Cardiac Regeneration with Bioengineered Human Stem Cells

 描述(申请人提供)：人类多能干细胞(HPSCs)，包括胚胎干细胞(HESCs)和诱导多能干细胞(HiPSCs)，为心脏修复提供了前所未有的前景；然而，在临床应用之前，必须克服许多障碍。目前将心肌细胞(CMS)从hPSCs中分化出来的方法会导致细胞群体的异质性，包括未分化或非CM细胞，这可能会带来肿瘤或异常组织形成的风险。对于治疗用途，心室CMS(VCM)是理想的，因为VCM是心肌梗死中受影响最广泛的细胞类型，也是产生心脏收缩力量的主要来源。虽然已经尝试了各种策略来进行CM浓缩，但这些策略在纯度或特异性方面存在缺陷。我们最近开发了一种新的技术，通过使用分子信标(MBS)靶向CM细胞内的mRNAs来纯化CMS，这将使我们能够分离出所需的CMS用于治疗。细胞治疗的另一个障碍是移植细胞在受损心肌中的低保留率。为了克服这一问题，人们尝试将CMS与生物材料或与其他细胞共移植。我和我的合作者一直致力于开发以两亲多肽(PA)为基础的纳米结构生物材料来设计心血管细胞的技术，以增强细胞治疗的效果。该方案的目的是通过两种新的方法开发一种利用hPSC来源的VCM进行心脏修复的策略：用MBS纯化hPSC来源的VCM，并用自组装的纳米基质凝胶和其他细胞(包括hPSC来源的ECs和成纤维细胞)来设计这些CMS。在目标1中，我们试图通过使用特定的MBS来区分hPSC来纯化VCM。在目标2中，我们将利用模拟细胞外基质的自组装纳米基质凝胶来工程纯化的hPSC来源的CMS、hPSC来源的ECs和其他支持性细胞，并确定最佳的细胞-纳米基质构建，其中包含生长因子以增强细胞活力、血管形成和细胞成熟。在目标3中，我们将确定hPSC来源的工程化心血管细胞构建物对受损心肌的治疗作用，并阐明其潜在的机制。我们预计，拟议的实验结果将为新型干细胞疗法在心脏修复中的作用提供新的见解。

 DESCRIPTION (provided by applicant): Human pluripotent stem cells (hPSCs), which include embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), offer unprecedented promise for cardiac repair; however, many hurdles must be overcome before clinical application is possible. Current methods for differentiating cardiomyocytes (CMs) from hPSCs result in heterogeneous populations of cells, including undifferentiated or non-CM cells, which may pose a risk for tumor or aberrant tissue formation. For therapeutic use, ventricular CMs (vCMs) are ideal because vCMs are the most extensively affected cell type in myocardial infarction and the major source for generating cardiac contractile forces. Although various strategies have been attempted for CM enrichment, these strategies have drawbacks in terms of purity or specificity. We recently developed a novel technology to purify CMs by targeting CM-specific intracellular mRNAs using molecular beacons (MBs), which will enable isolation of the desired CMs for use in therapy. Another obstacle to cell therapy is low retention of transplanted cells in injured myocardium. To overcome this problem, engineering of CMs with biomaterials or co-transplantation with other cells has been attempted. My co-investigator and I have been working to develop technologies for engineering cardiovascular cells with peptide amphiphile (PA)-based nanostructured biomaterials for enhancing the effects of cell therapy. The goal of this proposal is to develop a strategy for cardiac repair with hPSC-derived vCMs by two novel approaches: to purify hPSC-derived vCMs with MBs and to engineer these CMs with self-assembled nanomatrix gels and other cells including hPSC-derived ECs and fibroblasts. In aim 1, we seek to purify vCMs from differentiating hPSCs using specific MBs. In aim 2, we will engineer purified hPSC-derived CMs, hPSC-derived ECs and other supportive cells with extracellular matrix-mimicking self-assembled nanomatrix gel which contains growth factors to enhance cell viability, vascularization, and cell maturation, and determine the optimal cell- nanomatrix constructs. In aim 3, we will determine the therapeutic effects of engineered hPSC- derived cardiovascular cell constructs on injured myocardium and elucidate the underlying mechanisms. We anticipate that the results of the proposed experiments will yield new insight into the role of novel stem cell therapy for cardiac repair.