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Interleukin-35 Mitigates ox-LDL-Induced Proatherogenic Effects via Modulating miRNAs Associated with Coronary Artery Disease (CAD)

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Abstract

Purpose

Recent emergence of miRNAs as important regulators of processes involving lesion formation and regression has highlighted miRNAs as potent therapeutic targets for the treatment of atherosclerosis. Few studies have reported the atheroprotective role of IL-35, a novel immunosuppressive and anti-inflammatory cytokine; however, miRNA-dependent regulation underlying the anti-atherosclerotic potential of IL-35 remains elusive.

Methods

THP-1 macrophages were incubated with human recombinant IL-35 (rIL-35) either in the presence or absence of ox-LDL. qRT-PCR was conducted to validate the expression levels of previously identified miRNAs including miR-197-5p, miR-4442, miR-324-3p, miR-6879-5p, and miR-6069 that were differentially expressed in peripheral blood mononuclear cells of coronary artery disease (CAD) patients vs. controls. Additionally, bioinformatic analysis was performed to predict miRNA-associated targets and their corresponding functional significance in CAD.

Results

Exogenous IL-35 significantly decreased the average area of ox-LDL-stimulated macrophages, indicating the inhibitory effect of IL-35 on lipid-laden foam cell formation. Furthermore, rIL-35 treatment alleviated the ox-LDL-mediated atherogenic effects by modulating the expression levels of aforementioned CAD-associated miRNAs in the cultured macrophages. Moreover, functional enrichment analysis of these miRNA-related targets revealed their role in the molecular processes affecting different stages of atheroslerotic plaque development, such as macrophage polarization, T cell suppression, lipoprotein metabolism, foam cell formation, and iNOS-mediated inflammation.

Conclusion

Our observations uncover the novel role of IL-35 as an epigenetic modifier as it influences the expression level of miRNAs implicated in the pathogenesis of atherosclerosis. Thus, IL-35 cytokine therapy–mediated miRNA targeting could be an effective therapeutic strategy against the development of early atheromas in asymptomatic high-risk CAD patients.

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Availability of Data and Materials

The data used to support the findings of this study are included within the article.

Abbreviations

rIL-35:

Recombinant interleukin-35

miRNA:

MicroRNA

Ox-LDL:

Oxidized low-density lipoprotein

CAD:

Coronary artery disease

PBMCs:

Peripheral blood mononuclear cells

VSMC:

Vascular smooth muscle cell

EC:

Endothelial cell

Treg:

Regulatory T cells

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Acknowledgements

We would like to acknowledge Mrs. Sakshi Mehta for providing her help in the basic cell culture work.

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Authors and Affiliations

  1. Department of Endocrinology, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, India

    Shipra Bhansali

  2. Department of Experimental Medicine and Biotechnology, Research Block-B, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, 160012, India

    Shipra Bhansali, Amit Kumar Yadav, Chetan Bakshi & Veena Dhawan

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Contributions

SB made intellectual contributions to the analysis of data, interpretation of the results, writing of the manuscript, and designing of figures. AKY collected the data and performed the experiments with CB. AKY performed the bioinformatics analysis with inputs from SB. CB made contributions to the conception and design of the work. VD supervised the project.

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Correspondence to Veena Dhawan.

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Bhansali, S., Yadav, A.K., Bakshi, C. et al. Interleukin-35 Mitigates ox-LDL-Induced Proatherogenic Effects via Modulating miRNAs Associated with Coronary Artery Disease (CAD). Cardiovasc Drugs Ther 37, 667–682 (2023). https://doi.org/10.1007/s10557-022-07335-x

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