Therapeutic Potential of miR-148a-3p Inhibition to Enhance Neovascularization and cellular senescence

https://doi.org/10.1007/s00392-024-02526-y

Fiene Daniel (Halle (Saale))1, L. Hehl (Halle (Saale))1, K. Kalies (Halle (Saale))1, T. D. Wils (Halle (Saale))1, J. Dutzmann (Halle (Saale))1, K. Knöpp (Halle (Saale))1, D. G. Sedding (Halle (Saale))1

1Universitätsklinikum Halle (Saale) Klinik und Poliklinik für Innere Medizin III Halle (Saale), Deutschland

 

To enhance healing after a myocardial infarction (MI), a well-coordinated angiogenic response is crucial. Current therapeutic approaches are insufficient. MicroRNAs (miRs) have been identified as regulators of angiogenesis and offer a promising strategy to improve neovascularization following MI. In this study, we investigate miR-148a-3p as a new miRNA candidate with therapeutic potential to enhance neovascularization post-MI, particularly in the context of cellular senescence.

The expression levels of miR-148a-3p were measured in Human Coronary Artery Endothelial Cells (HCAECs) and murine tissue using qPCR analysis. The effects of miR-148a-3p on cellular functions (proliferation, migration, cell death) were examined after pre- or anti-miR transfection in both replicative and non-replicative senescent HCAECs. Target gene and protein expressions were evaluated by qPCR and Western Blotting.

Initial screenings indicated that miR-148a-3p expression was upregulated in murine aortic endothelial cells of C57BL/6 mice (ages 3 vs. 24 months, p<0.05) and in a wire-induced vascular injury model (p<0.001). A similar effect was observed in vitro in senescent HCAECs (p<0.01), particularly under hypoxic conditions (p<0.05). Under growth conditions, miR-148a-3p levels were reduced compared to starvation, indicating a negative correlation with proliferation (p<0.05). Accordingly, downregulating miR-148a-3p improved proliferation in both non-senescent and senescent cells (p<0.01, p<0.001) and enhanced migration in senescent HCAECs (p<0.05). Systemic target research identified Akt1, NFKBIA, and RTN4 as potential targets, which were significantly upregulated following miR-148a-3p downregulation (p<0.001, p<0.01, p<0.05). The knockdown of miR-148a-3p reduced the expression of senescence markers such as p16ink4a, p21, p14arf, and CD44 (p<0.05). Immunofluorescence microscopy showed decreased nuclei size by anti-miR transfection as well as reduced b-galactosidase positive cells.

Our findings demonstrate that inhibiting miR-148a-3p enhances cellular functions, particularly in replicative senescent cells, strongly indicating that miRNA-148a-3p is involved in the angiogenic response and may serve as a therapeutic target. Future experiments will evaluate its potential in vivo using ultrasound-guided MI induction in mice.
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