understanding rrp a genetic undertaking allegheny general hospital pittsburgh, pa center for genomic...
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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
(412) 567-7870