1. SHIP1 deficiency in IBD Sandra Fernandes 1, James C. Ryan 2, William G. Kerr 1 1 SUNY-Upstate Medical University, Syracuse, NY 2 UCSF/SFVA, San Francisco, CA Friday, December 13, 2013

2. DISCLOSURES Nothing to disclose

3. SHIP1 and PI3K/Akt pathway Fernandes & al., 2013

4. SHIP -/- mice Germline SHIP1-deficiency cause fatal eosinophilic pneumonia and Crohn’s-like ileitis in mice For more on SHIP1 and mucosal inflammation in mouse model, please visit poster P-188 Kerr & al., GUT, 2011

5. Study Design 25ml Blood CD and UC patients PBMC Protein Expression (WB) SHIP1 (2 Abs.) Actin, Tubulin miRNAmiSEQTotal RNAqRT-PCR SHIP1 (2 sets primers) RPLP1, PPIA DNA Sequencing exome 30 x700bp amplicons

6. Western Blotting SHIP1 : P1C1 + goat anti-mouse HRP, Actin : C-19 + donkey anti-goat HRP, Santa Cruz Biotechnologies

7. Reproducibility, Stability

8. Protein levels vs mRNA

10. Protein degradation or translation inhibition? Modified from Grabbe & al. 2011Ameres & Zamore, 2013

11. Take home message SHIP1-deficiency is found in 10/47 samples (>20% of samples) Both CD and UC SHIP1-deficiency is stable over months No correlation to treatments at time of sampling or to other clinical parameters No reduction in SHIP1 mRNA levels –Translational or post-translational

12. Acknowledgments William Kerr James Ryan and his team at UCSF/SFVA Neetu Srivastava Raki Sudan Bonnie Toms Robert Brooks Matt Gumbleton Sonia Iyer Sudha Neelam Christie Youngs Katie Miller Scott Stegemann Summer students CCFA (Senior Research Award to WGK) NIH Paige Arnold Butterfly Run Carol M. Baldwin Foundation