Understanding RRPa genetic undertaking

Allegheny General Hospital

Pittsburgh, PA

Center for Genomic Sciences

Allegheny Singer Research Institute

Farrel J. Buchinsky, MD

Pediatric Otolaryngologist

BLUF (bottom line up front)

Enrollment tripled since Los Angeles focus meeting

Discovered transmission disequilibrium in 2 candidate genes

What is the cause of RRP?

Necessary Is it sufficient?

Many exposed Only few get the disease

Genetic Susceptibility higher prevalence in relations

But still very rare HLA DRB1*0301 and DQB1*0201

disproportionately present in RRP associated with decreased interferon γ expression Partial reproduc of Gelder Also assoc in cervical cancer

Epidermodysplasia verruciformis Bonagura; Hum Immunol. 2004

Gelder; J. of Vir. 2003Madeleine; JID. 2002

Ramoz; Nature G. 2002

Epidermodysplasia Verruciformis Rare, autosomal recessive, skin disease Abnormal susceptibility to specific HPVs (5 and 8)

persistent lesions that do not clear virus 4 specific mutations found in EVER1

Dr. Mark Naylor, Electronic Textbook of Dermatology

More Genetic Evidence Rabbit papilloma data HIV/AIDS

14 AIDS restriction genes explains some of variability rate at which a population progresses from HIV infection to AIDS

Malaria, Otitis Media Mortality by infectious disease in adoptees

more associated with biologic parents than seen for cardiovascular and cancer

O'Brien; Nature G. 2004Sørensen; NEJM. 1988

From who?

So what’s needed

Specimen of Blood 2 specimens 1 mm * 1 mm

ORHPV type

Acquisition

Mouthwash specimens

No HospitalsNo Doctors

Enrolment of people with RRP and their

parentsSatellite Design

Support-Group Design

Papilloma and Blood Specimens

Coordinating CenterDr. Farrel Buchinsky

Ms. Marilyn SmithCenter for Genomic SciencesAllegheny-Singer Research

Institute

IRB

IRB

IRB

Enrolment

92 from Patient Support Groups

Recruitment SourceCount Institution City PI

92 Allegheny Singer Research Institute Pittsburgh Buchinsky

23 Children's Medical Center of Dallas Dallas McClay

21 Cincinnati Children's Hospital Medical Center Cincinnati Myer

17 The Hospital for Sick Children Toronto Campisi

14 Children's Hospital of Alabama Birmingham Wiatrak

12 Children's Hospital of Wisconsin Milwaukee Conley

11 Children's Hospital of the King's Daughters Norfolk Derkay

11 Children's Healthcare of Atlanta at Egleston Atlanta Sobol

11 Children's National Medical Center Washington Choi

10 University of Mississippi Medical Center Jackson Schweinfurth

7 Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo Sao Paulo Tsuji

5 SUNY Upstate Medical Center Syracuse Mortelliti

4 Johns Hopkins Children's Center Baltimore Tunkel

4 Le Bonheur Children's Medical Center Memphis Thompson

4 Children's Hospital of Philadelphia Philadelphia Elden

3 Children's Hospital of Pittsburgh Pittsburgh Dohar

1 University of Pittsburgh Medical Center Pittsburgh Rosen

1 Allegheny General Hospital Pittsburgh Buchinsky

1 British Columbia's Children's Hospital Vancouver Kozak

Which gene or genes 3x109 bases Candidate Gene Approach Genomic Scan Approach

Candidates MHC Innate Immunity

Toll-like receptors Cytokines regulating the inflammation Transcription factors regulating immune response

Known Cell Biology Interactions E5 inhibits H+ATPase from acidifying vesicles HPV cell receptor

Other Diseases Epidermodysplasia Verruciformis Cervical Cancer

First Candidate Gene Batching 5 genes 214 SNPs on Sequenom MassARRAY 212 cases + relatives

Transmission Disequilibrium Test

Mother Father

C DBA

C DA B

Result of all possible zygotes

A C B C A D B D

1/1 1/1

1/1

10

1/1 2/1

1/1

1

1/1 2/1

2/1

6

2/2 1/1

2/1

2

2/1 2/1

2/2

3

2/2 2/1

2/2

2

1/1 2/1

2/1

2

Allele 1 = 76%, Allele 2 = 24%

Many trios uninformative Allele 2 transmitted 16

times, untransmitted 1

TDTae in 2 candidate genes

Gordon; Euro J. Hum. Gen. 2004

Quality control scrutiny Additional SNP genotyping Sequencing

Whole Genome

96 humans at a time 300000 SNPs at a

time

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illumiCode #561

illumiCode #217

illumiCode #1024

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A/A T/T A/T

Acknowledgments CGS

Garth Ehrlich, Chris Post Joseph Donfack, Marilyn

Smith, Fen Hu

Rockefeller University Jurg Ott, Sara Hamon

Earliest Collaborators Craig Derkay, RRP Task

Force

Patient Support Groups Michael Green, Bill Stern

Funding NIH – Mentored Clinical

Scientist Development Award (Mentored Clinical Scientist Development Award (K08))

RRP ISA – IRB assistant

fjbuch@gmail.com

(412) 567-7870