apbdrf scientific advisory board meeting december 2013: research update
TRANSCRIPT
2013 Research Update
APBDRFH. Orhan AKMAN, PhD
Dr. DiMauro’s Laboratory
Columbia University Medical Center
Department of Neurology
New York
Adult Polyglucosan Body DiseaseDiagnosis, and Development of tools for Research and Treatment
Mouse model of APBD
Possible Pharmacological Treatments of APBD
Diagnosis of Manifesting Heterozygote Patients
Mouse model of APBD with Y329S mutation
Y329S TAT to TCTTyr Ser
FRT
LoxP
5’ ARM 2.1 kb PGK- Neomycin 3’ARM 6.5 kb Exon 7
Exon 75’ ARM 3’ARM
Generation of mouse model of APBD:
Breeding scheme and expected genotypes
Y329S TAT to TCTTyr Ser
FRT
LoxP
5’ ARM 2.1 kb PGK-Neomycin 3’ARM 6.5 kb Exon 7
Exon 75’ ARM 3’ARM
1 2 1
Expected & other genotypes
5’ PGK-Neomycin 3’Exon 7
5’ PGK-Neomycin 3’Exon 7
Y329S-Neo
WT-Neo
Exon 75’ 3’
Y329S
Exon 75’ 3’
WT-lox
5’ 3’
Exon-7 del or D
Difference between Gbe1Y329S and Gbe1neo/neo mice
Gbe1+/+ Gbe1neo/neoPercent
changeGbe1ys/ys
Percent
change
Brain 230 ± 17 25 ± 8 11% 41± 9 n=4 17.8%
Heart 457 ± 56 72 ± 34 16% 92± 21 n=4 20.1%
Liver 855 ± 43 115 ± 42 13% 210± 35 n=4 24.6%
Muscle 251 ± 23 38 ± 18 15% ND n=4 ND
High throughput screening
We have tested 1700 FDA approved compounds.
Analysis has yield 15 compounds decreasing PGB content in tissue culture
Validation of 15 HTS hits
0.0
1.0
2.0
3.0
4.0
5.01 2 3 4 5 6 7 8 9
10
11
12
13
14
15
Co
ntr
ol
Re
lati
ve G
lyco
gen
Co
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nt
(mic
rogr
am/m
g p
rote
in)
Diagnosis of APBD
GBE enzyme assay
Histologic examination of sural nerve biopsy
Sequencing genomic DNA and messenger RNA.
Search for the genetic change that eliminates the normal allele
1 to 6 5 to 10 9 to 14
12 to 15 14 to 16
C P1 P2 P3 C
C P1 P2 P3 C
THANK YOUMichio Hirano, MD Sergey Pompou, MDValentina Emmanuelle, MD Carmen Paradas, MD
Salvatore DiMauro, MDSindu Krishna, PhDPablo Abreu