Autophagy-Related Activation of Hepatic Stellate Cells Reduces Cellular miR-29a by Promoting Its Vesicular Secretion

Xiaojie Yu; Natalia Elfimova; Marion Müller; Daniel Bachurski; Ulrike Koitzsch; Uta Drebber; Esther Mahabir; Hinrich P Hansen; Scott L Friedman; Sabine Klein; Hans Peter Dienes; Marianna Hösel; Reinhard Buettner; Jonel Trebicka; Vangelis Kondylis; Inge Mannaerts; Margarete Odenthal

doi:10.1016/j.jcmgh.2022.02.013

Autophagy-Related Activation of Hepatic Stellate Cells Reduces Cellular miR-29a by Promoting Its Vesicular Secretion

Cell Mol Gastroenterol Hepatol. 2022;13(6):1701-1716. doi: 10.1016/j.jcmgh.2022.02.013. Epub 2022 Feb 24.

Authors

Xiaojie Yu¹, Natalia Elfimova², Marion Müller¹, Daniel Bachurski³, Ulrike Koitzsch¹, Uta Drebber⁴, Esther Mahabir⁵, Hinrich P Hansen⁶, Scott L Friedman⁷, Sabine Klein⁸, Hans Peter Dienes², Marianna Hösel¹, Reinhard Buettner⁹, Jonel Trebicka¹⁰, Vangelis Kondylis¹¹, Inge Mannaerts¹², Margarete Odenthal¹³

Affiliations

¹ Institute for Pathology, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany.
² Institute for Pathology, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany.
³ Department I of Internal Medicine, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany.
⁴ Institute for Pathology, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany; Center of Integrative Oncology, University Clinic of Cologne and Bonn, Germany.
⁵ Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany.
⁶ Department I of Internal Medicine, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany; Center of Integrative Oncology, University Clinic of Cologne and Bonn, Germany.
⁷ Division of Liver Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, New York.
⁸ Department of Internal Medicine I, University Hospital Frankfurt, Frankfurt, Germany.
⁹ Institute for Pathology, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany; Center of Integrative Oncology, University Clinic of Cologne and Bonn, Germany.
¹⁰ Department of Internal Medicine I, University Hospital Frankfurt, Frankfurt, Germany; European Foundation for the Study of Chronic Liver Failure - EF CLIF, Barcelona, Spain.
¹¹ Institute for Pathology, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany; Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.
¹² Liver Cell Biology Research Group, Vrije Universiteit Brussel, Brussel, Belgium. Electronic address: [email protected].
¹³ Institute for Pathology, Medical Faculty and University Hospital of Cologne, University of Cologne, Cologne, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany; Center of Integrative Oncology, University Clinic of Cologne and Bonn, Germany. Electronic address: [email protected].

Abstract

Background & aims: Liver fibrosis arises from long-term chronic liver injury, accompanied by an accelerated wound healing response with interstitial accumulation of extracellular matrix (ECM). Activated hepatic stellate cells (HSC) are the main source for ECM production. MicroRNA29a (miR-29a) is a crucial antifibrotic miRNA that is repressed during fibrosis, resulting in up-regulation of collagen synthesis.

Methods: Intracellular and extracellular miRNA levels of primary and immortalized myofibroblastic HSC in response to profibrogenic stimulation by transforming growth factor β (TGFβ) or platelet-derived growth factor-BB (PDGF-BB) or upon inhibition of vesicular transport and autophagy processes were determined by quantitative polymerase chain reaction. Autophagy flux was studied by electron microscopy, flow cytometry, immunoblotting, and immunocytochemistry. Hepatic and serum miR-29a levels were quantified by using both liver tissue and serum samples from a cohort of chronic hepatitis C virus patients and a murine CCl₄ induced liver fibrosis model.

Results: In our study, we show that TGFβ and PDGF-BB resulted in decrease of intracellular miR-29a and a pronounced increase of vesicular miR-29a release into the supernatant. Strikingly, miR-29a vesicular release was accompanied by enhanced autophagic activity and up-regulation of the autophagy marker protein LC3. Moreover, autophagy inhibition strongly prevented miR-29a secretion and repressed its targets' expression such as Col1A1. Consistently, hepatic miR-29a loss and increased LC3 expression in myofibroblastic HSC were associated with increased serum miR-29a levels in CCl₄-treated murine liver fibrosis and specimens of hepatitis C virus patients with chronic liver disease.

Conclusions: We provide evidence that activation-associated autophagy in HSC induces release of miR-29a, whereas inhibition of autophagy represses fibrogenic gene expression in part through attenuated miR-29a secretion.

Keywords: Circulating MiRNA; Liver Fibrosis; MiR-29.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
Autophagy
Becaplermin / metabolism
Hepatic Stellate Cells / pathology
Hepatitis C, Chronic* / metabolism
Hepatitis C, Chronic* / pathology
Humans
Liver Cirrhosis / pathology
Mice
MicroRNAs / genetics*
MicroRNAs / metabolism
Transforming Growth Factor beta / metabolism

Substances

MIRN29 microRNA, mouse
MIRN29a microRNA, human
MicroRNAs
Transforming Growth Factor beta
Becaplermin

Grants and funding

R01 DK128289/DK/NIDDK NIH HHS/United States