A Frameshift in CSF2RB Predominant Among Ashkenazi Jews Increases Risk for Crohn's Disease and Reduces Monocyte Signaling via GM-CSF

• Type of Material: Article
• Authors: Ling-Shiang Chuang, Icahn School of Medicine at Mount Sinai; Nicole Villaverde, Icahn School of Medicine at Mount Sinai; Ken Y. Hui, Yale University; Arthur Mortha, Icahn School of Medicine at Mount Sinai; Adeeb Rahman, Icahn School of Medicine at Mount Sinai; Adam P. Levine, University College, London; Talin Haritunians, Yale University; Sok Meng Evelyn Ng, Yale University; Wei Zhang, Yale University; Nai-Yun Hsu, Icahn School of Medicine at Mount Sinai; Jody-Ann Facey, Icahn School of Medicine at Mount Sinai; Tramy Luong, Icahn School of Medicine at Mount Sinai; Heriberto Fernandez-Hernandez, Icahn School of Medicine at Mount Sinai; Dalin Li, Cedars-Sinai Medical Center; Manuel Rivas, Broad Institute; Elena R. Schiff, University College, London; Alexander Gusev, Harvard University; L. Phillip Schumm, University of Chicago; Beatrice M. Bowen, Yale University; Yashoda Sharma, Yale University; Kaida Ning, Yale University; Romain Remark, Icahn School of Medicine at Mount Sinai; Sacha Gnjatic, Icahn School of Medicine at Mount Sinai; Peter Legnani, Icahn School of Medicine at Mount Sinai; James George, Icahn School of Medicine at Mount Sinai; Bruce E. Sands, Icahn School of Medicine at Mount Sinai; Joanne M. Stempak, Mount Sinai Hospital; Lisa W. Datta, Johns Hopkins University; Seth Lipka, University of South Florida; Seymour Katz, New York University; Adam S. Cheifetz, Beth Israel Deaconess Medical Center; Nir Barzilai, Albert Einstein College of Medicine; Nikolas Pontikos, University College, London; Clara Abraham, Yale University; Marla J. Dubinsky, Icahn School of Medicine at Mount Sinai; Stephan Targan, Cedars-Sinai Medical Center; Kent Taylor, Harbor-UCLA Medical Center; Jerome I. Rotter, Harbor-UCLA Medical Center; Ellen J. Scherl, Cornell University; Robert J. Desnick, Icahn School of Medicine at Mount Sinai; Maria T. Abreu, University of Miami; Hongyu Zhao, Yale University; Gil Atzmon, Albert Einstein College of Medicine; Itsik Pe'er, Columbia University; Subramaniam Kugathasan, Emory University; Hakon Hakonarson, Childrens Hospital of Philadelphia; Jacob L. McCauley, University of Miami; Todd Lencz, North Shore – Long Island Jewish Health System; Ariel Darvasi, Hebrew University of Jerusalem; Vincent Plagnol, University College, London; Mark S. Silverberg, Mount Sinai Hospital; Alexio M. Muise, University of Toronto; Steven R. Brant, Johns Hopkins University; Mark J. Daly, Massachusetts General Hospital; Anthony W. Segal, University College, London; Richard H. Duerr, University of Pittsburgh; Miriam Merad, Icahn School of Medicine at Mount Sinai; Dermot P.B. McGovern, Cedars-Sinai Medical Center; Inga Peter, Icahn School of Medicine at Mount Sinai; Judy H. Cho, Icahn School of Medicine at Mount Sinai
• Language: English
• Date: 2016-10-01
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2016 AGA Institute
• Final Published Version (URL): http://dx.doi.org/10.1053/j.gastro.2016.06.045
• Title of Journal or Parent Work: Gastroenterology
• ISSN: 0016-5085
• Volume: 151
• Issue: 4
• Start Page: 710
• End Page: +
• Grant/Funding Information: Project investigators are supported by The Helmsley Charitable Trust (D.P.B.M.), The European Union (D.P.B.M.), The Crohn’s and Colitis Foundation of America (CCFA) (D.P.B.M.), The Joshua L. and Lisa Z. Greer Chair in IBD Genetics (D.P.B.M.), and grants, DK062413, DK046763-19, AI067068, HS021747 (D.P.B.M.).; Researchers at UCL were funded by the Wellcome Trust, Charles Wolfson Charitable Trust, and the Irwin Joffe Memorial Fellowship. IBD Research at Cedars-Sinai is supported by USPHS grant PO1 (DK046763) and the Cedars-Sinai F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute Research Funds.; The NIH (DK092235), Inflammatory Bowel Disease Genetics Consortium (DK062429), Genetic Research Center at the Icahn School of Medicine (DK062422), New York Crohn’s Foundation, Consortium ancillary RO1 (DK099097), U01 (DK062431), Inflammatory Bowel Disease Genetic Research Chair, RO1 (DK062420), RO1 (CA141743), the Atran Foundation and the Sanford J. Grossman Charitable Trust financially supported this work.
• Keywords: IBD; RECEPTOR; INFLAMMATORY-BOWEL-DISEASE; Risk Factor; ACTIVATION; Inflammatory Bowel Disease; ASSOCIATION; Life Sciences & Biomedicine; Gastroenterology & Hepatology; SUSCEPTIBILITY; AUTOANTIBODIES; Ethnic Variation; EXPRESSION; PROLIFERATION; COLONY-STIMULATING FACTOR; Science & Technology; CYTOMETRY DATA
• Research Categories: Biology, Biostatistics; Biology, Bioinformatics

