1. Pediatrics Genome Wide Peripheral Blood Leukocyte DNA Methylation Microarrays Identify a Single Association with Inflammatory Bowel Diseases Richard Kellermayer

2. Page 2 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Etiology of IBD Intestinal mucosa Sartor RB. Nat Clin Pract Gastroenterol Hepatol. 2006

3. Page 3 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Kellermayer R. AJMG 2010 Systems of Key Elements in IBD Pathogenesis Kellermayer R. AJMG 2010

4. Page 4 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Increasing prevalence of IBDs argues against primary role of genes Molodecky NA et al. Gastroenterology 2011 Before 1960 1960-1980 1980-2008

5. Page 5 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Genetic contribution to IBD 40% explained by HLA locus 71 susceptibility loci (Franke A et al. Nat Genet 2010) 49 susceptibility loci (Anderson CA et al. Nat Genet 2011)

6. Page 6 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Epigenetic Concept Integrated control of transcription independently from the genetic code Mitotically heritable Mitotic inheritance: DNA methylation – DNMT1; histone modifications - ?; ncRNAs - ? http://www.epigenetics.ch/mirna.html Autoregulatory proteins

7. Page 7 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics DNA Methylation, a Nutritionally Responsive Epigenetic Process Gene silencing Mammalian one carbon poolNutrition/environment -p-C-p-G-p- (CpG di-nucleotide)

8. Page 8 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Nutrition, DNA Methylation, and Phenotype Waterland RA et al. MCB 2003 A vy /a Methyl-donor diet Metastable epiallele Stochasticity

9. Page 9 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics

10. Page 10 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Working model Transient nutritional/environmental exposures Critical periods of development Permanent changes in the biological components of IBD Nutritionally/environmentally induced predisposition to IBD (if early development – detectable in all tissues – metastable epiallele for IBDs)

11. Page 11 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics PBL DNA methylation association in IBDs? 2 approaches ‐ MSAM on MZ twins concordant (CD: 4; UC: 3) and discordant (CD: 4; UC: 7) for IBD Validation (bisulfite pyrosequencing in independent cohort of adult patients) ‐ Illumina 450K Infinium Methylation BeadChips: 48 samples: discordant MZ twins (CD:3; UC:3) and treatment naive pediatric cases of IBD (CD:14; UC:8), controls (n=14) Validation in expanded cohort

12. Page 12 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics One single association in PBMCs TEPP :testis, prostate and placenta-expressed protein

13. Page 13 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics PBL DNA methylation association in IBD? Microarray interrogation of IBD dependent DNA methylation from PBLs is less likely to yield significant results More detailed and/or selective approaches may be useful

14. Page 14 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Selective Approach from PBL DNA Inflamm Bowel Dis. 2011 Oct 21. [Epub ahead of print]

15. Page 15 xxx00.#####.ppt 8/23/2015 4:50:59 PM Pediatrics Acknowledgements Dorottya Nagy-Szakal Ronald A. Harris Sabina Mir Harry Siegele, Sahna Reddy Natalia Pedersen,Jiri Bronsky, Pia Munkholm, Cathrine Jespersgaard, Paal Skytt Andersen, Bela Melegh, Tine Jess C. Wayne Smith George Ferry Mark A. Gilger Antone Opekun Funding: Broad Foundation, Baylor CHRC