Microfluidic single-cell analysis shows that porcine induced pluripotent stem cell-derived endothelial cells improve myocardial function by paracrine activation.
Microfluidic single-cell analysis shows that porcine induced pluripotent stem cell-derived endothelial cells improve myocardial function by paracrine activation.
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DOI：
10.1161/circresaha.112.269001
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发表时间：
2012-09-14
影响因子：
20.1
通讯作者：
Wu JC
中科院分区：
文献类型：
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作者：
Gu M;Nguyen PK;Lee AS;Xu D;Hu S;Plews JR;Han L;Huber BC;Lee WH;Gong Y;de Almeida PE;Lyons J;Ikeno F;Pacharinsak C;Connolly AJ;Gambhir SS;Robbins RC;Longaker MT;Wu JC
诱导多能干细胞(IPSCs)为开发针对患者的心血管疾病治疗方法提供了巨大的希望。然而，临床翻译将需要对大型动物IPSC模型进行临床前优化和验证。成功地从猪IPSCs中分离出内皮细胞，并证明其在治疗心肌缺血方面的潜在用途。猪脂肪基质细胞被重新编程以产生猪IPSCs(PiPSCs)。免疫组织化学、定量聚合酶链式反应、微阵列杂交和血管生成实验证实，PIPSC来源的内皮细胞(PIPSC-ECs)具有与自体猪主动脉内皮细胞相似的形态和功能特性。为了证明其治疗潜力，将PIPSC-ECs移植到心肌梗死(MI)小鼠体内。与对照组相比，移植了PIPSC-ECs的动物在超声心动图(4周时缩短率：27.2±1.3%比22.3±1.1%；P<；0.001)和磁共振成像(4周时射血分数：45.8±1.3%比42.3±0.9%；P<；0.05)方面有了显著的改善。蛋白质定量分析和微流控单细胞PCR图谱显示，PIPSC-ECs在缺血微环境中释放促血管生成因子和抗凋亡因子，分别促进新生血管形成和心肌细胞存活。旁分泌因子的释放在不同的移植细胞亚群之间有显著差异，这表明特定细胞群体的移植可能会导致更大的功能恢复。总之，这是第一项成功区分PiPSCs和PiPSCs的研究，并证明PIPSC-ECs移植通过旁分泌激活改善了MI后的心功能。这些大型动物IPSC模型的进一步发展将对其治疗潜力产生重要的见解，并加速基于自体IPSC的治疗的临床转化。
Induced pluripotent stem cells (iPSCs) hold great promise for the development of patient-specific therapies for cardiovascular disease. However, clinical translation will require preclinical optimization and validation of large animal iPSC models. To successfully derive endothelial cells from porcine iPSCs and demonstrate their potential utility for the treatment of myocardial ischemia. Porcine adipose stromal cells were reprogrammed to generate porcine iPSCs (piPSCs). Immunohistochemistry, quantitative PCR, microarray hybridization, and angiogenic assays confirmed that piPSC-derived endothelial cells (piPSC-ECs) shared similar morphological and functional properties as endothelial cells isolated from the autologous pig aorta. To demonstrate their therapeutic potential, piPSC-ECs were transplanted into mice with myocardial infarction (MI). Compared to control, animals transplanted with piPSC-ECs showed significant functional improvement measured by echocardiography (fractional shortening at week 4: 27.2±1.3% vs. 22.3±1.1%; P<0.001) and magnetic resonance imaging (ejection fraction at week 4: 45.8±1.3% vs. 42.3±0.9%; P<0.05). Quantitative protein assays and microfluidic single cell PCR profiling showed that piPSC-ECs released pro-angiogenic and anti-apoptotic factors in the ischemic microenvironment, which promoted neovascularization and cardiomyocyte survival, respectively. Release of paracrine factors varied significantly among subpopulations of transplanted cells, suggesting that transplantation of specific cell populations may result in greater functional recovery. In summary, this is the first study to successfully differentiate piPSCs-ECs from piPSCs and demonstrate that transplantation of piPSC-ECs improved cardiac function following MI via paracrine activation. Further development of these large animal iPSC models will yield significant insights into their therapeutic potential and accelerate the clinical translation of autologous iPSC-based therapy.