LncRNA H19/miR-107 regulates endothelial progenitor cell pyroptosis and promotes flow recovery of lower extremity ischemia through targeting FADD

• Published in: Biochimica et biophysica acta. Molecular basis of disease Vol. 1870; no. 7; p. 167323
• Main Authors: Huang, Lin, Ye, Yanchen, Sun, Yunhao, Zhou, Zhihao, Deng, Tang, Liu, Yunyan, Wu, Ridong, Wang, Kangjie, Yao, Chen
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.10.2024
• Subjects: Animals; Cell Movement - genetics; Cell Proliferation; ceRNA; Critical limb ischemia; Disease Models, Animal; Endothelial progenitor cell therapy; Endothelial Progenitor Cells - metabolism; Fas-Associated Death Domain Protein - genetics; Fas-Associated Death Domain Protein - metabolism; Humans; Ischemia - genetics; Ischemia - metabolism; Ischemia - pathology; lncRNA H19; Lower Extremity - blood supply; Lower Extremity - pathology; Male; Mice; Mice, Inbred C57BL; MicroRNAs - genetics; MicroRNAs - metabolism; Neovascularization, Physiologic - genetics; Peripheral Arterial Disease - genetics; Peripheral Arterial Disease - metabolism; Peripheral Arterial Disease - pathology; Pyroptosis; Pyroptosis - genetics; RNA, Long Noncoding - genetics; RNA, Long Noncoding - metabolism; lncRNA H19; ceRNA; Pyroptosis; Critical limb ischemia; Endothelial progenitor cell therapy
• ISSN: 0925-4439; 1879-260X; 1879-260X
• DOI: 10.1016/j.bbadis.2024.167323

Peripheral artery disease (PAD) is an ischemic disease with a rising incidence worldwide. The lncRNA H19 (H19) is enriched in endothelial progenitor cells (EPCs), and transplantation of pyroptosis-resistant H19-overexpressed EPCs (oe-H19-EPCs) may promote vasculogenesis and blood flow recovery in PAD, especially with critical limb ischemia (CLI). EPCs isolated from human peripheral blood was characterized using immunofluorescence and flow cytometry. Cell proliferation was determined with CCK8 and EdU assays. Cell migration was assessed by Transwell and wound healing assays. The angiogenic potential was evaluated using tube formation assay. The pyroptosis pathway-related protein in EPCs was detected by western blot. The binding sites of H19 and FADD on miR-107 were analyzed using Luciferase assays. In vivo, oe-H19-EPCs were transplanted into a mouse ischemic limb model, and blood flow was detected by laser Doppler imaging. The transcriptional landscape behind the therapeutic effects of oe-H19-EPCs on ischemic limbs were examined with whole transcriptome sequencing. Overexpression of H19 in EPCs led to an increase in proliferation, migration, and tube formation abilities. These effects were mediated through pyroptosis pathway, which is regulated by the H19/miR-107/FADD axis. Transplantation of oe-H19-EPCs in a mouse ischemic limb model promoted vasculogenesis and blood flow recovery. Whole transcriptome sequencing indicated significant activation of vasculogenesis pathway in the ischemic limbs following treatment with oe-H19-EPCs. Overexpression of H19 increases FADD level by competitively binding to miR-107, leading to enhanced proliferation, migration, vasculogenesis, and inhibition of pyroptosis in EPCs. These effects ultimately promote the recovery of blood flow in CLI. In vitro, the overexpression of H19 in EPCs increases the expression of the anti-pyroptosis protein FADD by targeting miR-107. This upregulation enhances the resistance of EPCs to pyroptosis, improving their viability in the ischemic environment of the lower limbs. Consequently, this augmentation of EPCs pyroptosis resistance promotes the recovery of blood flow in the lower limbs, ameliorating symptoms associated with lower limb ischemia. [Display omitted] •Overexpression of lncRNA H19 significantly enhances anti-pyroptosis capacity of EPCs.•Oe-H19-EPCs treatment promotes blood flow recovery and improves limb ischemic in mice.•Transplantation of oe-H19-EPCs to treat limb ischemic activates vasculogenesis pathway.