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Long non-coding RNA H19 inhibition promotes hyperglycemia in mice by upregulating hepatic FoxO1 levels and promoting gluconeogenesis

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Abstract

In a previous report from our laboratory, it was reported that hepatic levels of the long non-coding RNA (lncRNA), H19 are decreased in diabetic mice which elevates hepatic gluconeogenesis and glucose output. But, the mechanisms of H19 inhibition in elevating gluconeogenic genes’ transcription and promoting hepatic glucose output were not known. In this study, we aimed to decipher this regulatory role of H19 on glucose metabolism and on FoxO1, an important transcriptional regulator of gluconeogenesis. While H19 inhibition in HepG2 cells increased the levels of FoxO1, its overexpression led to significant inhibition in FoxO1 levels, thereby identifying H19 as an important regulator of FoxO1. Our data also demonstrates that in the absence of H19, there is increased occupancy of p53 on the FoxO1 promoter that possibly is responsible for increased FoxO1 transcription. In vivo silencing of H19 in normal mice caused hyperglycemia, hyperinsulinemia and impaired glucose, insulin, and pyruvate tolerance. Serum triglyceride and cholesterol levels, however, did not show any change. H19 inhibition significantly elevated the hepatic levels of FoxO1 and all the gluconeogenic genes. While fasting increased gluconeogenic genes’ transcription, the levels of H19 were decreased and these patterns reversed upon refeeding the mice. Thus, gluconeogenic genes and H19 levels show inverse patterns of expression, and these results indicate towards an important regulatory role of the lncRNA, H19. It acts as an upstream regulator of gluconeogenesis by regulating the transcription of FoxO1, an important transcription factor of gluconeogenic genes, and hence, regulates hepatic glucose metabolism.

Key messages

  • H19 regulates FoxO1 transcript and protein levels.

  • H19 inhibition increases p53 occupancy on the FoxO1 promoter that promotes FoxO1 transcription.

  • H19 inhibition in vivo induces hyperglycemia and impairs glucose, insulin, and pyruvate tolerance.

  • In vivo H19 inhibition increases the hepatic transcript levels of gluconeogenic genes and FoxO1.

  • Transcript levels of H19 and gluconeogenic genes are inversely regulated during fed and fasted states.

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Acknowledgments

The authors acknowledge CSIR (NG and DK) and UGC (ST), New Delhi, India, for the fellowships. The authors thank the team of the Animal House facility of CSIR-IGIB for the assistance during the in vivo experiments. We thank Mr. Ananda K Mukherjee, CSIR-Institute of Genomics and Integrative Biology for his help in the chromatin-immunoprecipitation experiments.

Funding

This work was supported by funding from the Council of Scientific and Industrial Research (CSIR), New Delhi, India (BSC0123).

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Authors and Affiliations

  1. CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, 110007, India

    Neha Goyal, Shweta Tiwary, Devesh Kesharwani & Malabika Datta

  2. CSIR-HRDC, Academy of Scientific and Innovative Research, Kamala Nehru Nagar, Ghaziabad, Uttar Pradesh, 201002, India

    Neha Goyal, Shweta Tiwary, Devesh Kesharwani & Malabika Datta

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  1. Neha Goyal
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Correspondence to Malabika Datta.

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Neha Goyal and Shweta Tiwary are co-first authors

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Goyal, N., Tiwary, S., Kesharwani, D. et al. Long non-coding RNA H19 inhibition promotes hyperglycemia in mice by upregulating hepatic FoxO1 levels and promoting gluconeogenesis. J Mol Med 97, 115–126 (2019). https://doi.org/10.1007/s00109-018-1718-6

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  • DOI: https://doi.org/10.1007/s00109-018-1718-6

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