ECM Shrink Wrapped Human Cardiomyocytes and Endothelial Cells to Accelerate Myocardial Regeneration

心肌梗死（MI）是美国心脏相关死亡的主要原因， 急性事件具有心律失常、中风和充血性心力衰竭慢性风险。修复心脏是 因为心肌细胞是有丝分裂后的，不能增殖以再生受损的组织。 最近的研究表明，人心肌细胞可以来源于胚胎干细胞（ES）， 诱导多能干细胞（iPS），以及从其他细胞转分化。然而，生存和 将这些心肌细胞功能性整合到典型的血管化心肌中仍然是一个主要的， 未解决的挑战该提案描述了一种突破性的治疗性细胞递送， 纳米结构的细胞外基质（ECM）支架中的每个细胞都是为了提高存活率而定制的， 肌生成和整合到梗死心肌中。我们方法的关键创新在于能够 设计50-100 nm厚ECM片，其具有确定的蛋白质组成和形状，并将其包裹在 单个细胞或小细胞系。这是对当前封装技术的改进 因为我们可以在心肌细胞和内皮细胞周围构建ECM， 在健康的心脏中包围着这些细胞。这是至关重要的，因为ECM是整合素的主要调节器 结合、生长因子螯合和机械转导。我们的初步结果表明， 一种新的表面引发的组装技术可以从一系列基质蛋白质构建ECM纳米支架 收缩包裹心肌细胞和内皮细胞。此外，我们已经证明使用角膜内皮细胞 我们可以在体内有效地输送细胞。本提案将在这些成果的基础上，实现三个目标： 主要目标。第一，发展ECM纳米支架技术，收缩包裹心肌细胞和内皮细胞， 纤维连接蛋白、层粘连蛋白和IV型胶原蛋白的工程化层中的细胞，其与天然细胞中的基质相匹配， 心肌第二，询问ECM纳米支架的组成、大小和细胞群体对细胞增殖的作用。 最大化3D组织中的肌肉形成、预血管化和收缩性。展望未来，实现 这些目标将导致可注射细胞递送技术，其具有增强的促进细胞增殖的能力。 保留、存活、整合、肌生成和血管化的心肌细胞和内皮细胞进入 受伤的心这将产生深远的影响，导致临床相关的治疗 降低MI和心血管疾病患者发病率和死亡率的策略

Myocardial infarction (MI) is a major cause of cardiac-related death in the US and those fortunate to survive the acute event suffer from chronic risk of arrhythmia, stroke and congestive heart failure. Repairing the heart is difficult because cardiomyocytes are post-mitotic and cannot proliferate in order to regenerate damaged tissue. Recent work has demonstrated that human cardiomyocytes can be derived from embryonic stem (ES) and induced pluripotent stem (iPS) cells, as well as transdifferentiated from other cells. However, survival and functional integration of these cardiomyocytes into stereotypical vascularized myocardium is still a major and unresolved challenge. This proposal describes a breakthrough towards therapeutic cell delivery by wrapping each cell in a nanostructured extracellular matrix (ECM) scaffold tailor made for enhancing survival, myogenesis and integration into infarcted myocardium. The key innovation in our approach is the ability to engineer 50-100 nm thick sheets of ECM with defined protein composition and shape and wrap this around individual cells or small cell ensembles. This is an improvement over current encapsulation technology because we can build the ECM around cardiomyocytes and endothelial cells to mimic the ECM that naturally surrounds these cells in the healthy heart. This is critical, because the ECM is a primary regulator of integrin binding, growth factor sequestration and mechanotransduction. Our preliminary results demonstrate that our novel surface-initiated assembly technique can build an ECM nano-scaffold from a range of matrix proteins and shrink wrap cardiomyocytes and endothelials cells. Further, we have shown using corneal endothelial cells that we can effectively delivery cells in vivo. This proposal will build upon these results by achieving three primary aims. One, develop the ECM nano-scaffold technology to shrink wrap cardiomyocytes and endothelial cells in engineered layers of fibronectin, laminin and collagen type IV that match the matrix in the native myocardium. Two, interrogate the role of ECM nano-scaffold composition, size and cell population on maximizing muscle formation, pre-vascularization and contractility in 3D tissue. Looking forward, achieving these aims will result in an injectable cell delivery technology that has enhanced capability to promote the retention, survival, integration, myogenesis and vascularization of cardiomyocytes and endothelial cells into the injured heart. This would have profound consequences by leading towards clinically-relevant therapeutic strategies to decrease morbidity and mortality in MI and cardiovascular disease patients