Abstract 18048: Dysregulated Microrna-30c and -376c Promotes Functional Disability of Diabetic Cardiac Stem Cells
Background: Diabetes mellitus leads to a progressive loss in the number and functional efficacy of cardiac stem cells (CSCs) and transplanted CSCs from diabetic heart failed to improve the recovery of damaged heart tissue. However, the underlying mechanism for this functional deficit remains unknown. Our recent study showed marked deregulation of microRNAs (miRs) in diabetic heart.
Objective: To investigate if deregulated miRs are the underlying molecular mechanism responsible for functional deterioration of diabetic CSCs.
Methods and Results: CSCs were isolated from type-2 diabetic and age-matched non-diabetic mice hearts (n=9 each) using collagenase digestion, followed by purification of the cells using stem cell specific antigen-1 (Sca-1) magnetic labelling. Following one week of culture, total RNA was extracted and miR expression profiles were analysed using an nCounter miR expression assay (Nanostring Technologies). Among 601 miRs evaluated, 14 miRs were significantly altered (12 up-regulated and 2 down-regulated, based on minimum of ±2 fold change and P<0.01) in diabetic CSCs. The online tool, ‘MiRPath’ narrowed this to 4 miRs (upregulation of miR-376c, miR-329 and miR-495 and downregulation of miR-30c) that are involved in regulation of stem cell proliferation and differentiation. Mass spectrometry analysis confirmed the downregulation of CDK6 and upregulation of VDAC1, target proteins for miR-376c and miR-30c respectively (P<0.05). Finally in vitro functional analysis using loss- or gain-of function in diabetic CSCs demonstrated marked improvement in the survival, proliferation and differentiation in diabetic CSCs only following the restoration of miR-30c and miR-376c (P<0.05 for all the parameters).
Conclusion: Therapeutic restoration of miR-30c and miR-376c improves the in vitro functional ability of diabetic CSCs.
Author Disclosures: R. Katare: Research Grant; Modest; Heart Foundation New Zealand. N. Purvis: None. S. Kumari: None.
- © 2016 by American Heart Association, Inc.