Traf2 and NCK Interacting Kinase Is a Critical Regulator of Procollagen I Trafficking and Hepatic Fibrogenesis in Mice

Abstract

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-induced HSC apoptosis and thus is an attractive antifibrotic strategy. We designed an immunofluorescence-based 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-interacting kinase (TNIK) was identified and interrogated in vitro and in vivo using the TNIK kinase inhibitor NCB-0846 or RNA interference-mediated 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-0846 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 in vivo. Mice receiving NCB-0846 displayed reduced CCl 4 -induced fibrogenesis compared to CCl 4 alone, although α-smooth muscle actin levels were unaltered. Conclusions: Our siRNA screen effectively identified TNIK as a key kinase involved in procollagen I trafficking in vitro and hepatic fibrogenesis in vivo. (Hepatology Communications 2022;6:593-609).

TNIK regulation of interferon signaling and endothelial cell response to virus infection

“…Collagen I, which plays a vital role in the formation of bone, skin, and tendon formation, relies on efficient secretion facilitated by the ER-to-Golgi transport of procollagen I. Research has demonstrated that the deletion or depletion of TNIK disrupts the secretion of procollagen I without affecting intracellular procollagen I protein levels ( 48 ). This disruption suggests that TNIK may be involved in viral entry and replication, particularly since some viruses exploit host secretory pathways, including the ER-to-Golgi transport.…”

TNIK drives castration-resistant prostate cancer via phosphorylating EGFR

“…Interestingly, NCB-0846 can disrupt the transport and secretion of type I procollagen and inhibit the production of matrix proteins to regulate liver fibrosis. 39 In PCa, matrix proteins can induce tumor cell metastasis and adhesion to bone cells. 39 Although previous studies have demonstrated that NCB-0846 has a good therapeutic effect in ERG-positive PCa, whether it has the effect of regulating PCa EMT and bone metastasis has not been verified.…”

Identification of Key Genes and Underlying Mechanisms in Cystic Fibrosis and its Associated Complications Based on Bioinformatics Analysis of Next Generation Sequencing Data Analysis

“…Accumulating evidence shows that MMP9 [133], S100A12 [148], HP (haptoglobin) [149], OSM (oncostatin M) [150], PRDM5 [151], TSPO (translocator protein) [152], IL18RAP [153], ADAMDEC1 [154], IL1RN [155], SCN9A [156], FADS2 [157], NCF4 [158], SERPINB1 [159], TNFRSF13B [160], IL2RB [161], DLG5 [162], FASLG (Fas ligand) [163], GPR68 [164], IL2RA [161], TCF4 [165], TAGAP (T cell activation RhoGTPase activating protein) [166], ABCB1 [167], FCRL3 [168], ITGB7 [169], PTGER4 [170] and TRAF3IP2 [171] are altered expression in gastrointestinal complications. Recent studies reported that MMP9 [133], S100A12 [172], IGF2 [173], GPR84 [174], SOCS3 [175], IGFBP2 [176], HP (haptoglobin) [177], MMP8 [178], OSM (oncostatin M) [179], TLR5 [180], S100A9 [181], PLIN5 [182], NRG1 [183], LCN2 [184], TSPO (translocator protein) [185], PLAU (plasminogen activator, urokinase) [186], PPBP (pro-platelet basic protein) [187], UPP1 [188], ALOX5 [189], IL1RN [190], CYP1B1 [191], DGAT2 [192], MSRA (methionine sulfoxidereductase A) [193], ADAM9 [194], CPEB4 [195], IRS2 [196], FADS2 [197], SLC22A4 [198], PFKFB3 [199], CTSD (cathepsin D) [200], ADAM12 [201], CD80 [202], ERBB2 [203], FASLG (Fas ligand) [204], CNR2 [205], SOCS2 [206], ABCB1 [207], CD74 [208], FCRL3 [209], PARP1 [210], TXNIP (thioredoxin interacting protein) [211], TULP3 [212], HSPA8 [213], TNIK (TRAF2 and NCK interacting kinase) [214], PDCD4 [215…”