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Empagliflozin and liraglutide ameliorate HFpEF in mice via augmenting the Erbb4 signaling pathway

Acta Pharmacologica Sinica (2024)Cite this article

Abstract

Heart failure with preserved ejection fraction (HFpEF) is closely associated with metabolic derangement. Sodium glucose cotransporter-2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1RA) exert anti-HFpEF effects, but the underlying mechanisms remain unclear. In this study, we explored the anti-HFpEF effects of empagliflozin and liraglutide and the underlying molecular mechanisms in a mouse model of HFpEF. This model was established by high-fat diet (HFD) feeding plus Nω-nitro-L-arginine methyl ester (L-NAME) treatment. The mice were treated with empagliflozin (20 mg·kg−1·d−1, i.g.) or liraglutide (0.3 mg·kg−1·d−1, i.p.) or their combination for 4 weeks. At the end of the experimental protocol, cardiac function was measured using ultrasound, then mice were euthanized and heart, liver, and kidney tissues were collected. Nuclei were isolated from frozen mouse ventricular tissue for single-nucleus RNA-sequencing (snRNA-seq). We showed that administration of empagliflozin or liraglutide alone or in combination significantly improved diastolic function, ameliorated cardiomyocyte hypertrophy and cardiac fibrosis, as well as exercise tolerance but no synergism was observed in the combination group. Furthermore, empagliflozin and/or liraglutide lowered body weight, improved glucose metabolism, lowered blood pressure, and improved liver and kidney function. After the withdrawal of empagliflozin or liraglutide for 1 week, these beneficial effects tended to diminish. The snRNA-seq analysis revealed a subcluster of myocytes, in which Erbb4 expression was down-regulated under HFpEF conditions, and restored by empagliflozin or liraglutide. Pseudo-time trajectory analysis and cell-to-cell communication studies confirmed that the Erbb4 pathway was a prominent pathway essential for both drug actions. In the HFpEF mouse model, both empagliflozin and liraglutide reversed Erbb4 down-regulation. In rat h9c2 cells, we showed that palmitic acid- or high glucose-induced changes in PKCα and/or ERK1/2 phosphorylation at least in part through Erbb4. Collectively, the single-cell atlas reveals the anti-HFpEF mechanism of empagliflozin and liraglutide, suggesting that Erbb4 pathway represents a new therapeutic target for HFpEF.

Effects and mechanisms of action of empagliflozin and liraglutide in HFpEF mice. HFpEF was induced with a high-fat diet and L-NAME for 15 weeks, and treatment with empagliflozin and liraglutide improved the HFpEF phenotype. Single nucleus RNA sequencing (snRNA-seq) was used to reveal the underlying mechanism of action of empagliflozin and liraglutide.

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Fig. 1: Empagliflozin and liraglutide improve heart function in HFpEF mice.
Fig. 2: Empagliflozin and/or liraglutide improve metabolism in HFpEF mice.
Fig. 3: Single cell sequencing landscape of HFpEF mice treated with empagliflozin/liraglutide.
Fig. 4: Erbb4-related signaling pathway is significantly enriched in the differentially expressed genes in myocyte subclusters caused by empagliflozin/liraglutide.
Fig. 5: Empagliflozin/liraglutide induces the heterogeneity of myocytes phenotypes and functions.
Fig. 6: Empagliflozin/liraglutide enhances the cell-to-cell communication between myocytes and other cell types.
Fig. 7: Empagliflozin/liraglutide regulates the Erbb4 signaling pathway in the heart.
Fig. 8: Erbb4 regulates ERK1/2 and PKCα phosphorylation in vitro.

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Data availability

The authors state that the main supporting data for the results of this study are available in the article and its supplementary information file. All sequencing data in this study are stored in the China National Center for Bioinformation/Beijing Institute of Genomics, Chinese Academy of Sciences (GSA: CRA012441) that are publicly accessible at https://ngdc.cncb.ac.cn/gsa. Additional data is available from corresponding authors upon request.

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Acknowledgements

This study was supported by grants from the National Key R&D Program of China (Grant No. 2021YFC2500500) and National Natural Science Foundation of China (No. 82003740, No. 82174031, No. 82070464, No. 82370444, No. 82102804), the Doctoral Research Fund (RC2021015), the China Postdoctoral Science Foundation (No. 2021M693102), and Scientific Research Project of Anhui Provincial Education Department (2022AH040184). This work was also supported by the Program for Innovative Research Team of The First Affiliated Hospital of USTC (CXGG02), Anhui Provincial Key Research and Development Program (Grant No. 202104j07020051), Anhui Provincial Natural Science Foundation (Grant No. 2208085J08). Graphical Abstract was created with Figdraw.

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  1. These authors contributed equally: Xia-yun Ni, Xiao-jun Feng, Zhi-hua Wang

Authors and Affiliations

  1. Department of Endocrinology, Institute of Endocrine and Metabolic Disease, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, Clinical Research Hospital of Chinese Academy of Sciences (Hefei), University of Science and Technology of China, Hefei, 230036, China

    Xia-yun Ni, Xiao-jun Feng, Zhi-hua Wang, Suo-wen Xu & Jian-ping Weng

  2. Department of Pharmacy, The First Affiliated Hospital of University of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China (USTC), Hefei, 230001, China

    Xia-yun Ni, Xiao-jun Feng, Yang Zhang & Li-qin Tang

  3. School of Pharmacy, University of Queensland, Pharmacy Australia Centre of Excellence, Woolloongabba, QLD, 4102, Australia

    Peter J. Little

  4. Division of Life Sciences and Medicine, University of Science and Technology of China (USTC), Hefei, 230022, China

    Yang Cao

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Contributions

XJF and XYN conducted experiments, analyzed data, and wrote papers. ZHW analyzed single-cell data and wrote the paper. YZ is involved in animal experiments. PJL and YC provided input on the pharmacology and therapeutics of the index drugs and edited and proofread the intermediate and the final versions of the paper. JPW, LQT, and SWX conceived projects, designed experiments, and revised papers.

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Correspondence to Suo-wen Xu, Li-qin Tang or Jian-ping Weng.

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Ni, Xy., Feng, Xj., Wang, Zh. et al. Empagliflozin and liraglutide ameliorate HFpEF in mice via augmenting the Erbb4 signaling pathway. Acta Pharmacol Sin (2024). https://doi.org/10.1038/s41401-024-01265-0

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