N6-methyladenosine (m6A)-circHECA facilitates the differentiation of SHF stem cells into hair follicle lineage through miR-449a-5p/LEF1 mediated Wnt/β-catenin pathway in cashmere goats

OBJECTIVE: This study investigates the potential effect of N6-methyladenosine (m6A)- circHECA on the differentiation of SHF stem cells into hair follicle lineage along with the underlying molecular mechanisms in cashmere goats. METHODS: The effect of m6A-circHECA on the differentiation of SHF stem cells into hair follicle lineage was assessed through knockdown its expression in SHF stem cells derived from cashmere goats. The functional significance of m6A modification in circHECA functional exertion was confirmed through transfecting m6A deficient mutants into circHECA knockdown SHF stem cells. The competitive bindings of miR-449a-5p to m6A-circHECA and the 3´-untranslated region of LEF1 mRNA was investigated using Dualluciferase reporter assay. RESULTS: The m6A-circHECA exhibited significantly higher expression in SHF stem cells post-differentiation than pre-differentiation. Moreover, m6A-circHECA facilitated the differentiation process of SHF stem cells into hair follicle lineages. The m6A-circHECA sequestering miR-449a-5p, to enhance the expression of the LEF1 gene in SHF stem cells and activating the Wnt/β‑catenin signaling pathway. We further demonstrated that the m6A modification within circHECA is necessary for the miR-449a-5p/LEF1 mediated Wnt/β-catenin pathway, which promote the differentiation of SHF stem cells into hair follicle lineages via the introduction of a m6A-deffcient mutant of circHECA. CONCLUSION: The m6A-circHECA facilitates the differentiation of SHF stem cells into hair follicle lineage through miR-449a-5p/LEF1 mediated Wnt/β-catenin pathway in cashmere goats.