Processing of miR17-92 Cluster in Hepatic Stellate Cells Promotes Hepatic Fibrogenesis During Alcohol-Induced Injury.
Exposure to alcohol and its metabolites can initiate hepatic injury and fibrogenesis. Fibrosis is mediated through hepatic stellate cell (HSC) activation, leading to global changes in mRNA and microRNA (miR) expression. miRs are expressed in cells or shuttled to exosomes which can be detected in tissue culture media (TCM) and biological fluids. The mechanisms and function underlying the differential expression and processing of miRs and their downstream effects during hepatic injury remain poorly understood.



Expression of primary (pri)-miR17-92. and individual members of this cluster, miR17a, 18a, 19a, 20a, 19b, and 92, were examined in primary HSCs and human LX2 cells exposed to alcohol-conditioned media (CM), liver tissue from a rodent model of alcoholic injury, and in exosomes from TCM and plasma of rodent models and patients with alcoholic liver disease (ALD). miR expression was examined in HSCs transduced with an AAV2 vector carrying GFP-miR19b or GFP-control transgene under the collagen promoter.



Profibrotic markers were enhanced in primary HSCs and LX2 cells exposed to alcohol-CM, concomitant with decreased miR19b expression and a significant increase in pri-miR17-92. Increased pri-miR17-92 was confirmed in a rodent model of alcohol-induced liver injury. Individual members of the cluster were inversely proportionate in cells and exosomes. AAV2-mediated miR19b overexpression inhibited miR17-92 and altered expression of individual cluster members in cells and exosomes. Expression of individual miR17-92 cluster members in plasma exosomes isolated from patients with ALD was similar to that seen in a rodent model of alcoholic injury and in vitro.



Reintroduction of miR19b inhibits HSC activation and modulates expression of pri-miR17-92 and the inverse expression of individual cluster members in cells and exosomes. Better understanding of miR17-92 processing may provide mechanistic insights into the role of individual miRs and exosomes during hepatic injury, revealing new therapeutic targets.
Liver Pathobiology Laboratory, Department of Internal Medicine, Carolinas Medical Center, Charlotte, North Carolina.
Elizabeth
E
Brandon-Warner
Liver Pathobiology Laboratory, Department of Internal Medicine, Carolinas Medical Center, Charlotte, North Carolina.
Nicole A
NA
Feilen
Liver Pathobiology Laboratory, Department of Internal Medicine, Carolinas Medical Center, Charlotte, North Carolina.
Catherine R
CR
Culberson
Liver Pathobiology Laboratory, Department of Internal Medicine, Carolinas Medical Center, Charlotte, North Carolina.
Conroy O
CO
Field
Center for Liver Diseases and Liver Transplant, Carolinas Medical Center, Charlotte, North Carolina.
Andrew S
AS
deLemos
Center for Liver Diseases and Liver Transplant, Carolinas Medical Center, Charlotte, North Carolina.
Mark W
MW
Russo
Liver Pathobiology Laboratory, Department of Internal Medicine, Carolinas Medical Center, Charlotte, North Carolina.
Laura W
LW
Schrum
D000818:Animals; D000431:Ethanol / Q000494:pharmacology*; D055354:Exosomes / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D055166:Hepatic Stellate Cells / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008108:Liver Diseases, Alcoholic / Q000097:blood / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000096:biosynthesis / Q000097:blood / Q000187:drug effects*; D061251:Primary Cell Culture; D051381:Rats
D035683:MicroRNAs; D000431:Ethanol
Alcohol; Exosomes; Fibrosis; Hepatic Stellate Cells; Liver; MicroRNA; miR19b