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Adipose circular RNAs exhibit dynamic regulation in obesity and functional role in adipogenesis

Nature Metabolism volume 1, pages 688–703 (2019)Cite this article

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Abstract

Non-coding RNAs are emerging as novel regulators in adipocyte differentiation and function. Circular RNAs (circRNAs) are a new class of non-coding transcripts generated across all eukaryotic tissues, but their function in adipose biology remains unknown. Here we perform deep sequencing of visceral and subcutaneous fat to discover thousands of adipose circRNAs, many of which are species conserved, tissue specific and dynamically regulated during adipogenesis and obesity. We identify circTshz2-1 and circArhgap5-2 as indispensable regulators of adipogenesis in vitro. To characterize the function of circRNAs in vivo, we deliver adenoviral shRNA targeting circArhgap5-2 into mouse inguinal tissue and show that the expression of this circRNA is essential in maintaining the global adipocyte transcriptional programme involved in lipid biosynthesis and metabolism. We also demonstrate that the pro-adipogenic function of circArhgap5-2 is conserved in human adipocytes. Our results provide important evidence that circRNAs serve as important regulators in adipocyte differentiation and metabolism.

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Fig. 1: Global expression analysis of circular RNAs.
Fig. 2: Validation, tissue specificity and cellular localization of adipose circRNAs.
Fig. 3: CircRNA expression during adipogenesis and obesity.
Fig. 4: Knockdown of circTshz2-1 inhibits adipogenesis.
Fig. 5: Knockdown of circArhgap5-2 inhibits adipogenesis.
Fig. 6: circArhgap5-2 maintains the global adipogenic gene programme.
Fig. 7: In vivo knockdown of circArhgap5-2 downregulates adipogenic genes.
Fig. 8: circARHGAP5-1 function is conserved in human adipocytes.

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Data availability

The RNA-seq data generated in this study to build a database of adipose circRNAs in human and mouse adipose tissue, and to assess the global transcriptional impact of circArhgap5-2 knockdown in vivo and in vitro, are available in the NCBI Gene Expression Omnibus (GEO) database (Accession no: GSE118762). All other data generated and/or analysed to support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by Tanoto Initiative in Diabetes Research (to L.S.), National Medical Research Council’s Cooperative Basic Research Grant (NMRC/CBRG/0101/2016), Open Fund-Individual Research Grant (NMRC/OFIRG/0062/2017) and Ministry of Education Tier 2 grant (MOE2017-T2-2-015). Support was also provided by RNA Biology Center at CSI Singapore, NUS, from funding by the Singapore Ministry of Education’s Tier 3 grants, grant number MOE2014-T3-1-006. We thank S. Y. Chia, B. Pan, Z. Tiang and H. Hamadee for their technical assistance, Y.-H. Tseng (Joslin Diabetes Center) for providing cell lines and B. Lim and J. Li who offered their insight and expertise for this research.

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  1. These authors contributed equally: Camille Arcinas, Wilson Tan, Wenning Fang, Tresha P. Desai.

Authors and Affiliations

  1. Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore, Singapore

    Camille Arcinas, Wenning Fang, Tresha P. Desai, Diana Chee Siang Teh, Ufuk Degirmenci, Dan Xu & Lei Sun

  2. Genome Institute of Singapore, Agency for Science, Technology and Research, Singapore, Singapore

    Wilson Tan & Roger Foo

  3. Cardiovascular Research Institute, National University Health System, Singapore, Singapore

    Wilson Tan & Roger Foo

  4. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore, Singapore

    Lei Sun

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Contributions

L.S. and R.F. conceived and designed the study and acquired funds. C.A., W.T., D.X. and W.F. performed data analysis and developed and implemented the methodology. Data generation, curation and laboratory experiments were performed by C.A., W.T., W.F., T.P.D., D.T.C.S., U.D. and D.X. L.S., R.F., C.A. and W.T. wrote the manuscript and all authors contributed, read and approved the final version.

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Correspondence to Dan Xu, Roger Foo or Lei Sun.

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Arcinas, C., Tan, W., Fang, W. et al. Adipose circular RNAs exhibit dynamic regulation in obesity and functional role in adipogenesis. Nat Metab 1, 688–703 (2019). https://doi.org/10.1038/s42255-019-0078-z

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