Long non-coding RNA TUG1 regulates smooth muscle cell differentiation via KLF4-Myocardin axis.

Abstract

Abdominal Aortic Aneurysms (AAAs) are asymptomatic vascular diseases that have life-threatening outcomes. Smooth-muscle cell (SMC) dysfunction plays an important role in AAA development. The contribution of non-coding genome, specifically the role of long non-coding RNAs (lncRNAs) in SMC dysfunction is relatively unexplored. we investigated the role of lncRNA TUG1 in SMC dysfunction. To identify potential lncRNAs relevant to SMC functionality, lncRNA profiling was performed in angiotensin-II treated SMCs. AAA was induced by angiotensin-II treatment in mice. Transcriptional regulation of TUG1 was studied using promoter luciferase and chromatin-immuno-precipitation experiments. Gain- or loss- of function experiments were performed in-vitro to investigate TUG1-mediated regulation of SMC function. Immunoprecipitation experiments were conducted to elucidate the mechanism underlying TUG1-mediated SMC dysfunction. TUG1 was upregulated in SMCs following angiotensin-II treatment. Similarly, TUG1 levels were elevated in abdominal aorta in a mouse model of angiotensin-II-induced AAA. Further investigations showed that angiotensin-II-induced TUG1 expression could be suppressed by inhibiting Notch-signaling pathway, both in-vitro and in mouse AAA model and that TUG1 is a direct transcriptional target of the Notch-pathway. In aneurysmal tissues, TUG1 expression was inversely correlated with the expression of SMC-contractile genes. Over-expression of TUG1 repressed SMC differentiation in-vitro, whereas siRNA/shRNA-mediated TUG1 knockdown showed an opposite effect. Mechanistically, TUG1 interacts with transcriptional repressor KLF4 and facilitates its recruitment to myocardin promoter ultimately leading to the repression of SMC-differentiation. In summary, our study uncovers a novel role for the lncRNA TUG1 wherein it modulates SMC-differentiation via the KLF4-Myocardin axis which may have potential implications in AAA development.