The Mechanism Of EIF3a Gene Mutation In The Process Ofleft Ventricular Noncompaction

Objective: To investigate the association between eIF3 a gene and LVNC,and explore potential mechanisms that the novel variant(c.1145A->G)in eIF3 a could lead to the development of LVNC.Methods: A novel eIF3 a variant,c.1145A->G,was identified by whole-exome sequencing in a familial pedigree with LVNC.Adenovirus vectors containing wild-type eIF3 a,the mutated version and vector version were constructed and co-infected into H9C2 cells.RT-PCR was used to detect the expression levels of eIF3 a mRNA after transfection.Cell proliferation,cell cycle and apoptosis were studied and were measured by proliferation assays,flow cytometry.Wound healing experiments were carried out to measure the rate of cell migration.The expression of vinculin was determined by immunofluorescence and western blot.Then,normal human induced pluripotent stem cells(hiPSCs)were further induced into cardiomyocytes,co-infected with adenovirus.Real-time PCR was used to detect the expression of stem cell markers and cardiac differential markers.Western blot was used to measured the phosphorylation of ERK1/2.Results: To Compared with the wild type group,the cell prolif eration capacity of the mutation group decreased significantly in the H9C2 cell(P<0.05).Flow cytometry show no difference in cell cycle,and apoptosis was significantly increased in the mutation group(P<0.05).The wound healing experiments showed that the cell migration abil ity was enhanced in mutation group(P<0.05).Confocal and Western bl ot also indicated enhanced expression of vinculin in mutation group(P<0.05).At the same time,compared with wild type,the expression of stem cell markers decreased in the mutation group,while the early cardiac differential markers NKx2.5 was high-expression(P<0.01),the late cardiac differential markers cTnT express lowly(P<0.01),the two groups showed opposite trend.The expression of phosphorylated erk1/2 decreased in the mutation group(P<0.01).Conclusion: The eIF3 a mutation inhibited the proliferation of H9C2 cells,induced apoptosis,promoted cell migration,and inhibited th e differentiation of human induced pluripotent stem cell-derived cardi omyocytes(hiPSC-CMs).The effect of the eIF3 a mutation may be at tributed to a decrease in expression of p-ERK1/2,which may caused decreased myocardium proliferation,differentiation,accelerated migratio n.This finding may provide some insight into the mechanism involved in LVNC development.