy in specific CNS structures.

ABSTRACT 059: Brain Energy Metabolism in Huntington’s disease

Marguerite Wieler, Chris Hanstock, Wayne Martin (University of Alberta, Edmonton, Alberta, Canada)

Mitochondrial dysfunction may play a role in the pathogenesis of Huntington’s disease (HD). Several investigators have reported an increase in brain lactate content in HD using proton magnetic resonance spectroscopy (MRS), implying impaired mitochondrial function. These observations have been challenged, however, with the suggestion that the reported increase in brain lactate may reflect contamination from ventricular cerebrospinal fluid (CSF) within the voxel of interest. The objective of the present study was to evaluate the potential contribution of CSF lactate to the changes observed with single voxel MRS in HD. We utilised MRS at 3.0 T to examine occipital lactate levels in 8 HD patients [age 47 yrs. (38-61 yrs.); symptom duration 4.8 yrs.(1-14 yrs.); UHDRS motor scores 31.3 (15-48)] and in 11 normal controls [age 46 yrs. (35-67 yrs.)]. We segmented voxels into brain and CSF, determining the percentage of each in the voxel of interest. Spectroscopic data were expressed as the ratio of lactate to the neuronal marker N-acetyl aspartate (lac/NAA). In voxels ranging from 6% - 78% CSF, there was a close correlation between lac/NAA and the amount of CSF in both groups (HD: r = 0.99; controls: r = 0.97). By extrapolating lac/NAA to a voxel containing 100% brain, we determined this ratio to be substantially greater in HD vs controls, suggesting increased lactate content in this predicted voxel. We conclude that although MRS does indicate increased brain lactate in HD, results are highly dependent on voxel placement and resulting variations in CSF content in the voxel of interest.

ABSTRACT 065: MRI and PET assessment of brain involvement in subjects with presymptomatic, initial and advanced Huntington disease.

Andrea Ciarmiello, Secondo Lastoria (Istituto Nazionale dei Tumori IRCCS Pascale, Via Mariano Semmola -80131- Naples Italy) Ferdinando Squitieri, Milena Cannella (Neurogenetics Unit IRCCS Neuromed Località Camerelle 86077 Pozzilli -IS- Italy)

Study aim was to define metabolic and morphologic brain changes using [(18)F]2-fluoro-2-desoxy-D-glucose (FDG) Positron Emission Tomography (PET) and Magnetic Resonance Imaging (MRI) in patients with Hungtinton disease (HD). Forthy-eight subjects (24 males, 24 females, age range 25-70) from presymptamtic to advanced HD stages, were enrolled into the study. A data-base of 53 normal subjects MRI studies (NV), has been used as control group and to assess age-related changes in brain tissues. Static [(18)F]FDG-PET scans was performed in all patients and in a group of 10 gene negative subjects with an ECAT EXACT HR (CTI/Siemens, Knxville, TN). MRI scan was performed using T1w (600/15 TR/TE) and DP/T2w (2200/15-90 TR/TE1-TE2) conventional spin-echo images, covering the entire brain. Segmentation of MR brain images into gray matter (GM), white matter (WM) and cerebrospinal fluid (CSF) was obtained using a fully automated multispectral approach (Alfano, 1997). For each study, total GM, WM and CSF were divided by total intracranial volume (ICV), thus providing fractional results (fGM, fWM, and fCSF). MR segmented images and PET scans were evaluated by using SPM99 (Wellcome Department of Cognitive Neurology, London). Images were spatially normalized to costumized template in a standard stereotactic space and smoothed with an isotropic Gaussian Kernel of 12-mm full with half maximum (FWHM). Significant decrease in fGM was present in HD patients as compared to NV (46.95±3.5% vs. 53±2.8%; p<0.0001), with a corresponding increase in fCSF (22.27±6.1% vs. 9.3±3.3%; p<0.0001). Furthermore a significant decrease was also found in fWM (30.52±3.5 vs 37.67±2.21; p<0.0001). Fractional GM a WM showed a significant correlation to the HD stage, disability and progression rate. SPM analysis showed significant reduction of cerebral glucose metabolism mainly involving basal ganglia and frontoparietal cortex bilaterally. Gene-positive (n=3) pre-simptomatic subjects showed involment of frontoparietal cortex with no evidence of F DG uptake reduction in basal ganglia.

