Graduate Institute of Biomedical Materials & Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, 301 Yuantong Road, Taipei, 235603, Taiwan.
Department of Surgery, School of Medicine, College of Medicine, Taipei Medical University, 250 Wuxing Street, Taipei, 110, Taiwan.
Stem Cell Res Ther. 2024 Oct 12;15(1):363. doi: 10.1186/s13287-024-03983-y.
Cell therapy can protect cardiomyocytes from hypoxia, primarily via paracrine secretions, including extracellular vesicles (EVs). Since EVs fulfil specific biological functions based on their cellular origin, we hypothesised that EVs from human cardiac stromal cells (CMSCLCs) obtained from coronary artery bypass surgery may have cardioprotective properties.
This study characterises CMSCLC EVs (C_EVs), miRNA cargo, cardioprotective efficacy and transcriptomic modulation of hypoxic human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). C_EVs are compared to bone marrow mesenchymal stromal cell EVs (B_EVs) which are a known therapeutic EV type.
Cells were characterised for surface markers, gene expression and differentiation potential. EVs were compared for yield, phenotype, and ability to protect hiPSC-CMs from hypoxia/reoxygenation injury. EV dose was normalised by both protein concentration and particle count, allowing direct comparison. C_EV and B_EV miRNA cargo was profiled and RNA-seq was performed on EV-treated hypoxic hiPSC-CMs, then data were integrated by multi-omics. Confirmatory experiments were carried out using miRNA mimics.
At the same dose, C_EVs were more effective than B_EVs at protecting CM integrity, reducing apoptotic markers, and cell death during hypoxia. While C_EVs and B_EVs shared 70-77% similarity in miRNA content, C_EVs contained unique miRNAs, including miR-202-5p, miR-451a and miR-142-3p. Delivering miRNA mimics confirmed that miR-1260a and miR-202/451a/142 were cardioprotective, and the latter upregulated protective pathways similar to whole C_EVs.
This study demonstrates the potential of cardiac tissues, routinely discarded following surgery, as a valuable source of EVs for myocardial infarction therapy. We also identify miR-1260a as protective of CM hypoxia.
细胞疗法可以通过旁分泌等方式保护心肌细胞免受缺氧的影响，其中包括细胞外囊泡（EVs）。由于 EVs 基于其细胞来源而具有特定的生物学功能，因此我们假设，从冠状动脉旁路手术中获得的人心肌基质细胞（CMSCLCs）的 EVs 可能具有心脏保护作用。
本研究旨在描述 CMSCLC EVs（C_EVs）、miRNA cargos、缺氧状态下人诱导多能干细胞衍生心肌细胞（hiPSC-CMs）的心脏保护功效和转录组调节。并将 C_EVs 与骨髓间充质基质细胞 EVs（B_EVs）进行比较，B_EVs 是一种已知的治疗性 EV 类型。
对细胞进行表面标志物、基因表达和分化潜能的鉴定。比较 EV 的产量、表型和保护 hiPSC-CMs 免受缺氧/复氧损伤的能力。通过蛋白浓度和颗粒计数对 EV 进行归一化，以实现直接比较。对 C_EV 和 B_EV 的 miRNA cargos 进行分析，并对 EV 处理后的缺氧 hiPSC-CMs 进行 RNA-seq 分析，然后通过多组学进行数据整合。使用 miRNA 模拟物进行验证实验。
在相同剂量下，C_EVs 比 B_EVs 更能有效保护 CM 完整性，减少缺氧时的凋亡标志物和细胞死亡。虽然 C_EVs 和 B_EVs 的 miRNA 含量有 70-77%的相似性，但 C_EVs 含有独特的 miRNA，包括 miR-202-5p、miR-451a 和 miR-142-3p。递送 miRNA 模拟物证实，miR-1260a 和 miR-202/451a/142 具有心脏保护作用，后两者上调了与整个 C_EVs 相似的保护途径。
本研究证明了心脏组织作为心肌梗死治疗中 EV 有价值来源的潜力，这些组织通常在手术后被丢弃。我们还发现 miR-1260a 对 CM 缺氧具有保护作用。