Abstract

Background & Aims: Crohn's disease (CD) has the highest prevalence in Ashkenazi Jewish populations. We sought to identify rare, CD-associated frameshift variants of high functional and statistical effects. Methods: We performed exome sequencing and array-based genotype analyses of 1477 Ashkenazi Jewish individuals with CD and 2614 Ashkenazi Jewish individuals without CD (controls). To validate our findings, we performed genotype analyses of an additional 1515 CD cases and 7052 controls for frameshift mutations in the colony-stimulating factor 2–receptor β common subunit gene (CSF2RB). Intestinal tissues and blood samples were collected from patients with CD; lamina propria leukocytes were isolated and expression of CSF2RB and granulocyte-macrophage colony–stimulating factor–responsive cells were defined by adenomatous polyposis coli (APC) time-of-flight mass cytometry (CyTOF analysis). Variants of CSF2RB were transfected into HEK293 cells and the expression and functions of gene products were compared. Results: In the discovery cohort, we associated CD with a frameshift mutation in CSF2RB (P = 8.52 × 10 -4 ); the finding was validated in the replication cohort (combined P = 3.42 × 10 -6 ). Incubation of intestinal lamina propria leukocytes with granulocyte-macrophage colony–stimulating factor resulted in high levels of phosphorylation of signal transducer and activator of transcription (STAT5) and lesser increases in phosphorylation of extracellular signal–regulated kinase and AK straining transforming (AKT). Cells co-transfected with full-length and mutant forms of CSF2RB had reduced pSTAT5 after stimulation with granulocyte-macrophage colony–stimulating factor, compared with cells transfected with control CSF2RB, indicating a dominant-negative effect of the mutant gene. Monocytes from patients with CD who were heterozygous for the frameshift mutation (6% of CD cases analyzed) had reduced responses to granulocyte-macrophage colony–stimulating factor and markedly decreased activity of aldehyde dehydrogenase; activity of this enzyme has been associated with immune tolerance. Conclusions: In a genetic analysis of Ashkenazi Jewish individuals, we associated CD with a frameshift mutation in CSF2RB. Intestinal monocytes from carriers of this mutation had reduced responses to granulocyte-macrophage colony–stimulating factor, providing an additional mechanism for alterations to the innate immune response in individuals with CD.