Cellular molecular mechanism of EndoMT and tube formation dysfunction in iPSC-ECs of pulmonary arterial hypertension / 基础医学与临床

ABSTRACT

Objective To investigate the phenotypic and functional differences of endothelial cells derived from in -duced pluripotent stem cells(iPSC-ECs)between HPAH patient(HPAH)and control donor(CON),and clarify the molecular mechanism of phenotypic changes in BMPRII deficient ECs which is a pathological characteristic of PAH.Methods To differentiate the iPSCs derived from human pulmonary arterial smooth muscle cells (hPASMCs)into ECs.The expression of several genes related to stem cell and endothelial marker were analyzed at different time points during the differentiation.Immunofluorescence staining showed the expression of surface mark-ers of iPSC-ECs.The transcription factors involved in the EndoMT process were detected by real -time quantitative PCR(qPCR).HPAH and CON-derived iPSC-ECs were compared for tube formation.VEGFR2 mRNA and protein expression were detected by qPCR and immunofluorescence staining.Results The expression of several endothelial cell related genes of HPAH were different from CON during differentiation through they both expressed pluripotency genes.The expressions of α-SMA,HMGA1,Slug and Snail1 in HPAH iPSC-ECs were significantly higher as com-pared with CON ECs(P<0.05).The reduced tube formation of HPAH-derived ECs could be rescued by VEGF165.Their VEGFR2 mRNA and protein expression were lower as compared with CON ECs.Conclusions Enhanced HMGA1,Slug and Snail1 involve in the EndoMT of iPSC-ECs from HAPH,reduced VEGFR2 may con-tribute to tube formation dysfunction in pulmonary vascular remodeling of PAH.