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Single Cell Genomics Identifies Unique Cardioprotective Phenotype of Stem Cells derived from Epicardial Adipose Tissue under Ischemia

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Abstract

The conventional management strategies of myocardial infarction (MI) are effective to sustain life; however, myocardial regeneration has not been achieved owing to the inherently poor regenerative capacity of the native myocardium. Stem cell-based therapies are promising; however, lineage specificity and undesired differentiation profile are challenging. Herein, we focused on the epicardial fat (EF) as an ideal source for mesenchymal stem cells (MSCs) owing to the proximity and same microvasculature with cardiac muscle. Unfortunately, the epicardial adipose tissue derived stem cells (EATDS) remain understudied regarding their phenotype heterogeneity and cardiac regeneration potential. As EF closely reflects the cardiac pathology during ischemia, the present study aims to determine the EATDS subpopulations under simulated ischemic and reperfused conditions employing single cell RNA sequencing (scRNAseq). EATDS were isolated from three hyperlipidemic Yucatan microswine and were divided into Control, Ischemia (ISC), and Ischemia/reperfusion (ISC/R). The scRNAseq analysis was performed using 10 genomics platform which revealed 18 unique cell clusters suggesting the existence of heterogeneous phenotypes. The upregulated genes were taken into consideration and subsequent functional assessment revealed the cardioprotective phenotypes with diverse mechanisms including epigenetic regulation (Cluster 1), myocardial homeostasis (Cluster 1), cell integrity and cell cycle (Clusters 2 and 3), prevention of fibroblast differentiation (Cluster 4), differentiation to myocardial lineage (Cluster 6), anti-inflammatory responses (Clusters 5, 8, and 11), prevention of ER-stress (Cluster 9), and increasing the energy metabolism (Cluster 10). These unique phenotypes of heterogeneous EATDS population open significant translational opportunities for myocardial regeneration and cardiac management.

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Data with the raw counts matrices and annotation are available upon request from the authors through proper channels.

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Acknowledgements

Authors are grateful to Dr. Hung Long, SC2 Core—Children Hospital Los Angeles, and Dr Anja Bastin and Dr. Elizabeth Collins, 10x genomics, for their technical support in multiple aspects of scRNAseq analysis.

Funding

The research work of FT is supported by the startup funds from WU and DK Agrawal is supported by research grants R01 HL144125 and R01HL147662 from the National Institutes of Health, USA. The content of this original article is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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  1. Department of Translational Research, Western University of Health Sciences, 309 E. Second Street, Pomona, CA, 91766-1854, USA

    Finosh G. Thankam & Devendra K. Agrawal

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  1. Finosh G. Thankam
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FT – conceptualization, experiments, data generation and analysis, manuscript preparation, and manuscript edits. DK—conceptualization, data generation and analysis, manuscript preparation, and manuscript edits.

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Correspondence to Finosh G. Thankam.

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Thankam, F.G., Agrawal, D.K. Single Cell Genomics Identifies Unique Cardioprotective Phenotype of Stem Cells derived from Epicardial Adipose Tissue under Ischemia. Stem Cell Rev and Rep 18, 294–335 (2022). https://doi.org/10.1007/s12015-021-10273-0

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