SHIP1 deficiency in IBD

Sandra Fernandes1, James C. Ryan2, William G. Kerr1

1SUNY-Upstate Medical University, Syracuse, NY2UCSF/SFVA, San Francisco, CA

Friday, December 13, 2013

DISCLOSURES

• Nothing to disclose

SHIP1 and PI3K/Akt pathway

Fernandes & al., 2013

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

Study Design25ml Blood

CD and UC

patients

PBMC

Protein Expression

(WB)

SHIP1

(2 Abs.)Actin,

Tubulin

miRNA

miSEQ

Total RNA

qRT-PCR

SHIP1 (2 sets

primers)

RPLP1, PPIA

DNA

Sequencing exome

30 x700bp amplicons

Western Blotting

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

Reproducibility, Stability

Protein levels vs mRNA

Protein levels vs mRNA

Protein degradation or translation inhibition?

Modified from Grabbe & al. 2011 Ameres & Zamore, 2013

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

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