{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T12:47:27Z","timestamp":1768567647890,"version":"3.49.0"},"posted":{"date-parts":[[2024,9,28]]},"group-title":"Gastroenterology","reference-count":41,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,9,28]],"date-time":"2024-09-28T00:00:00Z","timestamp":1727481600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,9,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background and Aims<\/jats:title>\n                  <jats:p>\n                    A cohort of patients with inflammatory bowel disease (IBD) exhibit expansion of the gut pathobiont, adherent-invasive\n                    <jats:italic>E. coli<\/jats:italic>\n                    (AIEC). Loss of activity of the IBD susceptibility gene, protein tyrosine phosphatase type 2 (\n                    <jats:italic>PTPN2<\/jats:italic>\n                    ), results in dysbiosis of the gut microbiota both in human subjects and mice. Further, constitutive\n                    <jats:italic>Ptpn2<\/jats:italic>\n                    knock-out (\n                    <jats:italic>Ptpn2<\/jats:italic>\n                    -KO) mice display expansion of AIEC compared to wildtype littermates. CEACAM6, a host cell surface glycoprotein, is exploited by AIEC to attach to and enter intestinal epithelial cells (IECs). Here, we investigate the role of IEC-specific\n                    <jats:italic>PTPN2<\/jats:italic>\n                    in restricting AIEC invasion.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    Biopsies from IBD patients heterozygous (CT) or homozygous (CC) for the\n                    <jats:italic>PTPN2<\/jats:italic>\n                    SNP (single nucleotide polymorphism)\n                    <jats:italic>rs1893217<\/jats:italic>\n                    were processed for immunohistochemistry. HT-29 intestinal epithelial cells (IEC) were transfected with control shRNA (\n                    <jats:italic>PTPN2<\/jats:italic>\n                    -CTL), or a shRNA targeted towards\n                    <jats:italic>PTPN2<\/jats:italic>\n                    (\n                    <jats:italic>PTPN2<\/jats:italic>\n                    -KD). The\n                    <jats:italic>rs1893217<\/jats:italic>\n                    SNP was inserted (\n                    <jats:italic>PTPN2<\/jats:italic>\n                    -KI), or a complete knock-out of\n                    <jats:italic>PTPN2<\/jats:italic>\n                    (\n                    <jats:italic>PTPN2<\/jats:italic>\n                    - KO) was generated, with CRISPR-Cas9 gene editing of Caco-2BBe IEC lines. Adherence and invasion assays were performed with either the human IBD AIEC isolate, LF82, or a novel fluorescent-tagged mouse adherent-invasive\n                    <jats:italic>E. coli<\/jats:italic>\n                    (\n                    <jats:italic>m<\/jats:italic>\n                    AIEC\n                    <jats:sup>red<\/jats:sup>\n                    ) at multiplicity of infection (MOI) of 10. IL-6 and the pan-JAK inhibitor tofacitinib were administered to interrogate JAK-STAT signaling. Protein expression was determined by western blotting and densitometry.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    CEACAM6 expression was elevated (colon and ileum) in IBD patients carrying the\n                    <jats:italic>PTPN2 rs1893217<\/jats:italic>\n                    SNP (CT, CC) compared to wildtype (TT) IBD patients. HT-29 and Caco-2BBe cell lines deficient in\n                    <jats:italic>PTPN2<\/jats:italic>\n                    expressed significantly higher levels of CEACAM6. Further,\n                    <jats:italic>PTPN2<\/jats:italic>\n                    -KI and\n                    <jats:italic>PTPN2<\/jats:italic>\n                    -KO cell lines also displayed greater adherence and invasion by AIEC LF82 and higher\n                    <jats:italic>m<\/jats:italic>\n                    AIEC\n                    <jats:sup>red<\/jats:sup>\n                    invasion. CEACAM6 expression was further elevated after administration of IL-6 in\n                    <jats:italic>PTPN2<\/jats:italic>\n                    \u2013deficient cell lines compared to untreated controls. Silencing of STAT1 and 3 partially reduced CEACAM6 protein expression. Tofacitinib significantly reduced the elevated CEACAM6 protein expression and the higher AIEC adherence and invasion in\n                    <jats:italic>PTPN2<\/jats:italic>\n                    -KI and PTPN2-KO cell lines compared to DMSO controls.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                    Our findings highlight a crucial role for\n                    <jats:italic>PTPN2<\/jats:italic>\n                    in restricting pathobiont entry into host cells. Our study also describes a role for the FDA-approved drug, tofacitinib (Xeljanz) in correcting the JAK-STAT- mediated over-expression of CEACAM6, used by pathobionts as an entry portal into host cells. These findings suggest a role for JAK-inhibitors in mitigating AIEC colonization in IBD-susceptible hosts.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.09.26.24314341","type":"posted-content","created":{"date-parts":[[2024,9,28]],"date-time":"2024-09-28T16:15:14Z","timestamp":1727540114000},"source":"Crossref","is-referenced-by-count":1,"title":["The JAK inhibitor, Tofacitinib, Corrects the Overexpression of CEACAM6 and Limits Susceptibility to AIEC Caused by Reduced Activity of the IBD Associated Gene,\n                  <i>PTPN2<\/i>"],"prefix":"10.64898","author":[{"given":"Pritha","family":"Chatterjee","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vinicius","family":"Canale","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephanie J.","family":"King","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ali","family":"Shawki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hillmin","family":"Lei","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Haddad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Casey M.","family":"Gries","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dermot P.B.","family":"McGovern","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James","family":"Borneman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Declan F.","family":"McCole","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"issue":"1","key":"2024100306250575000_2024.09.26.24314341v1.1","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/S2468-1253(19)30358-9","article-title":"Global burden of inflammatory bowel disease","volume":"5","year":"2020","journal-title":"The Lancet Gastroenterology & Hepatology"},{"key":"2024100306250575000_2024.09.26.24314341v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2025-2"},{"issue":"3","key":"2024100306250575000_2024.09.26.24314341v1.3","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/S2468-1253(16)30111-X","article-title":"Genetics of inflammatory bowel disease: beyond NOD2","volume":"2","year":"2017","journal-title":"The lancet Gastroenterology & hepatology"},{"key":"2024100306250575000_2024.09.26.24314341v1.4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.20.100301.064816"},{"key":"2024100306250575000_2024.09.26.24314341v1.5","doi-asserted-by":"publisher","DOI":"10.3109\/00365528909091339"},{"key":"2024100306250575000_2024.09.26.24314341v1.6","doi-asserted-by":"publisher","DOI":"10.1159\/000228553"},{"issue":"43","key":"2024100306250575000_2024.09.26.24314341v1.7","doi-asserted-by":"crossref","first-page":"12296","DOI":"10.3748\/wjg.v21.i43.12296","article-title":"Genetics of inflammatory bowel disease from multifactorial to monogenic forms","volume":"21","year":"2015","journal-title":"World journal of gastroenterology"},{"key":"2024100306250575000_2024.09.26.24314341v1.8","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3760."},{"key":"2024100306250575000_2024.09.26.24314341v1.9","doi-asserted-by":"publisher","DOI":"10.1038\/ng.381"},{"key":"2024100306250575000_2024.09.26.24314341v1.10","doi-asserted-by":"publisher","DOI":"10.1002\/ibd.21913"},{"key":"2024100306250575000_2024.09.26.24314341v1.11","doi-asserted-by":"crossref","unstructured":"MR, S., et al., The Clinical Relevance of the IBD-Associated Variation Within the Risk Gene Locus Encoding Protein Tyrosine Phosphatase Non-Receptor Type 2 in Patients of the Swiss IBD Cohort. 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