Circ-calm4 regulates hypoxia-induced pulmonary artery smooth muscle autophagy by binding Purb

Pulmonary hypertension (PH) is a serious and fatal disease characterized by pulmonary vasoconstriction and pulmonary vascular remodeling. The excessive autophagy of pulmonary artery smooth muscle cells (PASMCs) is one of the important factors of pulmonary vascular remodeling. A number of studies have shown that circular RNA (circRNA) can participate in the onset of PH. Our previous studies have shown that circRNA calmodulin 4 (circ-calm4) is involved in the progression of hypoxic PH. However, the role of circ-calm4 on regulation of hypoxic PH autophagy has not been reported. In this study, we demonstrated for the first time that hypoxia-mediated upregulated circ-calm4 expression has a key regulatory effect on autophagy in hypoxia-induced PASMCs and hypoxic PH mouse models. Knockdown of circ-calm4 both in vivo and in vitro can inhibit the autophagy in PASMCs induced by hypoxia. We also performed bioinformatics predictions and conducted ex-periments to verify that circ-calm4 bound to the purine-rich binding protein (Purb) to promote its expression in the nucleus, thereby initiating the transcription of autophagy-related protein Beclin1. Interestingly, we found that Beclin1 transcription initiated by Purb was accompanied by a modification of Beclin1 super-enhancer to improve transcription activity and efficiency. Overall, our results confirm that the circ-calm4/Purb/Beclin1 signal axis is involved in the occurrence of hypoxia-induced PASMCs autophagy, and the novel regulatory mechanisms and signals transduction pathways in PASMC autophagy induced by hypoxia.

Automated summary

Pulmonary hypertension (PH) is characterized by pulmonary vasoconstriction and vascular remodeling. Excessive autophagy of pulmonary artery smooth muscle cells (PASMCs) is an important factor in vascular remodeling. This study showed that hypoxia-induced circ-calm4 upregulation regulates autophagy in hypoxia-induced PASMCs. Knockdown of circ-calm4 inhibits hypoxia-induced autophagy in PASMCs. The circ-calm4/Purb/Beclin1 signal axis is involved in hypoxia-induced PASMCs autophagy.