Disrupting the TRIB3-SQSTM1 interaction reduces liver fibrosis by restoring autophagy and suppressing exosome-mediated HSC activation
Disrupting the TRIB3-SQSTM1 interaction reduces liver fibrosis by restoring autophagy and suppressing exosome-mediated HSC activation
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破坏 TRIB3-SQSTM1 相互作用可通过恢复自噬和抑制外泌体介导的 HSC 激活来减少肝纤维化
DOI：
10.1080/15548627.2019.1635383
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发表时间：
2020
期刊：
影响因子：
13.3
通讯作者：
Hu Zhuo-Wei
中科院分区：
文献类型：
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作者：
Zhang Xiao-Wei;Zhou Ji-Chao;Peng Dian;Hua Fang;Li Ke;Yu Jiao-Jiao;Lv Xiao-Xi;Cui Bing;Liu Shan-Shan;Yu Jin-Mei;Wang Feng;Jin Cai-Cai;Yang Zhao-Na;Zhao Chen-Xi;Hou Xue-Ying;Huang Bo;Hu Zhuo-Wei
自噬/巨噬功能受损参与了肝纤维化的发病机制。然而，异常的自噬如何促进纤维化还远未了解。在这里，我们的目的是确定一个以前未揭示的促纤维化机制的应激蛋白TRIB 3（tribbles pseudokinase 3）介导的自噬功能障碍。人纤维化肝组织获自经历开放性手术修复过程的肝硬化患者。在胆管结扎（BDL）或硫代乙酰胺（TAA）注射诱导的肝纤维化小鼠模型中评估TRIB 3的功能意义。人肝纤维化组织中TRIB 3和选择性自噬受体SQSTM 1/p62（隔离体1）的表达水平高于非纤维化组织，并且TRIB 3和SQSTM 1的表达升高在纤维化组织中呈正相关。沉默Trib 3保护免受实验诱导的肝纤维化，伴随着纤维化肝组织中恢复的自噬活性。TRIB 3与SQSTM 1相互作用，阻碍了SQSTM 1与MAP 1 LC 3/LC 3的结合，导致SQSTM 1聚集体的聚集，阻碍了自噬通量。TRIB 3介导的自噬损伤不仅抑制了晚期内体的自噬降解，而且还促进了肝细胞分泌富含INHBA/激活素A的外泌体，从而导致肝星状细胞（HSC）（肝纤维化的效应细胞）的迁移、增殖和激活。破坏TRIB 3-SQSTM 1与特定螺旋肽的相互作用，通过恢复肝细胞和HSC中的自噬通量，对肝纤维化产生有效的保护作用。总之，应激升高的TRIB 3表达通过与SQSTM 1相互作用并干扰其在肝实质细胞中的功能并激活HSC来促进肝纤维化。靶向这种相互作用是治疗纤维增生性肝病的一种有前途的策略。缩略语：3-甲基丙烯酸：3-甲基腺嘌呤; AV：腺相关病毒; ACTA 2/α-SMA：肌动蛋白，α 2，平滑肌，主动脉; BDL：胆管结扎; BECN 1/Beclin 1：Beclin 1，自噬相关; CHX：放线菌酮; CQ：氯喹; Edu：5-乙炔基-2-脱氧尿苷; ESCRT：转运所需的内体分选复合物; HSC：肝星状细胞; ILV：腔内囊泡; LAMP 1：溶酶体相关膜蛋白1; MAP 1 LC 3/LC 3：微管相关蛋白1轻链3; MVB：多泡体; PIK 3C 3：磷脂酰肌醇3-激酶，催化亚基3型; PPI：蛋白质-蛋白质相互作用; SQSTM 1/p62：多价螯合体1; TAA：硫代乙酰胺; TEM：透射电子显微镜; TGFB 1/TGFβ1：转化生长因子β 1; TLR 2：Toll样受体2; TRIB3：tribbles假激酶3
ABSTRACT Impaired macroautophagy/autophagy is involved in the pathogenesis of hepatic fibrosis. However, how aberrant autophagy promotes fibrosis is far from understood. Here, we aimed to define a previously unrevealed pro-fibrotic mechanism for the stress protein TRIB3 (tribbles pseudokinase 3)-mediated autophagy dysfunction. Human fibrotic liver tissues were obtained from patients with cirrhosis who underwent an open surgical repair process. The functional implications of TRIB3 were evaluated in mouse models of hepatic fibrosis induced by bile duct ligation (BDL) or thioacetamide (TAA) injection. Human fibrotic liver tissues expressed higher levels of TRIB3 and selective autophagic receptor SQSTM1/p62 (sequestosome 1) than nonfibrotic tissues and the elevated expression of TRIB3 and SQSTM1 was positively correlated in the fibrotic tissues. Silencing Trib3 protected against experimentally induced hepatic fibrosis, accompanied by restored autophagy activity in fibrotic liver tissues. Furthermore, TRIB3 interacted with SQSTM1 and hindered its binding to MAP1LC3/LC3, which caused the accumulation of SQSTM1 aggregates and obstructed autophagic flux. The TRIB3-mediated autophagy impairment not only suppressed autophagic degradation of late endosomes but also promoted hepatocellular secretion of INHBA/Activin A-enriched exosomes which caused migration, proliferation and activation of hepatic stellate cells (HSCs), the effector cells of liver fibrosis. Disrupting the TRIB3-SQSTM1 interaction with a specific helical peptide exerted potent protective effects against hepatic fibrosis by restoring autophagic flux in hepatocytes and HSCs. Together, stress-elevated TRIB3 expression promotes hepatic fibrosis by interacting with SQSTM1 and interfering with its functions in liver-parenchymal cells and activating HSCs. Targeting this interaction is a promising strategy for treating fibroproliferative liver diseases. Abbreviations: 3-MA: 3-methyladenine; AAV: adeno-associated virus; ACTA2/α-SMA: actin, alpha 2, smooth muscle, aorta; BDL: bile duct ligation; BECN1/Beclin 1: beclin 1, autophagy related; CHX: cycloheximide; CQ: chloroquine; Edu: 5-ethynyl-2-deoxyuridine; ESCRT: endosomal sorting complexes required for transport; HSC: hepatic stellate cell; ILV: intralumenal vesicle; LAMP1: lysosomal-associated membrane protein 1; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MVB: multivesicular body; PIK3C3: phosphatidylinositol 3-kinase, catalytic subunit type 3; PPI: protein-protein interaction; SQSTM1/p62: sequestosome 1; TAA: thioacetamide; TEM: transmission electron microscopy; TGFB1/TGFβ1: transforming growth factor, beta 1; TLR2: toll-like receptor 2; TRIB3: tribbles pseudokinase 3