Vascular Regeneration with Human Pluripotent Stem Cell-derived Vascular Cells and Engineering Approaches

项目摘要 缺血性心血管疾病是美国发病率和死亡率的主要原因。尽管 尽管在治疗学方面取得了进步，但治疗具有严重病症的患者仍然远远不是最佳的。最近，Cell 对于那些不能进行介入或手术治疗的晚期病例， 治疗能够有效地使缺血区域再血管化。 人多能干细胞（hPSC），其包括人胚胎干细胞（hESC）和人胚胎干细胞（hPSC）。 诱导多能干细胞（hiPSC）已经成为血管再生的有希望的候选者， 它们具有强的靶细胞分化能力以及旁分泌效应。因此，调查人员 开发了各种方案以将hPSC分化为内皮细胞（EC）谱系细胞。我们已经开发 一种完全确定的、不含异种成分的细胞培养系统， 细胞（ECs）产量高。我们进一步证明了这些hPSC衍生的EC（hPSC-EC）具有鲁棒性和特异性。 延长体内血管形成活性。然而，这种方法的一个警告是， 主要局限于毛细血管水平，没有周细胞。为了实现最佳血管化，更稳定、更大 船只也是必要的。在以前的细胞治疗研究中，这方面几乎没有得到解决。因此，我们认为， 我们最近也通过使用确定的培养系统产生了人PSC衍生的SMCs（hPSC-SMCs 并观察它们作为血管周细胞和SMC对血管形成的贡献。 细胞治疗的另一个重要障碍是移植细胞的短期存活。克服 这个问题，我们和其他人已经研究了生物工程细胞疗法，并证明了它的有效性， 细胞存活和功能。然而，注射细胞的不均匀和局部分布出现了另一个问题， 问题.最近，我们开发了一种新型的可生物降解的杂化共聚物， 癸二酸甘油酯（PGS），其进一步与藻酸盐制成微珠形式。我们把这一共同- 聚合物作为AlGPM。这种混合聚合物是可生物降解的，并消除最小的炎症反应。此外，委员会认为， 其微珠形式促进了包封细胞在体内的广泛和均匀分布。 因此，在这项研究中，我们将解决目前缺血性脑血管病细胞治疗的两个未满足的需求， 血管疾病首先，我们将使用hPSC-EC和hPSC-SMC两者来诱导不仅裸细胞的形成， 毛细血管，而且周细胞覆盖的毛细血管和SMC覆盖的小动脉。第二，我们将开发新的 生物材料可以增强细胞在体内的存活和分布，以最大化稳定的血管形成， 治疗效果具体来说，我们将研究这两种细胞类型的组合是否与 AlGPM水凝胶微珠能够对血管再生发挥最佳效果。长期 本研究的目的是利用hPSC衍生的血管细胞开发临床适用的再生疗法 结合生物工程技术。

Project Summary Ischemic cardiovascular diseases are the leading causes of morbidity and mortality in the USA. Despite advancement in therapeutics, treating patients with severe conditions are still far from optimal. Recently, cell therapy emerged as a promising option for those advanced cases for which no interventional or surgical therapy is able to effectively revascularize the ischemic areas. Human pluripotent stem cells (hPSCs), which include human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), have emerged as a promising candidate for vascular regeneration as they have strong target cell differentiation capacity as well as paracrine effects. Thus, investigators have developed various protocols to differentiate hPSCs into endothelial cell (EC)-lineage cells. We have developed a fully defined, xenogeneic ingredient-free cell culture system that can generate purified functional endothelial cells (ECs) at high yield. We further demonstrated that these hPSC-derived ECs (hPSC-ECs) have robust and prolonged vessel-forming activities in vivo. However, one of the caveats of this approach is that their contribution is mainly restricted to the capillary level without pericytes. For optimal vascularization, more stable and larger vessels are also necessary. In previous cell therapy studies, this aspect was virtually unaddressed. Therefore, we recently generated human PSC-derived SMCs (hPSC-SMCs) by using a defined culture system as well and observed their contribution to vessel formation as vascular pericytes and SMCs. Another important barrier for cell therapy is short-term survival of the transplanted cells. To overcome this problem, we and others have investigated bioengineered cell therapy and demonstrated its effectiveness for cell survival and function. However, uneven and localized distribution of the injected cells emerged as another problem. Recently, we have developed a novel biodegradable hybrid copolymer consisting of gelatin and poly glycerol sebacate (PGS), which was further made into a microbead form with alginate. We refer to this co- polymer as AlGPM. This hybrid polymer is biodegradable and elicits minimal inflammatory responses. Moreover, its microbead form promotes wide and homogeneous distribution of encapsulated cells in vivo. Accordingly, in this study, we will address two unmet needs of the current cell therapy for ischemic vascular disease. First, we will use both hPSC-ECs and hPSC-SMCs to induce formation of not only bare capillaries but also pericyte-covered capillaries and SMC-covered arterioles. Second, we will develop a new biomaterial that can enhance cell survival and distribution in vivo to maximize stable vessel formation and therapeutic effects. Specifically, we will investigate whether a combination of these two cell types with AlGPM hydrogel microbeads is able to exert the optimal effects on vascular regeneration. The long-term goal of this study is to develop clinically applicable regenerative therapy using hPSC-derived vascular cells combined with bioengineering technologies.