Fibrogenic Signaling Is Suppressed in Hepatic Stellate Cells through Targeting of Connective Tissue Growth Factor (CCN2) by Cellular or Exosomal MicroRNA-199a-5p.
Pathways of liver fibrosis are controlled by connective tissue growth factor (CCN2). In this study, CCN2 was identified as a target of miR-199a-5p, which was principally expressed in quiescent mouse hepatic stellate cells (HSCs) and directly suppressed production of CCN2. Up-regulated CCN2 expression in fibrotic mouse livers or in activated primary mouse HSCs was associated with miR-199a-5p down-regulation. MiR-199a-5p in quiescent mouse HSCs inhibited the activity of a wild-type CCN2 3' untranslated region (3'-UTR) but not of a mutant CCN2 3'-UTR lacking the miR-199a-5p-binding site. In activated mouse HSCs, CCN2, α-smooth muscle actin, and collagen 1(α1) were suppressed by a miR-199a-5p mimic, whereas in quiescent mouse HSCs, the inhibited CCN2 3'-UTR activity was blocked by a miR-199a-5p antagomir. CCN2 3'-UTR activity in human HSCs was reduced by a miR-199a-5p mimic. MiR-199a-5p was present at higher levels in exosomes from quiescent versus activated HSCs. MiR-199a-5p-containing exosomes were shuttled from quiescent mouse HSCs to activated mouse HSCs in which CCN2 3'-UTR activity was then suppressed. Exosomes from quiescent HSCs caused miR-199a-5p-dependent inhibition of CCN2, α-smooth muscle actin, or collagen 1(α1) in activated HSCs in vitro and bound to activated HSCs in vivo. Thus, CCN2 suppression by miR-199a-5p accounts, in part, for low-level fibrogenic gene expression in quiescent HSCs and causes dampened gene expression in activated HSCs after horizontal transfer of miR-199a-5p in exosomes from quiescent HSCs.
Center for Clinical and Translational Research, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
Li
L
Chen
Center for Clinical and Translational Research, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
Ruju
R
Chen
Center for Vaccines and Immunity, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
Victoria M
VM
Velazquez
Center for Clinical and Translational Research, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio; Department of Surgery, The Ohio State University, Columbus, Ohio. Electronic address: david.brigstock@nationwidechildrens.org.
David R
DR
Brigstock
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D055354:Exosomes / Q000378:metabolism; D005786:Gene Expression Regulation; D055166:Hepatic Stellate Cells / Q000473:pathology / Q000502:physiology; D006801:Humans; D008103:Liver Cirrhosis / Q000209:etiology* / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D061251:Primary Cell Culture; D015398:Signal Transduction; D015854:Up-Regulation
D020413:3' Untranslated Regions; D035683:MicroRNAs; C534780:Mirn199 microRNA, mouse; C531408:mirn199 microRNA, human; D055513:Connective Tissue Growth Factor
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