Deletion of Tgm2 suppresses BMP-mediated hepatocyte-to-cholangiocyte metaplasia in ductular reaction

Abstract

Transglutaminase 2 (Tgm2) plays an essential role in hepatic repair following prolonged toxic injury. During cholestatic liver injury, the intrahepatic cholangiocytes undergo dynamic tissue expansion and remodelling, referred to as ductular reaction (DR), which is crucial for liver regeneration. However, the molecular mechanisms governing the dynamics of active cells in DR are still largely unclear. Here, we generated Tgm2-knockout mice (Tgm2^−/−) and Tgm2-^CreERT2-Rosa26-mTmG flox/flox (Tgm2^CreERT2-R26T/G^f/f) mice and performed a three-dimensional (3D) collagen gel culture of mouse hepatocytes to demonstrate how Tgm2 signalling is involved in DR to remodel intrahepatic cholangiocytes. Our results showed that the deletion of Tgm2 adversely affected the functionality and maturity of the proliferative cholangiocytes in DR, thus leading to more severe cholestasis during DDC-induced liver injury. Additionally, Tgm2 hepatocytes played a crucial role in the regulation of DR through metaplasia. We unveiled that Tgm2 regulated H3K4me3Q5ser via serotonin to promote BMP signalling activation to participate in DR. Besides, we revealed that the activation or inhibition of BMP signalling could promote or suppress the development and maturation of cholangiocytes in DDC-induced DR. Furthermore, our 3D collagen gel culture assay indicated that Tgm2 was vital for the development of cholangiocytes in vitro. Our results uncovered a considerable role of BMP signalling in controlling metaplasia of Tgm2 hepatocytes in DR and revealed the phenotypic plasticity of mature hepatocytes.

References

[1] KoppJL, GrompeM, SanderM. Stem cells versus plasticity in liver and pancreas regeneration. Nat Cell Biol. 2016;18:238-245.

[2] MichalopoulosGK. Liver regeneration: alternative epithelial pathways. Int J Biochem Cell Biol. 2011;43:173-179.

[3] SatoK, Marzioni M, MengF, FrancisH, GlaserS, AlpiniG. Ductular reaction in liver diseases: pathological mechanisms and translational significances. Hepatology. 2019;69:420-430.

[4] KoS, Russell JO, MolinaLM, MongaSP. Liver progenitors and adult cell plasticity in hepatic injury and repair: knowns and unknowns. Annu Rev Pathol. 2020;15:23-50.

[5] OkadaH, YamadaM, KamimotoK, et al. The transcription factor Klf5 is essential for intrahepatic biliary epithelial tissue remodeling after cholestatic liver injury. J Biol Chem. 2018;293:6214-6229.

[6] IshikawaT, FactorVM, MarquardtJU, et al. Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice. Hepatology. 2012;55:1215-1226.

[7] LuWY, BirdTG, BoulterL, et al. Hepatic progenitor cells of biliary origin with liver repopulation capacity. Nat Cell Biol. 2015;17:971-983.

[8] TakaseHM, ItohT, InoS, et al. FGF7 is a functional niche signal required for stimulation of adult liver progenitor cells that support liver regeneration. Genes Dev. 2013;27:169-181.

[9] JakubowskiA, Ambrose C, ParrM, et al. TWEAK induces liver progenitor cell proliferation. J Clin Invest. 2005;115:2330-2340.

[10] MichalopoulosGK, KhanZ. Liver stem cells: experimental findings and implications for human liver disease. Gastroenterology. 2015;149:876-882.

[11] SchaubJR, Huppert KA, KurialSNT, et al. De novo formation of the biliary system by TGFβ-mediated hepatocyte transdifferentiation. Nature. 2018;557:247-251.

[12] TarlowBD, PelzC, NauglerWE, et al. Bipotential adult liver progenitors are derived from chronically injured mature hepatocytes. Cell Stem Cell. 2014;15:605-618.

[13] SekiyaS, SuzukiA. Hepatocytes, rather than cholangiocytes, can be the major source of primitive ductules in the chronically injured mouse liver. Am J Pathol. 2014;184:1468-1478.

[14] KamimotoK, KanekoK, KokCYY, Okada H, MiyajimaA, ItohT. Heterogeneity and stochastic growth regulation of biliary epithelial cells dictate dynamic epithelial tissue remodeling. Elife. 2016;5:e15034.

[15] TatsukawaH, SanoT, FukayaY, et al. Dual induction of caspase 3-and transglutaminase-dependent apoptosis by acyclic retinoid in hepatocellular carcinoma cells. Mol Cancer. 2011;10:4.

[16] TatsukawaH, FukayaY, FramptonG, et al. Role of transglutaminase 2 in liver injury via cross-linking and silencing of transcription factor Sp1. Gastroenterology. 2009;136:1783-95.e10.

[17] LaurenziVD, MelinoG. Gene disruption of tissue transglutaminase. Mol Cell Biol. 2001;21:148-155.

[18] StrnadP, HaradaM, SiegelM, et al. Transglutaminase 2 regulates mallory body inclusion formation and injury-associated liver enlargement. Gastroenterology. 2007;132:1515-1526.

[19] NardacciR, IaconoOL, CiccosantiF, et al. Transglutaminase type II plays a protective role in hepatic injury. Am J Pathol. 2003;162:1293-1303.

[20] FuriniG, Schroeder N, HuangL, et al. Proteomic profiling reveals the Transglutaminase-2 externalization pathway in kidneys after unilateral ureteric obstruction. J Am Soc Nephrol. 2018;29:880-905.

[21] BeuersU, Wolters F, ElferinkRPJO. Mechanisms of pruritus in cholestasis: understanding and treating the itch. Nat Rev Gastroenterol Hepatol. 2023;20:26-36.

