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01-04-2020 | Obesity | Article

Obesity-induced reduced expression of the lncRNA ROIT impairs insulin transcription by downregulation of Nkx6.1 methylation

Authors: Fang Fang Zhang, Yu Hong Liu, Dan Wei Wang, Ting Sheng Liu, Yue Yang, Jia Min Guo, Yi Pan, Yan Feng Zhang, Hong Du, Ling Li, Liang Jin

Published in: Diabetologia | Issue 4/2020

Abstract

Aims/hypothesis

Although obesity is a predisposing factor for pancreatic beta cell dysfunction, the mechanisms underlying its negative effect on insulin-secreting cells is still poorly understood. The aim of this study was to identify islet long non-coding RNAs (lncRNAs) involved in obesity-mediated beta cell dysfunction.

Methods

RNA sequencing was performed to analyse the islets of high-fat diet (HFD)-fed mice and those of normal chow-fed mice (NCD). The function in beta cells of the selected lncRNA 1810019D21Rik (referred to in this paper as ROIT [regulator of insulin transcription]) was assessed after its overexpression or knockdown in MIN6 cells and primary islet cells, as well as in siRNA-treated mice. Then, RNA pull-down, RNA immunoprecipitation, coimmunoprecipitation and bisulphite sequencing were performed to investigate the mechanism of ROIT regulation of islet function.

Results

ROIT was dramatically downregulated in the islets of the obese mice, as well as in the sera of obese donors with type 2 diabetes, and was suppressed by HNF1B. Overexpression of ROIT in MIN6 cells and islets led to improved glucose homeostasis and insulin transcription. Investigation of the mechanism involved showed that ROIT bound to DNA methyltransferase 3a and caused its degradation through the ubiquitin proteasome pathway, which blocked the methylation of the Nkx6.1 promoter.

Conclusions/interpretation

These findings functionally suggest a novel link between obesity and beta cell dysfunction via ROIT. Elucidating a precise mechanism for the effect of obesity on lncRNA expression will broaden our understanding of the pathophysiological development of diabetes and facilitate the design of better tools for diabetes prevention and treatment.

Data availability

The raw RNA sequencing data are available from the NCBI Gene Expression Omnibus (GEO series accession number GSE139991).

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Title: Obesity-induced reduced expression of the lncRNA ROIT impairs insulin transcription by downregulation of Nkx6.1 methylation
Authors: Fang Fang Zhang
Yu Hong Liu
Dan Wei Wang
Ting Sheng Liu
Yue Yang
Jia Min Guo
Yi Pan
Yan Feng Zhang
Hong Du
Ling Li
Liang Jin
Publication date: 01-04-2020
Publisher: Springer Berlin Heidelberg
Keywords: Obesity
Obesity
Insulins
Insulins
Published in: Diabetologia / Issue 4/2020
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-020-05090-y

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