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Boyer, J. L.

Publications and source records attributed to Boyer, J. L..

2 recordsLinked to original sources

Human Vascularized Bile Duct-on-a Chip: A multi-cellular micro-physiological system for studying Primary Sclerosing Cholangitis

Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease in which the bile ducts of the liver become inflamed and scarred. Scarred bile ducts eventually narrow and obstruct and can cause additional liver pathology including liver failure, repeated infections, and tumors. The pathogenesis of PSC remains largely unknown, partly due to difficulty in obtaining cholangiocytes and partly due to a paucity of in vitro models that capture the various factors contributing to disease progression. Here we report the development of a human vascularized bile duct-on-a-chip that models blood vessels and bile ducts structurally and functionally in three dimensions and includes cholangiocytes derived from control and PSC patient tissue and bile. The flow of blood and bile was modeled by perfusion of cell-lined channels, and cholangiocytes and endothelial cells displayed differential responses to perfusion. Normal and PSC cholangiocytes polarized normally, formed mature tight junctions and displayed similar permeability, comparable to ex vivo measurements. The model with PSC cholangiocytes, however, became more inflammatory than the normal under the stimulation of IL-17A, which induced PBMC and differentiated Th17 cells in the vascular channel to transmigrate more through the endothelial layer of the vascular compartment. In sum, this human vascularized bile duct-on-a-chip recapitulated the vascular-biliary interface structurally and functionally and represents a novel multicellular platform to study inflammatory and fibrotic cholangiopathies such as PSC.

bioengineering↗

Hepatic NFAT signaling regulates the expression of inflammatory cytokines in cholestasis

The inflammatory response plays an important role in cholestatic liver injury where bile acid (BA) induction of proinflammatory cytokines in hepatocytes is an initial pathophysiologic event. However, the signaling pathways involving BA stimulation of cytokine production remain to be elucidated. In this report, we examined the functional role of the Nuclear Factor of Activated T-cells (NFAT) in BA-induction of inflammatory genes in hepatic cells and cholestatic livers. We found that NFAT isoform c1 and c3 were expressed in human and mouse hepatocytes. When treated with cholestatic levels of BA, both human and mouse hepatocytes but not cholangiocytes increased NFATc3 nuclear translocation, associated with elevated mRNA levels of IL-8, Cxcl2, and Cxcl10 in these cells. Blocking NFAT activation with pathway-specific inhibitors (i.e. cyclosporine A, FK-506, KN-62 and Inca-6) or knocking down Nfatc3, significantly repressed BA-induction of these cytokines in mouse hepatocytes, including Ccl2, Cxcl2, Cxcl10, Icam1 and Egr1. Nuclear expression of NFATc3/Nfatc3 protein was also increased in cholestatic livers after bile duct ligation or in Abcb4-/- mice and in patients with primary biliary cholangitis and primary sclerosing cholangitis in association with tissue elevations of Cxcl2 and IL-8. Gene reporter assays and ChIP-PCR demonstrated that the NFAT response element in its promoter played a key role in BA-induced human IL-8 expression. Together our findings indicate that NFAT plays an important role in BA stimulation of hepatic cytokines in cholestasis and is a mechanism that may provide novel targets to reduce cholestatic liver injury.

molecular biology↗