[22] SchellingJR. Tissue transglutaminase inhibition as treatment for diabetic glomerular scarring: it’s good to be glueless. Kidney Int. 2009;76:363-365.

[23] JunkinsRD, McCormick C, LinTJ. The emerging potential of autophagy-based therapies in the treatment of cystic fibrosis lung infections. Autophagy. 2014;10:538-547.

[24] ElßnerC, Goeppert B, LongerichT, et al. Nuclear translocation of RELB is increased in diseased human liver and promotes Ductular reaction and biliary fibrosis in mice. Gastroenterology. 2019;156:1190-1205.e14.

[25] BoyerJL, SorokaCJ. Bile formation and secretion: an update. J Hepatol. 2021;75:190-201.

[26] IsseK, Lesniak A, GramaK, et al. Preexisting epithelial diversity in normal human livers: a tissue-tethered cytometric analysis in portal/periportal epithelial cells. Hepatology. 2013;57:1632-1643.

[27] GouwASH, Clouston AD, TheiseND. Ductular reactions in human liver: diversity at the interface. Hepatology. 2011;54:1853-1863.

[28] FuJ, YangQ, SaiK, et al. TGM2 inhibition attenuates ID1 expression in CD44-high glioma-initiating cells. Neuro Oncol. 2013;15:1353-1365.

[29] KyritsiK, ChenL, O’BrienA, et al. Modulation of the tryptophan hydroxylase 1/monoamine oxidase-a/5-Hydroxytryptamine/5-Hydroxytryptamine receptor 2A/2B/2C Axis regulates biliary proliferation and liver fibrosis during cholestasis. Hepatology. 2020;71:990-1008.

[30] OmenettiA, YangL, GainetdinovRR, et al. Paracrine modulation of cholangiocyte serotonin synthesis orchestrates biliary remodeling in adults. Am J Physiol Gastrointest Liver Physiol. 2011;300:G303-G315.

[31] CervantesM, Sassone-Corsi P. Modification of histone proteins by serotonin in the nucleus. Nature. 2019;567:464-465.

[32] FarrellyLA, Thompson RE, ZhaoS, et al. Histone serotonylation is a permissive modification that enhances TFIID binding to H3K4me3. Nature. 2019;567:535-539.

[33] RossiJM, DunnNR, HoganBL, Zaret KS. Distinct mesodermal signals, including BMPs from the septum transversum mesenchyme, are required in combination for hepatogenesis from the endoderm. Genes Dev. 2001;15:1998-2009.

[34] ChoiTY, KhaliqM, TsurusakiS, et al. Bmp signaling governs biliary-driven liver regeneration in zebrafish via Tbx2b and Id2a. Hepatology. 2017;66:1616-1630.

[35] SugimotoH, YangC, LeBleuVS, et al. BMP-7 functions as a novel hormone to facilitate liver regeneration. FASEB J. 2007;21:256-264.

[36] PetersenMA, RyuJK, ChangKJ, et al. Fibrinogen activates BMP signaling in oligodendrocyte progenitor cells and inhibits remyelination after vascular damage. Neuron. 2017;96:1003-1012.e7.

[37] NagahamaY, SoneM, ChenX, et al. Contributions of hepatocytes and bile Ductular cells in Ductular reactions and remodeling of the biliary system after chronic liver injury. Am J Pathol. 2014;184:3001-3012.

[38] NishikawaY, DoiY, WatanabeH, et al. Transdifferentiation of mature rat hepatocytes into bile duct-like cells in vitro. Am J Pathol. 2005;166:1077-1088.

[39] SoneM, Nishikawa Y, NagahamaY, et al. Recovery of mature hepatocytic phenotype following bile ductular transdifferentiation of rat hepatocytes in vitro. Am J Pathol. 2012;181:2094-2104.

[40] NishikawaY, SoneM, NagahamaY, et al. Tumor necrosis factor-α promotes bile ductular transdifferentiation of mature rat hepatocytes in vitro. J Cell Biochem. 2013;114:831-843.

[41] LaiTS, LiuY, TuckerT, et al. Identification of chemical inhibitors to human tissue transglutaminase by screening existing drug libraries. Chem Biol. 2008;15:969-978.

[42] OkabeH, YangJ, SylakowskiK, et al. Wnt signaling regulates hepatobiliary repair following cholestatic liver injury in mice. Hepatology. 2016;64:1652-1666.

[43] KamimotoK, NakanoY, KanekoK, Miyajima A, ItohT. Multidimensional imaging of liver injury repair in mice reveals fundamental role of the ductular reaction. Commun Biol. 2020;3:289.

[44] KimKH, ChenCC, AlpiniG, Lau LF. CCN1 induces hepatic ductular reaction through integrin αvβ鈧?mediated activation of NF-κB. J Clin Invest. 2015;125:1886-1900.

[45] BaiocchiL, LeSageG, GlaserS, Alpini G. Regulation of cholangiocyte bile secretion. J Hepatol. 1999;31:179-191.

[46] GongAY, TietzPS, MuffMA, et al. Somatostatin stimulates ductal bile absorption and inhibits ductal bile secretion in mice via SSTR2 on cholangiocytes. Am J Physiol Cell Physiol. 2003;284:C1205-C1214.

[47] ReichM, SpomerL, KlindtC, et al. Downregulation of TGR5 (GPBAR1) in biliary epithelial cells contributes to the pathogenesis of sclerosing cholangitis. J Hepatol. 2021;75:634-646.

[48] ZlotorynskiE. Histone serotonylation boosts neuronal transcription. Nat Rev Mol Cell Biol. 2019;20:323.

RIGHTS & PERMISSIONS

2024 The Authors. Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.