ABSTRACT 084-LATE-BREAKING RESEARCH: Onset and Rate of Change in Basal Ganglia Volume in Presymptomatic Subjects

Elizabeth H. Aylward,Bobbi-Faun Sparks, Katherine M. Field, Venugopal Yallapragada (University of Washington) Barnett D. Shpritz, Adam Rosenblatt, Jason Brandt, Lisa M. Gourley, Russell L. Margolis, Christopher A. Ross (Johns Hopkins University School of Medicine) Kung-Yee Liang,Hongling Zhou (Johns Hopkins University School of Public Health)

Two studies, one cross-sectional and one longitudinal, were designed to address three questions: (1) When does basal ganglia atrophy begin in the presymptomatic stage? (2) What is the rate of atrophy prior to onset? (3) Can basal ganglia volumes predict onset of diagnosable HD? The cross-sectional study involved measures of caudate and putamen volume on MRI scans from 19 subjects who were very far (9-20 years) from estimated onset and controls who were matched on age and sex. The longitudinal study included 17 subjects who had two or more MRI scans (mean = 4.1 scans), covering a period of 1 to 6 years (mean interval = 4.2 years). All subjects were presymptomatic at the initial scan, but six subjects in the longitudinal study were diagnosed with HD sometime during the course of the study. Results indicate that even those subjects who were very far from onset had significantly smaller striatal volumes than controls. General Estimating Equations analysis indicated that significant longitudinal change begins approximately 11 years from estimated onset in caudate and approximately 9 years from estimated onset in putamen. Rates of change for subjects in the longitudinal study were 3.5% per year for caudate and 2.7% per year for putamen. For the six incident cases, caudate volume was about ½ that of controls at the time of diagnosis; putamen was 1/3 to ½ of normal volume. A discriminant function analysis indicated that caudate volume predicted with 100% accuracy those subjects who would become symptomatic within 2 years of imaging. We conclude that basal ganglia volumes, as measured on serial MRI scans, may be excellent outcome measures for clinical trials in presymptomatic subjects with the HD gene mutation. Our research was supported by: NINDS (NS16375), the Johns Hopkins School of Medicine General Clinical Research Center (NIH/NCRR grant M01 RR00052), and the Huntington’s Disease Society of America. None of us has any personal financial relationship with this organization or any other manufacturer of commercial paroducts to be discussed.

ABSTRACT 090: Frontal hypoactivation and reduced voluntary control in early Huntington’s disease

Chantal Lafrance, Sylvain Chouinard, Francois Richer, Centre Hospitalier de l’Université de Montréal

Huntington’s disease affects the voluntary control of movements. The present study examined the changes in brain activity linked to this control problem using functional magnetic resonance imaging (fMRI) during a visuomotor control task. Seven patients with early HD and nine controls had to trace circles with a joystick while monitoring their movements on a screen in 3 conditions: a) moving inside a ring of 3 mm-depth, b) moving inside a ring of 15 mm-depth and c) with no ring boundaries. Blood oxygenation fMRI images were acquired on a 1.5 T MRI system and 80 measurements were obtained for each condition in a block design. Trajectory precision was reduced in patients. Regional activation changes were assessed in terms of t-values (p<0.05, corrected for small volumes). The HD group showed reduced activation in ventral frontal cortex, thalamus and striatum in all conditions. Comparisons between high-precision and low-precision conditions showed enhanced activation in lateral and medial frontal cortex in controls but patients showed a reduced modulation of activity with task demands. The data suggest that both general hypoactivation and lowered task-related modulation of cortical activity can be observed in early HD.

TOPIC: HUMAN EXPERIMENTAL THERAPEUTICS

ABSTRACT 001: Targeted nucleotide exchange in the CAG repeat region of the human HD gene

Hetal Parekh-Olmedo, Yiling Hu, Eric B. Kmiec (University of Delaware, Delaware Biotechnology Institute; 15 Innovation Way; Newark, DE 19711)

Huntington’s Disease (HD) is marked by the expansion of a tract of repeated CAG codons in the HD-gene, IT15. Once expressed, the expanded poly Q region of the huntingtin protein (Htt), which is normally soluble, becomes insoluble, leading to the formation of intracellular inclusions and ultimately to neuronal degeneration. Interruption of the pure poly Q tract at the genetic level should undermine the transition from Htt solubility to Htt insolubility. Modified single-stranded oligonucleotides were used to direct the nucleotide exchange of an A residue to a T residue in the second codon of the HD-gene polyQ repeat, resulting in the creation of a leucine residue among the poly Q tract. We have also created TAG step codons at several positions within the same CAG gene tract. Consistent with results from other groups, we provide evidence that short synthetic DNA molecules can modify the HD-gene directly, preliminarily offering a potential therapeutic approach to Huntington’s Disease.

