Resolution factor 15-epi-Lipoxin A4 modulates miRNA-499 induced differentiation of cardiosphere-derived stem cells through dual inhibition of Wnt/β-catenin and TGFβ/SMAD signalling axes

Abstract

Background: Cardiosphere derived stem cells (CDSCs) are signified as a valued source of stem cell therapeutics for treating a gamut of cardiovascular ailments. However, challenges in the cardiogenic differentiation process culminate in stem cell transplantation failure. Hence, it is imperative to understand the comprehensive mechanisms and the role of endogenous factors in cardiac differentiation. Objectives: Objective of the current study is to check the effect of 15-epi-lipoxin A4 (15E-LXA4)—a stable analogue of lipoxin A4, a pro-resolving agonist—on miR-499 (a cardiac-specific microRNA)-induced differentiation of CDSCs. Methods: CDSCs were transfected with lentiviral vectors bearing miR-499 and subjected to 15E-LXA4 treatment. Then, the treatment effects on surface markers (c-kit and CD105), cardiogenic gene markers (NKX2.5, GATA4, and cTnI), Wnt/β-catenin signaling (Wnt3a and phosphorylated/dephosphorylated β-catenin ratio) and TGFβ/SMAD signaling (TGFβ1 and SMAD3) were evaluated. Results: 15E-LXA4 treatment repressed miR-499 over-expression induced cardiac differentiation, manifested by enhanced expression of surface markers, Wnt/β-catenin and TGFβ/SMAD signaling parameters and reduced expression cardiogenic gene markers, plausibly through activation of estrogen receptor (ERα). Conclusions: These findings give reason to understand the role of endogenous resolution factors like 15E-LXA4, as it displayed reciprocal effect on miR-499 induced cardiac differentiation in CDSCs.

Keywords

• 15-epi-lipoxin A4
• miR-499
• Wnt3a
• TGFβ1
• SMAD3