TGFß regulates metabolism of human skeletal muscle cells by miRNAs

S Höckele; P Huypens; C Hoffmann; T Jeske; M Hastreiter; A Böhm; J Beckers; HU Häring; M Hrabe de Angelis; C Weigert

doi:10.1055/s-0038-1641809

Diabetologie und Stoffwechsel, Table of Contents

Diabetologie und Stoffwechsel 2018; 13(S 01): S19
DOI: 10.1055/s-0038-1641809

Poster

Typ-2-Diabetes – Pathophysiologie I

Georg Thieme Verlag KG Stuttgart · New York

TGFß regulates metabolism of human skeletal muscle cells by miRNAs

S Höckele

¹Helmholtz Zentrum München, Institute of Experimental Genetics, Neuherberg, Germany

²German Center for Diabetes Research (DZD), Neuherberg, Germany

,

P Huypens

¹Helmholtz Zentrum München, Institute of Experimental Genetics, Neuherberg, Germany

,

C Hoffmann

³University Hospital Tübingen, Division of Endocrinology, Diabetology, Angiology, Nephrology, Pathobiochemistry and Clinical Chemistry, Department of Internal Medicine IV, Tübingen, Germany

,

T Jeske

⁴Helmholtz Zentrum München, Institute of Bioinformatics and Systems Biology, Neuherberg, Germany

⁵Dr. Hauner Children's Hospital, Ludwig-Maximilians-Universität München, Department of Pediatrics, München, Germany

,

M Hastreiter

⁶Helmholtz Zentrum München, Institute of Computational Biology, Neuherberg, Germany

⁷Technische Universität München, Lehrstuhl für Genomorientierte Bioinformatik, Freising, Germany

,

A Böhm

²German Center for Diabetes Research (DZD), Neuherberg, Germany

³University Hospital Tübingen, Division of Endocrinology, Diabetology, Angiology, Nephrology, Pathobiochemistry and Clinical Chemistry, Department of Internal Medicine IV, Tübingen, Germany

⁸University of Tübingen, Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum München, Tübingen, Germany

,

J Beckers

¹Helmholtz Zentrum München, Institute of Experimental Genetics, Neuherberg, Germany

²German Center for Diabetes Research (DZD), Neuherberg, Germany

⁹Technische Universität München, Institute of Experimental Genetics, Freising, Germany

,

HU Häring

²German Center for Diabetes Research (DZD), Neuherberg, Germany

³University Hospital Tübingen, Division of Endocrinology, Diabetology, Angiology, Nephrology, Pathobiochemistry and Clinical Chemistry, Department of Internal Medicine IV, Tübingen, Germany

⁸University of Tübingen, Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum München, Tübingen, Germany

,

M Hrabe de Angelis

¹Helmholtz Zentrum München, Institute of Experimental Genetics, Neuherberg, Germany

²German Center for Diabetes Research (DZD), Neuherberg, Germany

⁹Technische Universität München, Institute of Experimental Genetics, Freising, Germany

,

C Weigert

²German Center for Diabetes Research (DZD), Neuherberg, Germany

³University Hospital Tübingen, Division of Endocrinology, Diabetology, Angiology, Nephrology, Pathobiochemistry and Clinical Chemistry, Department of Internal Medicine IV, Tübingen, Germany

⁸University of Tübingen, Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum München, Tübingen, Germany

› Author Affiliations

Abstract

Full Text

TGFβ plays an important role in skeletal muscle by inhibiting cell differentiation and regulating inflammation and extracellular matrix production. Recent data link TGFβ with reduced expression of mitochondrial regulators, key regulators of β-oxidation, and negative influence on insulin response, providing a potential mechanism for exercise non-response.

To investigate the (post)transcriptional mechanisms of this TGFβ-dependent regulation and to identify epigenetic markers of exercise non-response we performed long and small RNA sequencing of primary human skeletal muscle cells stimulated with TGFβ1 before 5 d of differentiation. Sequencing data were analyzed bioinformatically using an annotation-based pipeline and USB (unique sequence based) analysis for sncRNAs.

Pathway and regulator enrichment analyses revealed downregulation of several transcripts of mitochondrial genes including all mitochondrially encoded subunits of complex I and IV after TGFβ1 stimulation. There was a decrease of transcripts of regulators of metabolic pathways, and of muscle cell differentiation while transcripts of extracellular matrix proteins were increased in TGFβ1 stimulated cells. Part of the data was confirmed on protein level using immunoblot analyses. Analysis of sncRNA sequencing data showed upregulation of miR-143 and miR-31 after TGFβ1 stimulation, and downregulation of miR-206, miR-133b, miR-499, and miR-208b. These results were confirmed by qPCR. A clustered set of sncRNAs is predicted to target within the mitochondrial genome.

Functional relevance of interesting sncRNAs will be validated in skeletal muscle cell experiments. Finally, we plan to investigate experimentally confirmed candidates in human muscle biopsy samples of individuals with a different response to exercise intervention.