ABSTRACT 022: Increased Cell Proliferation and Neurogenesis in Huntington’s Disease

Maurice A Curtis 1, Ellen B Penney 1, John F Pearson 2, Michael Dragunow 3, Bronwen Connor 3, Richard LM Faull 1. (Departments of Anatomy with Radiology 1, Statistics 2, Pharmacology 3.) (The University of Auckland. Auckland, New Zealand)

The recent demonstration of endogenous stem/progenitor cells in the adult mammalian brain raises the exciting possibility that these undifferentiated cells may be able to generate new neurons for cell replacement in neurodegenerative diseases such as Huntington’s disease (HD). Neural stem cells in the rodent brain subependymal layer (SEL), adjacent to the caudate nucleus, proliferate and differentiate into neurons and glial cells1 suggesting they may provide a source of replacement neurons. Previous studies on the normal adult human brain have shown evidence of neurogenesis in the hippocampus but no previous study has shown neurogenesis in the diseased human brain. Using an antibody to the mitotic marker proliferating cell nuclear antigen (PCNA), we have observed increased numbers of PCNA immunopositive cells in the SEL adjacent to the caudate nucleus in HD (n=9) compared to control (n=6) human brains. The increase in number of PCNA positive cells correlates (p<0.003) with increased pathology and CAG repeat length (p<0.02). Furthermore, bIII tubulin and PCNA colocalised in a subset of cells in the SEL indicating neurogenesis in the HD brain. We also serially sectioned and stained the entire striata of 3 control and 3 HD (grade2/3) brains for PCNA and completed PCNA cell counts throughout the SEL region. Statistical analysis showed a 4-fold increase in the number of PCNA positive cells in HD brains compared with controls; most notably the ventral SEL contained the highest number of cells. This is the first study to map the pattern of stem/progenitor cell proliferation in the SEL adjacent to the caudate nucleus of the basal ganglia and provides new evidence of the pattern of neurogenesis in the human brain.

1. Morsehead, C. et al. Neuron (13) 1071 (1994)

Supported by: Health Research Council and Neurological Foundation of NZ.

The authors have no personal financial relationships.

ABSTRACT 032: Treatment of Huntington’s Disease Using Essential Fatty Acids (long term follow up case presentation)

Dr. Krishna S. Vaddadi, Associate Professor of Psychiatry, Monash University and Monash Medical Centre, 246 Clayton Road, Clayton 3168, Victoria, Australia. Dr. Philip Dingjan, Senior Clinical Neuropsychologist, Department of Psychiatry, The University of Melbourne and St. George’s Health Service, 238 Cotham Road, Kew, 3103, Victoria, Australia. Ms Eileen A. Soosai, Research Nurse, Monash Medical Centre, 246 Clayton Road, Clayton 3168, Victoria, Australia.

Huntington’s disease is a lethal, dominantly inherited neurodegenerative disease caused by CAG expansion of a poly-CAG section beyond 35 repeats in HD gene. The brain is unique in high concentration of long chain, highly unsaturated fatty acids (HUFA) which play an important role in the structure and function of brain tissue. Highly unsaturated fatty acids have been implicated in a number of neurological conditions from Alzheimer’s disease to Huntington’s disease. Optimal supply of HUFA, coupled with effective antioxidant system would be essential for neuronal survival, function and prevention of apoptosis. Open case studies, transgenic mouse model of Huntington’s disease and a double blind clinical trial using HUFA containing EPA are encouraging in ameliorating motor abnormality in Huntington’s disease. We have used essential fatty acids containing Ethyl-eicosapentaenoic acid (EPA) in treating patients with Huntington’s disease. A single treatment case example will be presented, including video presentation for eight years follow up. Cognitive measures have been shown to be amongst the more sensitive clinical indicators of progression in Huntington’s neuropathology and correlate well with structural and functioning neuro-imaging. For this treatment case example, serial neuropsychological assessment data over a six year treatment period show limited variation in scores but not progressive deterioration in measures of executive and new verbal learning ability. At the last assessment, all test scores were within one standard deviation of baseline values. The lack of deterioration across such a long time scale, and with a 20 month gap between the last and the penultimate assessment times, suggests that treatment with HUFA therapy has been clinically beneficial. A theoretical review of the essential fatty acid therapy in Huntington’s disease will be presented.

