Understanding RRP a genetic undertaking Farrel J Buchinsky

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) o o Enrollment tripled since Los Angeles focus meeting Discovered transmission disequilibrium in 2 candidate genes

What is the cause of RRP? Necessary Is it sufficient? o o

Many exposed o Only few get the disease

Genetic Susceptibility o higher prevalence in relations n o HLA DRB 1*0301 and DQB 1*0201 disproportionately present in RRP n n n o 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 o o Rare, autosomal recessive, skin disease Abnormal susceptibility to specific HPVs (5 and 8) n o persistent lesions that do not clear virus 4 specific mutations found in EVER 1 Dr. Mark Naylor, Electronic Textbook of Dermatology

More Genetic Evidence o o Rabbit papilloma data HIV/AIDS n o o 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

Acquisition IRB IRB

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 Mc. Clay 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 Elden 3 Children's Hospital of 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 o o o 3 x 109 bases Candidate Gene Approach Genomic Scan Approach

Candidates o o MHC Innate Immunity n n n o Known Cell Biology Interactions n n o Toll-like receptors Cytokines regulating the inflammation Transcription factors regulating immune response E 5 inhibits H+ATPase from acidifying vesicles HPV cell receptor Other Diseases n n Epidermodysplasia Verruciformis Cervical Cancer

First Candidate Gene Batching o o o 5 genes 214 SNPs on Sequenom Mass. ARRAY 212 cases + relatives

Transmission Disequilibrium Test o o o Allele 1 = 76%, Allele 2 = 24% Many trios uninformative Allele 2 transmitted 16 times, untransmitted 1

TDTae in 2 candidate genes o o o Quality control scrutiny Additional SNP genotyping Sequencing Gordon; Euro J. Hum. Gen. 2004

Whole Genome o A/A illumi. Code #217 #1024 //// illumi. Code #561 96 humans at a time 300000 SNPs at a time T/T //// o A/T

Acknowledgments o CGS n n o Jurg Ott, Sara Hamon Earliest Collaborators n o Funding n Rockefeller University n o Garth Ehrlich, Chris Post Joseph Donfack, Marilyn Smith, Fen Hu o Craig Derkay, RRP Task Force Patient Support Groups n Michael Green, Bill Stern n NIH – Mentored Clinical Scientist Development Award (K 08)) RRP ISA – IRB assistant fjbuch@gmail. com (412) 567 -7870