Human iPSC-derived endothelial cells as Vascular Therapeutics

项目摘要 外周动脉疾病(PAD)在55年内困扰着大约15%的美国人口 年纪大了。最初以间歇性跛行为特征，渐进性垫导致无情 休息疼痛和溃疡，这种情况被称为严重肢体缺血(CLI)。尽管取得了进步 在经皮介入或外科治疗中，多达一半的CLI患者被截肢 一年之内。病理生理学上，通过直接血管形成减轻缺血 将是最理想的治疗方法。虽然使用成体干细胞或祖细胞进行细胞治疗一直是 作为PAD的一种新的血管重建疗法的研究，临床试验表明没有或很少 效果。与此同时，人类诱导多能干细胞(HiPSCs)被发现并显示 几乎与人类胚胎干细胞(HESCs)的特性相似。因为真的 针对靶细胞的分化能力，HiPSCs已成为一种很有前途的治疗选择 用于细胞治疗。我们和其他人开发了一个协议来生成非整合的HiPSCs 来自少量外周血液的上体载体。我们进一步开发了一个系统来 以临床相容的方式将HiPSC分化为内皮细胞(HiPSC-ECs) 在动物模型上展示了它们的长期血管形成作用和治疗作用。 该项目的最终目标是开发HiPSC-ECs作为治疗药物。 治疗严重的外周动脉疾病。具体地说，在这个提案中，我们将生成HiPSC 从PAD患者和正常志愿者外周血中诱导分化为HiPSC-ECs 使用我们既定的程序。然后我们将确定这些HiPSC的身份和效力- ECS使用体外和体内测试，特别是关注是否有类似的效力 患者来源的和正常志愿者来源的HiPSC-ECs之间的差异。如果表现出相似之处，更安全的 将为这种疾病开发自体方法。 尽管HiPSC-ECs的治疗潜力令人难以置信，但它的细胞疗法尚未发表 或迄今为止在全球范围内提出的建议。我们的调查团队拥有所有必要的专业知识和 将我们的发现转移到未来的临床试验中。这笔赠款将在以下方面发挥作用 从长凳到床边的HiPSCs，我们相信这将是最重要的细胞来源 在不久的将来进行治疗。此外，这项工作也是HiPSC治疗应用的第一步。 欧共体。有许多潜在的疾病需要血管供应，如缺血性心脏 疾病、中风、糖尿病视网膜病变、糖尿病神经病变和伤口愈合，这些都将是所有的 从这一发展中受益的候选人。因此，这项研究的影响将是巨大的。

Project Summary Peripheral arterial disease (PAD) afflicts approximately 15% of the U.S. population over 55 years of age. Initially characterized by intermittent claudication, progressive PAD results in unrelenting rest pain and ulceration, a condition referred to as critical limb ischemia (CLI). Despite advances in percutaneous intervention or surgical treatment, up to half of these CLI patients are amputated within one year. Pathophysiologically, alleviation of ischemia through direct vessel formation would be an optimal treatment. While cell therapy with adult stem or progenitor cells has been investigated as a new revascularization therapy for PAD, clinical trials showed no or minimal effects. Meanwhile, human induced pluripotent stem cells (hiPSCs) were discovered and showed almost similar properties to human embryonic stem cells (hESCs). Because of genuine differentiation capacity to target cells, hiPSCs have emerged as a promising therapeutic option for cell therapy. We and others developed a protocol to generate hiPSCs with non-integrating episomal vectors from a small amount of peripheral blood. We further developed a system to differentiate hiPSCs into endothelial cells (hiPSC-ECs) in a clinically compatible manner and demonstrated their long-term vessel-forming effects and therapeutic effects on animal models. The ultimate goal of this project is to develop hiPSC-ECs as a therapeutic agent for treating severe peripheral artery disease. Specifically, in this proposal, we will generate hiPSCs from peripheral blood of PAD patients and normal volunteers, differentiate them into hiPSC-ECs using our established procedure. We will then determine the identity and potency of these hiPSC- ECs using both in vitro and in vivo assays, particularly focusing on whether there is similar potency between the patient-derived and normal-volunteer-derived hiPSC-ECs. If shown similarity, a safer autologous approach will be developed for this disease. Despite its incredible therapeutic potential, cell therapy with hiPSC-ECs was not published or proposed worldwide to date. Our team of investigators have all the necessary expertise and experience to move our discovery into a future clinical trial. This grant will bridge the utility of hiPSCs from bench to bedside, which, we believe, will be the most significant source of cell therapy in the near future. Moreover, this work is a first step for therapeutic application of hiPSC- EC. There are many potential diseases that need a vascular supply such as ischemic heart disease, stroke, diabetic retinopathy, diabetic neuropathy, and wound healing, which will be all candidates that benefit from this development. Thus, the impact of this study will be huge.