ABSTRACT 049: Measurement of Irritability in Huntington’s Disease

David Craufurd, Laura Smyth (University of Manchester) Elizabeth Howard (St. Mary’s Hospital)Jennifer Thompson, Julie S Snowden (Greater Manchester Neuroscience Centre, Manchester, UK)

Irritability and angry outbursts are common in patients with Huntington’s disease (HD) and often have a major impact on the clinical care and quality of life of both patients and families. In a recent study of 134 patients attending an HD management clinic, 69 (51.4%) were irritable during the month prior to assessment, while 53 (39.5%) had verbal outbursts and 29 (21.7%) displayed physical aggression. Clinical experience suggests that these symptoms can be ameliorated with currently available psychotropic medications, but there is no published evidence base and clinical trials are needed to demonstrate the efficacy of agents such as beta blockers and specific serotonin reuptake inhibitors. In the longer term, trials of novel therapies for HD will need to demonstrate their efficacy with respect to major behavioural symptoms such as irritability as well as the motor and cognitive aspects of the condition.Most existing measures of irritability and aggression are unsuitable for use with HD patients. Self-rating scales are limited by the problems of denial and cognitive impairment, while standardised psychiatric interview schedules overlook irritability because it is not a core symptom of most psychiatric disorders. We conducted a pilot study using the Overt Aggression Scale (OAS) (Yudofsky et al., 1986), which requires an observer rating for each angry outburst, but carers found this burdensome and frequently reported that the subject would have become angry if they had not behaved with great tact, or kept out of the way. We therefore report the development and validation of a new measure which detects irritability (the propensity to become angry with relatively little provocation) as well as actual aggression, is sensitive to minor changes in the severity of angry behaviours, and may be more suitable for use in clinical trials with HD patients. No personal financial relationship to product or sponsor.

ABSTRACT 063: Ethyl-eicosapentaenoate (ethyl-epa) in Huntington’s disease: a randomized, placebo-controlled trial

Blair R. Leavitt and Michael R. Hayden (University of British Columbia, Vancouver, BC, Canada)Christopher A. Ross and Adam Rosenblatt (Johns Hopkins Medical School, Baltimore, MD, USA)Basant K. Puri (Hammersmith Hospital, London, United Kingdom), J. Timothy Greenamyre (Emory Medical School, Atlanta, GA, USA), Steven M. Hersch (Harvard Medical School, Charlestown, MA, USA), Krishna S. Vaddadi (Monash Medical Centre, Clayton, Australia), Alastair Sword and David F. Horrobin (Laxdale Ltd, Stirling, United Kingdom)

This trial was designed to test the effect of Ethyl-EPA versus placebo on motor signs in HD. 135 patients with early stage HD entered a double-blind study at three US centers and one center each in Canada, the UK and Australia. Patients were randomized to receive 2g/d Ethyl-EPA or matching placebo for one year. Ethyl-EPA was generally well tolerated with similar adverse events to placebo. The primary end-point of the trial was the total motor score-4 (TMS4) sub-scale of the Unified Huntington’s Disease Rating Scale (UHDRS). Secondary end-points were other UHDRS sub-scales and the total UHDRS. Pre-planned analyses were carried out on both an intent-to-treat (ITT) group that included all 135 randomized patients, and a per protocol (PP) group that excluded 38 patients for non-compliance with protocol. Pre-planned analyses on both groups were: ANCOVA with center, baseline TMS4 (disease severity), CAG size and treatment as covariates; and chi square with a binary division into patients who improved or deteriorated from baseline. On the primary ITT analysis, there was no significant difference between Ethyl-EPA and placebo. In the PP analysis, Ethyl-EPA was significantly better than placebo on chi square (p<0.05) but just missed significance on ANCOVA (p=0.06). Because non-compliant patients tended to be more severely affected, the PP group was significantly less symptomatic at baseline than the ITT group. No significant effects of Ethyl-EPA were seen on total UHDRS or other sub-scales. Sub-group analysis revealed no effect of center, but baseline TMS4 score and CAG size appeared to significantly influence outcome in both the ITT and PP Groups. Ethyl-EPA may have a modest beneficial effect in HD. Further trials are indicated to confirm these findings, and determine whether the effect is immediate or represents a slowing of progression.

