Regenerating the Heart with Engineered Human Cardiac Tissue

描述(申请人提供)：心肌梗死后的心力衰竭主要是由于心肌细胞的死亡，这表明成功的基于细胞的治疗将取代心肌细胞来恢复心脏功能。为此，这项建议的重点是利用人类诱导多能干细胞(HiPSC)来源的心肌细胞在工程化心肌组织中生成大型人类心脏移植物，并提供治疗性生长因子和小分子药物的生物材料。我们用HiPSC-心肌细胞及其分泌的细胞外基质开发了无支架的工程化心脏组织。这些心脏组织“贴片”被植入梗死心脏的心外膜表面，但与宿主的结合微乎其微，心功能也没有改善。因此，这项建议旨在改善移植物的宿主血管化(目标1)、HiPSC来源的心肌细胞的存活和增殖(目标2)以及HiPSC-心肌细胞的力量产生(目标3)。在目标1中，我们将开发负载血管生成蛋白血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(BFGF)的可生物降解藻酸盐微球。这些材料将在体外和体内进行优化，以控制蛋白质的释放，并将其整合到心脏贴片中。 用于植入梗塞的大鼠心脏。在目标2中，我们将使用负载胰岛素样生长因子-1(IGF-1)、中性调节蛋白-1(NRG-1)和Y27632(Rho相关激酶的小分子抑制剂)的海藻酸盐微球来改善心脏补片形成期间和植入后HiPSC-心肌细胞的存活和增殖。在目标3中，我们使用化学、电和机械条件处理来促进HiPSC-心肌细胞肥大和收缩强度。目前的建议旨在解决基于细胞的心脏治疗的不足，并在方法上是创新的，使用可降解的生物材料在工程心脏组织中传递治疗性蛋白质/药物。这些迫在眉睫的研究目标将鼓励我作为一名独立调查员的发展。在指导的K99阶段，我将学习制造装载蛋白质和药物的藻酸盐微球。将控释系统与心脏组织工程相结合，将为我的研究生涯带来一个独特的利基市场。它是 我的长期职业目标是建立一个跨学科的心血管生物工程实验室，利用组织工程、生物材料、生理学、生物物理学和干细胞生物学方面的新技术来解决生物和医学问题。

DESCRIPTION (provided by applicant): Heart failure after a myocardial infarction is primarily due to death of cardiomyocytes, suggesting that successful cell-based therapies will replace cardiomyocytes to restore heart function. To that end, this proposal focuses on generating large human cardiac grafts using human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes in engineered cardiac tissue with biomaterial delivery of therapeutic growth factors and small molecule drugs. We have developed scaffold-free engineered cardiac tissue with hiPSC- cardiomyocytes and the extracellular matrix that they secrete. These cardiac tissue "patches" are implanted on the epicardial surface of infarcted hearts, but integration with the host is minimal and improved cardiac function is absent. Therefore, this proposal aims to improve host vascularization of the graft (Aim 1), survival and proliferation of hiPSC-derived cardiomyocytes (Aim 2), and force generation by hiPSC-cardiomyocytes (Aim 3). In Aim 1, we will develop biodegradable alginate microspheres loaded with the angiogenic proteins vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). These will be optimized in vitro and in vivo for controlled protein release and incorporated into cardiac patches for implantation in the infarcted rat heart. In Aim 2, we will use alginate microspheres loaded with insulin-like growth factor-1 (IGF-1), neuregulin-1 (NRG-1), and Y27632 (a small molecule inhibitor of Rho-associated kinase) to improve hiPSC- cardiomyocyte survival and proliferation during cardiac patch formation and after implantation. In Aim 3, we use chemical, electrical, and mechanical conditioning to promote hiPSC-cardiomyocyte hypertrophy and contractile strength. The current proposal aims to address deficiencies in cell-based cardiac therapy and is innovative in its approach, using degradable biomaterials for therapeutic protein/drug delivery within engineered cardiac tissue. These immediate research goals will encourage my development as an independent investigator. During the mentored K99 phase, I will learn to fabricate alginate microspheres loaded with proteins and drugs. Integrating controlled-release systems with cardiac tissue engineering will result in a unique niche for my research career. It is my long-term career goal to establish an interdisciplinary cardiovascular bioengineering lab that approaches biological and medical problems with novel technologies in tissue engineering, biomaterials, physiology, biophysics, and stem cell biology.