Regenerating the Heart with Engineered Human Cardiac Tissue

项目总结/摘要 心肌梗死后心力衰竭主要是由于心肌细胞死亡，这表明， 成功的基于细胞的疗法将取代心肌细胞以恢复心脏功能。为此，这 一项提案的重点是使用人类诱导多能干细胞产生大型人类心脏移植物 具有治疗性生长的生物材料递送的工程化心脏组织中的（hiPSC）衍生的心肌细胞 因子和小分子药物。我们用hiPSC开发了无支架的工程化心脏组织- 心肌细胞和它们分泌的细胞外基质。这些心脏组织“补丁”被植入 梗死心脏的心外膜表面，但与宿主的整合是最小的，并改善心脏功能 缺席。因此，该提议旨在改善移植物的宿主血管化（目标1）、存活率和存活率。 hiPSC衍生的心肌细胞的增殖（Aim 2）和hiPSC-心肌细胞的力产生（Aim 3）。 目的一：研制可生物降解的海藻酸钠微球， 内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）。这些将在体外进行优化 并在体内用于控制蛋白质释放，并掺入心脏补片中用于植入梗塞的 鼠心在目标2中，我们将使用负载胰岛素样生长因子-1（IGF-1）的藻酸盐微球， 神经调节蛋白-1（NRG-1）和Y27632（Rho相关激酶的小分子抑制剂），以改善hiPSC-1表达。 在心脏补片形成期间和植入后的心肌细胞存活和增殖。在目标3中， 使用化学、电和机械调节来促进hiPSC-心肌细胞肥大， 收缩力目前的提案旨在解决基于细胞的心脏治疗的缺陷， 创新的方法，使用可降解的生物材料用于治疗性蛋白质/药物递送， 工程心脏组织这些直接的研究目标将鼓励我作为一个 独立调查员在辅导K99阶段，我将学习制作海藻酸盐微球 装载着蛋白质和药物将控释系统与心脏组织工程相结合， 为我的研究事业创造了一个独特的利基市场。建立一个跨学科的， 心血管生物工程实验室，采用新技术解决生物和医学问题， 组织工程学、生物材料学、生理学、生物物理学和干细胞生物学。

Project Summary/Abstract 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.