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Targeting BCAA metabolism to potentiate metformin’s therapeutic efficacy in the treatment of diabetes in mice

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Abstract

Aims/hypothesis

An increasing body of evidence has shown that the catabolism of branched-chain amino acids (BCAAs; leucine, isoleucine and valine) is impaired in obese animals and humans, contributing to the development of insulin resistance and type 2 diabetes. Promoting BCAA catabolism benefits glycaemic control. It remains unclear whether BCAA catabolism plays a role in the therapeutic efficacy of currently used glucose-lowering drugs such as metformin.

Methods

Mice were treated with vehicle or metformin (250 mg/kg per day) for more than 4 weeks to investigate the effects of metformin in vivo. In vitro, primary mouse hepatocytes and HepG2 cells were treated with 2 mmol/l metformin. The therapeutic efficacy of metformin in the treatment of type 2 diabetes was assessed in genetically obese (ob/ob) mice and high-fat-diet-induced obese (DIO) mice. Enhancing BCAA catabolism was achieved with a pharmacological agent, 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid (BT2). The ob/ob mice were treated with a low-BCAA diet or intermittent protein restriction (IPR) to reduce BCAA nutritional intake.

Results

Metformin unexpectedly inhibited the catabolism of BCAAs in obese mice, resulting in an elevation of BCAA abundance. AMP-activated protein kinase (AMPK) mediated the impact of metformin on BCAA catabolism in hepatocytes. Importantly, enhancing BCAA catabolism via a pharmacological agent BT2 significantly potentiated the glucose-lowering effect of metformin while decreasing circulating BCAA levels in ob/ob and DIO mice. Similar outcomes were achieved by a nutritional approach of reducing BCAA intake. IPR also effectively reduced the circulating BCAA abundance and enhanced metformin’s glucose-lowering effect in ob/ob mice. BT2 and IPR treatments reduced the expression of fructose-1,6-bisphosphatase 1, a rate-limiting enzyme in gluconeogenesis, in the kidney but not liver, indicating the involvement of renal gluconeogenesis.

Conclusions/interpretation

Metformin self-limits its therapeutic efficacy in the treatment of type 2 diabetes by triggering the suppression of BCAA catabolism. Enhancing BCAA catabolism pharmacologically or reducing BCAA intake nutritionally potentiates the glucose-lowering effect of metformin. These data highlight the nutritional impact of protein on metformin’s therapeutic efficacy and provide new strategies targeting BCAA metabolism to improve metformin’s effects on the clinical outcome in diabetes.

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Abbreviations

AICAR:

5-Aminoinidazole 4-carboxamide 1-β-d-ribofuranoside

AMPK:

AMP-activated protein kinase

BCAA:

Branched-chain amino acid

BCAT:

Branched-chain aminotransferase

BCKA:

Branched-chain keto acid

BCKD:

BCKA dehydrogenase

BCKDK:

BCKD kinase

BCKDK-LKO:

Liver-specific Bckdk-knockout

BT2:

3,6-Dichlorobenzo[b]thiophene-2-carboxylic acid

DIO:

High-fat-diet-induced obese

FBP1:

Fructose-1,6-bisphosphatase 1

HFD:

High-fat diet

IPR:

Intermittent protein restriction

L-BCAA:

Low-BCAA

LPD:

Low-protein diet

NPD:

Normal protein diet

PP2Cm:

Protein phosphatase 2Cm

RT-qPCR:

Quantitative RT-PCR

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Author notes

  1. Xiaoyun Zhao and Xuejiao Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Immunology, Key Laboratory of Immune Microenvironment and Disease of the Educational Ministry of China, NHC Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin, China

    Xiaoyun Zhao, Xuejiao Zhang, Jingjing Pei, Yajin Liu, Wenyan Niu & Haipeng Sun

  2. Center for Cardiovascular Diseases, The Province and Ministry Co-Sponsored Collaborative Innovation Center for Medical Epigenetics, Tianjin Medical University, Tianjin, China

    Xuejiao Zhang & Haipeng Sun

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  1. Xiaoyun Zhao
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Corresponding authors

Correspondence to Wenyan Niu or Haipeng Sun.

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Data availability

Data generated in this study are available from the corresponding author upon request.

Funding

This work was supported by the National Key Research and Development Program of China (2019YFA0802503), the National Natural Science Foundation of China (92057107, 81570717, 31900819, 32200965, 82270856), the Collaborative Innovation Program of Shanghai Municipal Health Commission (2020CXJQ01) and the Tianjin Key Medical Discipline (Specialty) Construction Project (TJYXZDXK-032A).

Authors’ relationships and activities

HS participated in an advisory board for Ramino Bio Ltd. The authors declare that there are no other relationships or activities that might bias, or be perceived to bias, their work.

Contribution statement

HS and WN designed the study. XZhao, XZhang, JP and YL performed the experiments, contributed to data analysis and drafted the manuscript. HS and WN reviewed and edited the manuscript. All authors have read and agreed to the published version of the manuscript. HS and WN are responsible for the integrity of the work as a whole.

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Zhao, X., Zhang, X., Pei, J. et al. Targeting BCAA metabolism to potentiate metformin’s therapeutic efficacy in the treatment of diabetes in mice. Diabetologia 66, 2139–2153 (2023). https://doi.org/10.1007/s00125-023-05985-6

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