背景:人类诱导多能干细胞(hiPSC)技术是产生患者特异性干细胞的宝贵工具,促进疾病建模,并调查疾病机制。然而,携带特定突变的iPSC可能由于某些固有特性而限制其临床应用。
目的:研究MERTK突变对hiPSCs的影响,并确定hiPSC来源的细胞外囊泡(EV)是否会影响异常细胞连接和分化潜能。
方法:我们采用非整合重编程技术来产生外周血来源的hiPSC,其具有和不具有MERTK突变的hiPSC。染色体核型分析,流式细胞术,和免疫荧光染色用于hiPSC鉴定。转录组学和蛋白质组学用于阐明与细胞连接异常和细胞分化潜能相关的表达模式。此外,从上清液中分离出电动汽车,并检查了它们的RNA和蛋白质货物,以研究hiPSC衍生的EV在干细胞连接和分化中的参与。
结果:生成的hiPSC,无论是否有MERTK突变,表现出正常的核型和表达的多能性标记;然而,具有MERTK突变的hiPSC表现出异常的粘附能力和分化潜能,通过转录组学和蛋白质组学分析证实。此外,hiPSC衍生的电动汽车参与各种生物过程,包括细胞连接和分化。
结论:具有MERTK突变的HiPSC显示出改变的连接特征和异常的分化潜能。此外,hiPSC衍生的电动汽车在各种生物过程中发挥了调节作用,包括细胞连接和分化。
BACKGROUND: Human induced pluripotent stem cell (hiPSC) technology is a valuable tool for generating patient-specific stem cells, facilitating disease modeling, and investigating disease mechanisms. However, iPSCs carrying specific mutations may limit their clinical applications due to certain inherent characteristics.
OBJECTIVE: To investigate the impact of MERTK mutations on hiPSCs and determine whether hiPSC-derived extracellular vesicles (EVs) influence anomalous cell junction and differentiation potential.
METHODS: We employed a non-integrating reprogramming technique to generate peripheral blood-derived hiPSCs with and hiPSCs without a MERTK mutation. Chromosomal karyotype analysis, flow cytometry, and immunofluorescent staining were utilized for hiPSC identification. Transcriptomics and proteomics were employed to elucidate the expression patterns associated with cell junction abnormalities and cellular differentiation potential. Additionally, EVs were isolated from the supernatant, and their RNA and protein cargos were examined to investigate the involvement of hiPSC-derived EVs in stem cell junction and differentiation.
RESULTS: The generated hiPSCs, both with and without a MERTK mutation, exhibited normal karyotype and expressed pluripotency markers; however, hiPSCs with a MERTK mutation demonstrated anomalous adhesion capability and differentiation potential, as confirmed by transcriptomic and proteomic profiling. Furthermore, hiPSC-derived EVs were involved in various biological processes, including cell junction and differentiation.
CONCLUSIONS: HiPSCs with a MERTK mutation displayed altered junction characteristics and aberrant differentiation potential. Furthermore, hiPSC-derived EVs played a regulatory role in various biological processes, including cell junction and differentiation.