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A microRNA screen reveals that elevated hepatic ectodysplasin A expression contributes to obesity-induced insulin resistance in skeletal muscle


Over 40% of microRNAs (miRNAs) are located in introns of protein-coding genes, and many of these intronic miRNAs are co-regulated with their host genes1,2. In such cases of co-regulation, the products of host genes and their intronic miRNAs can cooperate to coordinately regulate biologically important pathways3,4. Therefore, we screened intronic miRNAs dysregulated in the livers of mouse models of obesity to identify previously uncharacterized protein-coding host genes that may contribute to the pathogenesis of obesity-associated insulin resistance and type 2 diabetes mellitus. Our approach revealed that expression of both the gene encoding ectodysplasin A (Eda), the causal gene in X-linked hypohidrotic ectodermal dysplasia (XLHED)5, and its intronic miRNA, miR-676, was increased in the livers of obese mice. Moreover, hepatic EDA expression is increased in obese human subjects and reduced upon weight loss, and its hepatic expression correlates with systemic insulin resistance. We also found that reducing miR-676 expression in db/db mice increases the expression of proteins involved in fatty acid oxidation and reduces the expression of inflammatory signaling components in the liver. Further, we found that Eda expression in mouse liver is controlled via PPARγ and RXR-α, increases in circulation under conditions of obesity, and promotes JNK activation and inhibitory serine phosphorylation of IRS1 in skeletal muscle. In accordance with these findings, gain- and loss-of-function approaches reveal that liver-derived EDA regulates systemic glucose metabolism, suggesting that EDA is a hepatokine that can contribute to impaired skeletal muscle insulin sensitivity in obesity.

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Figure 1: Greater hepatic Eda and miR-676 expression in obese mice.
Figure 2: Increased hepatic EDA expression in human obesity.
Figure 3: PPARγ and RXR-α activity promotes Eda transcription in hepatocytes, which in turn can activate JNK and lead to Ser307 phosphorylation of IRS1 in skeletal muscle.
Figure 4: EDA-A2 overexpression in wild-type mice accelerates HFD-induced glucose intolerance, whereas Eda suppression in db/db mice ameliorates insulin resistance.

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We acknowledge J. Alber, B. Hampel, P. Scholl, N. Spenrath and D. Kutyniok for outstanding technical assistance. We greatly appreciate P. Schneider (Lausanne University) for providing Tabby mice and EDA expression vectors as well as outstanding support in performance of the EDA AlphaLISA assay. We acknowledge J. Wilson (University of Pennsylvania) for providing the pXR8 plasmid and J. Samulski (University of North Carolina) for providing the pXX6-80 plasmid. We acknowledge P. Frommolt for bioinformatics support and H. Büning for support of AAV production. This work was supported by a grant from the German Research Foundation (DFG) (BR 1492/7-1) to J.C.B., and we received funding from DFG within the framework of TRR134 and within the Excellence Initiative by German Federal and State Governments (CECAD). This work was funded (in part) by the Helmholtz Alliance (Imaging and Curing Environmental Metabolic Diseases, ICEMED) through the Initiative and Networking Fund of the Helmholtz Association. J.W.K. greatly appreciates funding from the Emmy Noether program of DFG (KO 4728/1-1). M.A. gratefully acknowledges support from a Manpei Suzuki Diabetes Foundation fellowship.

Author information




M.A. and J.C.B. designed the study and wrote the manuscript. M.A. and P.G. performed experiments, and M.A. analyzed data. E.T. performed the surgeries and with P.G. conducted the hyperinsulinemic–euglycemic clamp experiment. C.B. also supported the hyperinsulinemic–euglycemic clamp studies. H.N. and M.K. performed SILAC and proteomic analyses. P.J.A. contributed to AAV generation. J.S. provided critical reagents. J.A. performed miRNA sequencing, and S.M. analyzed the data. F.T.W. provided JNKSM-C mice. J.-W.K. provided the microarray data. M.B. provided the human samples and clinical data.

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Correspondence to Jens C Brüning.

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The authors declare no competing financial interests.

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Awazawa, M., Gabel, P., Tsaousidou, E. et al. A microRNA screen reveals that elevated hepatic ectodysplasin A expression contributes to obesity-induced insulin resistance in skeletal muscle. Nat Med 23, 1466–1473 (2017).

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