Download Understanding RRP: a genetic undertaking

Understanding RRP
a genetic undertaking
Farrel J. Buchinsky, MD
Pediatric Otolaryngologist
Allegheny General Hospital
Center for Genomic Sciences
Pittsburgh, PA
Allegheny Singer Research Institute
BLUF (bottom line up front)
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Enrollment tripled since Los Angeles focus
meeting
Discovered transmission disequilibrium in 2
candidate genes
What is the cause of RRP?
Necessary
Is it sufficient?
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Many exposed
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Only few get the disease
Genetic Susceptibility
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higher prevalence in relations
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HLA DRB1*0301 and DQB1*0201
disproportionately present in RRP
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But still very rare
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. 2003
Madeleine; JID. 2002
Ramoz; Nature G. 2002
Epidermodysplasia Verruciformis
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Rare, autosomal recessive, skin disease
Abnormal susceptibility to specific HPVs (5 and 8)
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persistent lesions that do not clear virus
4 specific mutations found in EVER1
Dr. Mark Naylor, Electronic Textbook of Dermatology
More Genetic Evidence
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Rabbit papilloma data
HIV/AIDS
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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. 2004
Sørensen; NEJM. 1988
From who?
So what’s needed
Specimen of Blood
2 specimens
1 mm * 1 mm
OR
HPV type
Enrolment of people
with RRP and their
parents
Acquisition
Satellite
Design
SupportGroup
Design
IRB
IRB
No Hospitals
No Doctors
IRB
Coordinating Center
Papilloma and Blood
Specimens
Dr. Farrel Buchinsky
Ms. Marilyn Smith
Center for Genomic Sciences
Allegheny-Singer Research
Institute
Mouthwash
specimens
Enrolment
92 from Patient Support Groups
Recruitment Source
Count
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
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3x109 bases
Candidate Gene Approach
Genomic Scan Approach
Candidates
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MHC
Innate Immunity
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Known Cell Biology Interactions
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Toll-like receptors
Cytokines regulating the inflammation
Transcription factors regulating immune response
E5 inhibits H+ATPase from acidifying vesicles
HPV cell receptor
Other Diseases
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Epidermodysplasia Verruciformis
Cervical Cancer
First Candidate Gene Batching
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5 genes
214 SNPs on Sequenom MassARRAY
212 cases + relatives
Transmission Disequilibrium Test
1/1
1/1
1/1
2/1
1/1
10
A C
1/1
B C
Result of all possible zygotes
1/1
2/1
2/1
2/2
2/1
2/1
2/2
2/2
1
2
6
2
3
2
C
A D
2/2
2/1
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Father
B
2/1
2/1
C D
Mother
2/1
1/1
A B
A
1/1
D
B D
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Allele 1 = 76%, Allele 2
= 24%
Many trios uninformative
Allele 2 transmitted 16
times, untransmitted 1
TDTae in 2 candidate genes
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Quality control scrutiny
Additional SNP genotyping
Sequencing
Gordon; Euro J. Hum. Gen. 2004
Whole Genome
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A/A
illumiCode illumiCode
#217
#1024
/\/\/\/
/\/\/\/
illumiCode
#561
96 humans at a time
300000 SNPs at a
time
T/T
A/T
/\/\/\/
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Acknowledgments
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CGS
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Jurg Ott, Sara Hamon
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NIH – Mentored Clinical
Scientist Development
Award (Mentored Clinical
Scientist Development
Award (K08))
RRP ISA – IRB assistant
Earliest Collaborators
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Funding
Rockefeller University
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Garth Ehrlich, Chris Post
Joseph Donfack, Marilyn
Smith, Fen Hu
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Craig Derkay, RRP Task
Force
Patient Support Groups
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Michael Green, Bill Stern
[email protected]
(412) 567-7870