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Functional Association of miR-133b and miR-21 Through Novel Gene Targets ATG5, LRP6 and SGPP1 in Coronary Artery Disease

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Abstract

Background

Atherosclerosis, a progressive manifestation of coronary artery disease, has been observed to be regulated by microRNAs (miRNAs) targeting various protein-coding genes involved in several pathophysiological events of coronary artery disease.

Objective

In our previous report, we identified differential expression profiles of candidate miRNAs, miR-133b and miR-21, in patients with coronary artery disease as compared with controls, suggesting their possible implication in the pathophysiology of coronary artery disease. To better understand the functional role of these miRNAs, we sought to predict and validate their target genes while assessing the expression pattern of these genes in patients with coronary artery disease, as well as in macrophages.

Methods

Potential target genes of miR-133b and miR-21 were predicted bioinformatically followed by validation through the identification of their expression at  the protein level in patients with coronary artery disease (n-30), as well as in macrophages treated with respective miRNA inhibitors (antagomiRs), through immunoblotting.

Results

SGPP1, a gene associated with the sphingolipid pathway, was predicted to be a potential target gene of miR-133b while ATG5 and LRP6 were target genes of miR-21 while being associated with autophagy and Wnt signalling pathways, respectively. SGPP1 was observed to be upregulated significantly (p = 0.019) by 2.07-fold, whereas ATG5 and LRP6 were found to be downregulated (p = 0.026 and 0.007, respectively) by 3.28-fold and 8.46-fold, respectively, in patients with coronary artery disease as compared with controls. Expression patterns of all the genes were also found to be modulated when cells were treated with respective miRNA inhibitors.

Conclusions

Results from the present study suggest that SGPP1, ATG5 and LRP6, target genes of miR-133b and miR-21, may serve as potential therapeutic hotspots in the management of coronary artery disease, which undoubtedly merit further experimental confirmation.

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Acknowledgements

We acknowledge the infrastructural support to Kamna Srivastava and Daman Saluja from ACBR, University of Delhi and the Department of Science and Technology grant to Kamna Srivastava. Dinesh Kumar gratefully acknowledges University Grant Commission/Council of Scientific and Industrial Research, India for providing the JRF/SRF fellowship.

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Authors and Affiliations

  1. Dr. B R Ambedkar Center for Biomedical Research, University of Delhi, New Delhi, 110007, India

    Dinesh Kumar, Daman Saluja & Kamna Srivastava

  2. Department of Cardiology, All India Institute of Medical Sciences, New Delhi, 110029, India

    Rajiv Narang

  3. Delhi School of Public Health, IoE, University of Delhi, New Delhi, India

    Daman Saluja

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  1. Dinesh Kumar
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  2. Rajiv Narang
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Correspondence to Kamna Srivastava.

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Funding

This research was supported by the following grants: infrastructure facility from the Department of Science and Technology, WOS-A/LS-633/2017 granted to Kamna Srivastava and the University Grant Commission/Council of Scientific and Industrial Research UGC/CSIR-JRF/SRF fellowship (Ref. No. 22/12/2013(ii)EU-V) granted to Dinesh Kumar.

Conflict of interest

Dinesh Kumar, Rajiv Narang, Daman Saluja and Kamna Srivastava have no conflicts of interest that are directly relevant to the content of this article.

Ethics approval

The study protocol for human samples was approved by the Human Ethics Committee, AIIMS, New Delhi (IEC/NP-207/2012 and RP-17/2012) as well as the Human Ethics Committee, ACBR, DU, Delhi (F.50-2/Eth.Com/ACBR/169/2014).

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A written and signed informed consent form was obtained from all the participants of this study.

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The investigators have adhered to controlled-access publication and obtained consent to publish the study results.

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The data that support the findings of this study are available on reasonable request from the corresponding author.

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Authors' Contributions

DK and KS initiated the study, contributed to its conception and design, and drafted the manuscript. KS contributed to the project administration. DK and KS performed the experimental investigation. RN contributed to the screening and recruitment of the study subjects as well as clinical sample analysis. KS, DS, and DK did the formal analysis of the data. KS, DS, and DK reviewed and edited the manuscript. KS and DS supervised the research process. All authors have read and approved the publication of the manuscript.

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Kumar, D., Narang, R., Saluja, D. et al. Functional Association of miR-133b and miR-21 Through Novel Gene Targets ATG5, LRP6 and SGPP1 in Coronary Artery Disease. Mol Diagn Ther 26, 655–664 (2022). https://doi.org/10.1007/s40291-022-00615-0

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