{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T18:31:30Z","timestamp":1788373890194,"version":"build-2803163510"},"reference-count":50,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"3","license":[{"start":{"date-parts":[[2021,10,22]],"date-time":"2021-10-22T00:00:00Z","timestamp":1634860800000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":["Hepatol Commun"],"published-print":{"date-parts":[[2022,3]]},"abstract":"<jats:p>Hepatic fibrosis is driven by deposition of matrix proteins following liver injury. Hepatic stellate cells (HSCs) drive fibrogenesis, producing matrix proteins, including procollagen I, which matures into collagen I following secretion. Disrupting intracellular procollagen processing and trafficking causes endoplasmic reticulum stress and stress\u2010induced HSC apoptosis and thus is an attractive antifibrotic strategy. We designed an immunofluorescence\u2010based small interfering RNA (siRNA) screen to identify procollagen I trafficking regulators, hypothesizing that these proteins could serve as antifibrotic targets. A targeted siRNA screen was performed using immunofluorescence to detect changes in intracellular procollagen I. Tumor necrosis factor receptor associated factor 2 and noncatalytic region of tyrosine kinase\u2010interacting kinase (TNIK) was identified and interrogated <jats:italic toggle=\"yes\">in vitro<\/jats:italic> and <jats:italic toggle=\"yes\">in vivo<\/jats:italic> using the TNIK kinase inhibitor NCB\u20100846 or RNA interference\u2010mediated knockdown. Our siRNA screen identified nine genes whose knockdown promoted procollagen I retention, including the serine\/threonine kinase TNIK. Genetic deletion or pharmacologic inhibition of TNIK through the small molecule inhibitor NCB\u20100846 disrupted procollagen I trafficking and secretion without impacting procollagen I expression. To investigate the role of TNIK in liver fibrogenesis, we analyzed human and murine livers, finding elevated TNIK expression in human cirrhotic livers and increased TNIK expression and kinase activity in both fibrotic mouse livers and activated primary human HSCs. Finally, we tested whether inhibition of TNIK kinase activity could limit fibrogenesis <jats:italic toggle=\"yes\">in vivo<\/jats:italic>. Mice receiving NCB\u20100846 displayed reduced CCl<jats:sub>4<\/jats:sub>\u2010induced fibrogenesis compared to CCl<jats:sub>4<\/jats:sub> alone, although \u03b1\u2010smooth muscle actin levels were unaltered. <jats:italic toggle=\"yes\">Conclusions:<\/jats:italic> Our siRNA screen effectively identified TNIK as a key kinase involved in procollagen I trafficking <jats:italic toggle=\"yes\">in vitro<\/jats:italic> and hepatic fibrogenesis <jats:italic toggle=\"yes\">in vivo<\/jats:italic>.<\/jats:p>","DOI":"10.1002\/hep4.1835","type":"journal-article","created":{"date-parts":[[2021,10,22]],"date-time":"2021-10-22T10:44:36Z","timestamp":1634899476000},"page":"593-609","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":19,"title":["Traf2 and NCK Interacting Kinase Is a Critical Regulator of Procollagen I Trafficking and Hepatic Fibrogenesis in Mice"],"prefix":"10.1097","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4645-8717","authenticated-orcid":false,"given":"Samuel C.","family":"Buchl","sequence":"first","affiliation":[{"name":"Division of Gastroenterology and Hepatology,Mayo Clinic,Rochester,MN,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zachary","family":"Hanquier","sequence":"additional","affiliation":[{"name":"Department of Medical and Molecular Genetics,Indiana University School of Medicine,Indianapolis,IN,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew J.","family":"Haak","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biomedical Engineering,Mayo Clinic,Rochester,MN,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yvonne M.","family":"Thomason","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology,Indiana University School of Medicine,Indianapolis,IN,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9812-7573","authenticated-orcid":false,"given":"Robert C.","family":"Huebert","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology,Mayo Clinic,Rochester,MN,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7620-573X","authenticated-orcid":false,"given":"Vijay H.","family":"Shah","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology,Mayo Clinic,Rochester,MN,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7798-2274","authenticated-orcid":false,"given":"Jessica L.","family":"Maiers","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology,Indiana University School of 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