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LncRNA DLEU2 regulates sirtuins and mitochondrial respiratory chain complex IV: a novel pathway in obesity and offspring’s health

Abstract

Background

Long non-coding RNAs (lncRNAs) have emerged as a rapidly expanding area of interest in chronic diseases. They are mostly unknown for roles in metabolic regulation. Sirtuins, an epigenetic modulator class, regulate metabolic pathways. However, how sirtuins are regulated via lncRNA is unknown. We hypothesized that a high-fat high-fructose diet (HFD-HF) during pregnancy would increase the risk for obesity via lncRNA-Sirtuin pathways.

Methods

Female C57Bl/6 mice (F0) were fed either chow diet (CD) or HFD-HF for 6 weeks till birth. The pups (F1) were fed either CD or HFD-HF for 20 weeks. Expression of Dleu2, sirtuins, mitochondrial respiratory complexes, and oxidative stress were investigated in the F1 livers. Fasting blood glucose, insulin sensitivity, glucose tolerance, body and tissues weight were measured. A mechanistic interaction was then carried out using a DLEU2 knockdown experiment in the HepG2 cell.

Results

Dleu2 and sirtuins were both significantly decreased in the livers of HFD-HF fed male F1 whose mothers were either fed CD or HFD-HF during reproductive and pregnancy windows. Confirming this connection, upon silencing DLEU2, transcription levels of SIRT1 through 6 and translational levels of SIRT1, 3, 5, and 6 were significantly downregulated. Knockdown of DLEU2 significantly decreased the protein level of cytochrome-c oxidase (complex IV, MTCO1) without altering other mitochondrial complexes, decreased mitochondrial membrane potential, decreased ATP, and increased reactive oxygen species. Interestingly, in F1 livers, the protein level of MTCO1 was also significantly decreased under an HFD-HF diet or even under chow diet if the mother was exposed to HFD-HF.

Conclusion

Our findings reveal for the first time that one lncRNA can regulate sirtuins and a specific mitochondrial complex. Furthermore, diet or maternal diet can modulate Dleu2 and its downstream regulators in offspring, suggesting a potential role of DLEU2 in metabolic disorders over one or more generations.

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Fig. 1: Maternal diet effect on weight of body and liver, blood glucose, ITT, and GTT of F1 male mice on CD and HFD-HF.
Fig. 2: Maternal diet effect on gene expression of Dleu2 and Sirtuins in the livers of F1 male mice on CD and HFD-HF.
Fig. 3: Expressions of DLEU2, and Sirtuins in HepG2 after silencing DLEU2.
Fig. 4: Mitochondrial complex dysfunction in HepG2 after silencing of DLEU2 and maternal diet effect on protein level of MTCO1 in the livers of F1 male mice on CD and HFD-HF.
Fig. 5: Proposed model of HFD-HF leading to lncRNA DLEU2 regulated epigenetic circuit causing metabolic abnormalities in the liver.

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Acknowledgements

The work was supported by the Morris L Lichtenstein Jr. Medical Research Foundation. We would like to acknowledge Dr. Raktima Bhattacharya for running some qPCR plates. We also thank Faith Upton for helping with the model drawing and Nidhi Wunnava, Basel Kesbeh, Sarah Garcia, Martin Donjuan, Jazmin Diaz, Kayla Riley, Sumiya Wahab, Grace Chen, Jay Mendenhall, Crustal Chacko, Shroq Kesbeh, and Dr. Luis F Schutz for other activities (e.g., labeling, sac help etc).

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MC conceptualized the project and others carry out the experiments and data analysis. MC, MK, and JZ wrote the manuscript.

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Correspondence to Mahua Choudhury.

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Zhang, J., Kay, M.K., Park, M.H. et al. LncRNA DLEU2 regulates sirtuins and mitochondrial respiratory chain complex IV: a novel pathway in obesity and offspring’s health. Int J Obes 46, 969–976 (2022). https://doi.org/10.1038/s41366-022-01075-6

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