ABSTRACT 067: Management of Huntington’s disease: An evidence-based review

Raphael M Bonelli, MD, Theresa Lahousen, MD, (University Clinic of Psychiatry, Graz, Austria)Klaus Seppi, MD, Gregor K Wenning, MD, PhD (University Clinic of Neurology, Innsbruck, Austria)

Huntington’s disease (HD) is a relentlessly progressive, neurodegenerative disorder with currently no cures or even effective therapies. Indeed, there is no level A evidence of the management of HD available today. We analysed all available double-blind, placebo controlled studies on therapy in HD (level B evidence). Moreover, we included all relevant open trials and case reports. Interestingly, the only existing controlled trials are focused on the treatment of chorea, while other important features of HD like dementia, depression, psychosis were widely ignored in the past. We recommend riluzole, olanzapine, and amantadine for the treatment of the movement disorders associated with HD, SSRI and mirtazapine for the treatment of depression, atypical antipsychotic drugs for HD psychosis and behavioural problems. Moreover, adjuvant psychotherapy, physiotherapy, and speech therapy should be applied to supply the optimal management.

ABSTRACT 088: Mechanism of Action of the Anti-Chorea Drug tetrabenazine: A Review

1: Quinn et al, 1959; Butcher and Anden, 1969; Bagchi, 1983; 2: Pettibone et al, 1984; 3: Scherman et al, 1981; 1983; 1986; 4: Scherman et al, 1988; 5: Koeppe et al, 1997; Gilman et al, 1996; Bohnen et al, 2000 6: Gonzalez et al, 1994; 7: Schuldiner et al, 1993; Kilbourn et al, 1995; Erickson et al, 1996; 8: Scherman and Henry, 1984; Darchen et al, 1989.

Tetrabenazine is a potent and selective depletor of monoamines from nerve terminals.1 Within the CNS, tetrabenazine causes preferential depletion of dopamine2 (DA). The IC50 for DA depletion is approximately 0.4 mg/kg, whereas the IC50 for norepinephrine (NE) and 5-HT depletion is approximately 2 mg/kg. The CNS-selective and DA-preferential effects of tetrabenazine are linked to its molecular mechanism of action. Although tetrabenazine has been successfully used for the treatment of chorea for over 40 years, its exact molecular mechanism of action has only been recently fully understood. Tetrabenazine inhibits monoamine uptake into synaptic vesicles by binding with high affinity (Kd = 2.4 nM) and selectivity to the principal brain synaptic vesicular monoamine transporter, VMAT.3 Binding is reversible with a half-lifetime of dissociation of about 15 minutes. In humans, post-mortem binding studies show that the highest density of binding sites occurs in the basal ganglia.4 PET-scan studies in healthy volunteers and in patients with Huntington’s disease, confirm that tetrabenazine binds mainly to striatal DAergic terminals.5 The gene for VMAT was cloned in the early 1990s, revealing the existence of two distinct, but highly related genes: VMAT1 and VMAT2. In humans, VMAT1 expression is excluded from the brain, and VMAT2 expression is nearly exclusively neuronal.6 Tetrabenazine exhibits a 10,000-fold higher affinity for human (h)VMAT2 than for hVMAT1.7 The high selectivity of tetrabenazine for hVMAT2 explains the CNS-selective effects of the drug. Reserpine, like tetrabenazine, depletes monoamines by binding to VMAT, however, reserpine binds to both VMAT1 and VMAT2.8 Unlike reserpine, tetrabenazine does not affect blood pressure or gut motility, suggesting that both effects involve VMAT1.

1: Quinn et al, 1959; Butcher and Anden, 1969; Bagchi, 1983; 2: Pettibone et al, 1984; 3: Scherman et al, 1981; 1983; 1986; 4: Scherman et al, 1988; 5: Koeppe et al, 1997; Gilman et al, 1996; Bohnen et al, 2000 6: Gonzalez et al, 1994; 7: Schuldiner et al, 1993; Kilbourn et al, 1995; Erickson et al, 1996; 8: Scherman and Henry, 1984; Darchen et al, 1989.

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