Senescent cholangiocytes release extracellular vesicles that alter target cell phenotype via the epidermal growth factor receptor

作者：Mohammed S. Al Suraih, Christy E. Trussoni, Patrick L. Splinter, Nicholas F. LaRusso, Steven P. O’Hara · 发表于：Liver International · 年份：2020 · DOI：10.1111/liv.14569 · 被引用次数：40 · 研究领域：Extracellular vesicles in disease、Liver Diseases and Immunity、MicroRNA in disease regulation

Abstract Background & Aims Primary sclerosing cholangitis (PSC) is a chronic liver disease characterized by peribiliary inflammation and fibrosis. Cholangiocyte senescence is a prominent feature of PSC. Here, we hypothesize that extracellular vesicles (EVs) from senescent cholangiocytes influence the phenotype of target cells. Methods EVs were isolated from normal human cholangiocytes (NHCs), cholangiocytes from PSC patients and NHCs experimentally induced to senescence. NHCs, malignant human cholangiocytes (MHCs) and monocytes were exposed to 10 8 EVs from each donor cell population and assessed for proliferation, MAPK activation and migration. Additionally, we isolated EVs from plasma of wild‐type and Mdr2 −/− mice (a murine model of PSC), and assessed mouse monocyte activation. Results EVs exhibited the size and protein markers of exosomes. The number of EVs released from senescent human cholangiocytes was increased; similarly, the EVs in plasma from Mdr2 −/− mice were increased. Additionally, EVs from senescent cholangiocytes were enriched in multiple growth factors, including EGF. NHCs exposed to EVs from senescent cholangiocytes showed increased NRAS and ERK1/2 activation. Moreover, EVs from senescent cholangiocytes promoted proliferation of NHCs and MHCs, findings that were blocked by erlotinib, an EGF receptor inhibitor. Furthermore, EVs from senescent cholangiocytes induced EGF‐dependent Interleukin 1‐beta and Tumour necrosis factor expression and migration of hu...