A smooth muscle cell lncRNA controls angiogenesis in chronic limb-threatening ischemia through a miR-143-3p/HHIP signaling axis

Peripheral artery disease (PAD), affecting over 200 million people globally, often advances to chronic limb-threatening ischemia (CLTI), resulting in severe complications such as limb amputation and heightened mortality. Despite the potential of therapeutic angiogenesis, the mechanisms of cell-cell communication and transcriptional changes driving PAD are not fully understood. Profiling long non-coding RNAs (lncRNAs) from human gastrocnemius muscles of human subjects with or without CLTI revealed that a vascular smooth muscle cell (SMC)-enriched lncRNA CARMN was reduced with CLTI. This study explored the role of how SMC-driven regulation of angiogenesis in limb ischemia is conferred via a unique lncRNA-miRNA coupled signaling pathway.To investigate this interesting project, we perform three RNA-seq project. The first one was performed between hypoxia and normoxia VSMCs to mimic the HLI disease in patients. The second one was performed between CARMN KO SMC and CARMN WT SMC to find the potential downstream signaling pathway. The third one was about the RNA-seq of the gastro muscle obtained from CARMN KO and CARMN WT mice after HLI surgery. To explore this intriguing research question, we conducted three RNA-seq experiments. The first experiment compared VSMCs under hypoxic and normoxic conditions to simulate the HLI disease state observed in patients. The second experiment analyzed CARMN knockout (KO) SMCs versus CARMN wild-type (WT) SMCs to identify potential downstream signaling pathways. The third experiment involved RNA-seq analysis of gastrocnemius muscle tissue obtained from CARMN KO and CARMN WT mice following HLI surgery.