Cholangiocyte Epithelial to Mesenchymal Transition (EMT) is a potential molecular mechanism driving ischemic cholangiopathy in liver transplantation.
Cholangiocyte Epithelial to Mesenchymal Transition (EMT) is a potential molecular mechanism driving ischemic cholangiopathy in liver transplantation.
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DOI：
10.1371/journal.pone.0246978
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Mangino MJ
中科院分区：
文献类型：
--
作者：
Wickramaratne N;Li R;Tian T;Khoraki J;Kang HS;Chmielewski C;Maitland J;Liebrecht LK;Fyffe-Freil R;Lindell SL;Mangino MJ
循环性死亡后捐赠（DCD）扩大了肝移植的供体池。然而，DCD肝移植后缺血性胆管病（IC）的预后较差。IC的分子机制目前尚不清楚，但可能依赖于缺血诱导的胆道上皮向间充质样细胞的遗传重编程。本研究的主要目的是确定暴露于DCD条件下胆管细胞是否经历上皮到间充质转化（EMT），以及这是否导致IC的表型。人类胆管细胞培养暴露于温暖和冷缺血时期，以模拟DCD肝脏捐赠。通过细胞迁移、细胞形态和差异基因表达检测EMT。采用免疫组织化学方法评价DCD条件下恢复的同种大鼠肝脏移植后EMT的变化。暴露于DCD条件下的人胆管细胞表现出与EMT一致的迁移行为和基因表达模式。E-cadherin和CK-7的表达下降，而N-cadherin、vimentin、TGFβ和SNAIL的表达上升，在暴露后24小时开始，1-3周达到峰值。缺血1周后，胆管细胞形态由缺血前的立方体（上皮细胞）变为梭形（间质细胞）。转化生长因子β (TGFβ)受体拮抗剂Galunisertib （1 μM）预处理细胞可阻断这些变化。最后，肝等移植物在UW溶液中冷藏20小时，恢复时热缺血（30分钟），移植后1周血浆胆红素升高，肝组织显示早期胆管细胞EMT的免疫组化证据。我们的研究结果表明，EMT发生在人胆管细胞暴露于DCD条件后，这可能是由自分泌来源的TGFβ的上游信号引发的，导致间充质特异性形态和迁移变化。
Donation after circulatory death (DCD) has expanded the donor pool for liver transplantation. However, ischemic cholangiopathy (IC) after DCD liver transplantation causes inferior outcomes. The molecular mechanisms of IC are currently unknown but may depend on ischemia-induced genetic reprograming of the biliary epithelium to mesenchymal-like cells. The main objective of this study was to determine if cholangiocytes undergo epithelial to mesenchymal transition (EMT) after exposure to DCD conditions and if this causally contributes to the phenotype of IC. Human cholangiocyte cultures were exposed to periods of warm and cold ischemia to mimic DCD liver donation. EMT was tested by assays of cell migration, cell morphology, and differential gene expression. Transplantation of syngeneic rat livers recovered under DCD conditions were evaluated for EMT changes by immunohistochemistry. Human cholangiocytes exposed to DCD conditions displayed migratory behavior and gene expression patterns consistent with EMT. E-cadherin and CK-7 expressions fell while N-cadherin, vimentin, TGFβ, and SNAIL rose, starting 24 hours and peaking 1–3 weeks after exposure. Cholangiocyte morphology changed from cuboidal (epithelial) before to spindle shaped (mesenchymal) a week after ischemia. These changes were blocked by pretreating cells with the Transforming Growth Factor beta (TGFβ) receptor antagonist Galunisertib (1 μM). Finally, rats with liver isografts cold stored for 20 hours in UW solution and exposed to warm ischemia (30 minutes) at recovery had elevated plasma bilirubin 1 week after transplantation and the liver tissue showed immunohistochemical evidence of early cholangiocyte EMT. Our findings show EMT occurs after exposure of human cholangiocytes to DCD conditions, which may be initiated by upstream signaling from autocrine derived TGFβ to cause mesenchymal specific morphological and migratory changes.
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影响因子：
6.2
作者：
Goldberg，David S.;Karp，Seth J.;Abt，Peter L.
通讯作者：
Abt，Peter L.
影响因子：
3.8
作者：
Skaro AI;Jay CL;Baker TB;Wang E;Pasricha S;Lyuksemburg V;Martin JA;Feinglass JM;Preczewski LB;Abecassis MM
通讯作者：
Abecassis MM
缺氧，上皮间质转变和TET介导的表观遗传变化。
影响因子：
3.9
作者：
Kao SH;Wu KJ;Lee WH
通讯作者：
Lee WH
影响因子：
9
作者：
Foley DP;Fernandez LA;Leverson G;Anderson M;Mezrich J;Sollinger HW;D'Alessandro A
通讯作者：
D'Alessandro A
影响因子：
7.3
作者：
Harada, Kenichi;Sato, Yasunori;Nakanuma, Yasuni
通讯作者：
Nakanuma, Yasuni