Morphogenetic Self-Assembly of Human Heart Organoids

项目总结 出于实验目的无法获得人类胚胎是合理的，这是一个长期存在的障碍。 对人体器官发生的体外研究。为了绕过这一僵局，这款R21的总体目标是 建议建立一个强大的新的三维实验模型系统，它概括了关键的结构和 胚胎生态位环境的生物物理要素使人类心脏研究成为可能 形态发生和发育。我们提出了一个创新的生物反应器设计的协同组合， 最新的人类多能干细胞生物学和尖端的人类心脏组织工程 技术，以合理的生物工程原则为基础的量化方法。战略是发展 一种基于微流体的生物反应器系统，用于培养人心内膜管、心肌和心脏凝胶 多能干细胞(HPSC)来源的心肌细胞、成纤维细胞和心内膜细胞 体外心管形成的早期发育阶段。通过提供流体剪切力、机械 限制和形态梯度，与发育中的心管所经历的相似，并通过 在模仿原始体的起始位置用空间分隔的细胞类型播种生物反应器 解剖学，我们的目标是提供一个利基环境，在其中他们可以开始建造心脏，就像他们在 大自然。本提案的两个具体目标与R21探索性/开发性目标一致 生物工程研究资助机制(PA-16-040)：目标1是开发具有以下特性的微流控生物反应器 用于创造和评价工程形态发生人的边界约束和流量控制 心脏器官。目的2建立人先天性心管畸形的体外模型， 探索已知的化学和遗传干预导致体内的异质性。这种探索性的和 发展提案提供了亟需的、潜在的颠覆性飞跃 现有的体外人体模型系统的复杂性。研究团队的多学科专业知识， 与可用的最先进的资源和设施相结合，提供了一个独特的机会来克服 预期的技术挑战，并成功实现拟议的目标。因此，这被认为是 一项高风险、高影响的提案，可能会为理解心脏疾病的过程提供新的工具和见解 管形成，用于创造更逼真的多组织心脏器官，并最终用于研究疾病 与心脏结构缺陷相关的研究使用了一种独特的体外解剖三维人体心脏模型系统。

PROJECT SUMMARY A justifiable lack of access to human embryos for experimental purposes has presented a longstanding barrier to the in vitro investigation of human organogenesis. To circumvent this impasse, the overall goal of this R21 proposal is to establish a powerful new 3-D experimental model system that recapitulates key structural and biophysical elements of the embryonic niche environment to enable investigation of human cardiac morphogenesis and development. We propose a synergistic combination of innovative bioreactor design, state-of-the-art human pluripotent stem cell biology, and cutting-edge human cardiac tissue engineering technology, with a quantitative approach based on sound bioengineering principles. The strategy is to develop a microfluidics based bioreactor system to grow endocardial tubes, myocardium, and cardiac jelly using human pluripotent stem cell (hPSC) derived cardiomyocytes, fibroblasts and endocardial cells in order to study the early developmental stages of heart tube formation in vitro. By providing fluid shear forces, mechanical constraints, and morphogen gradients similar to those experienced by the developing heart tube, and by seeding the bioreactor with spatially compartmentalized cell types in starting positions that mimic the primitive anatomy, we aim to provide a niche environment in which they can begin building a heart as they would in nature. The two specific aims of this proposal are consistent with the R21 Exploratory/Developmental Bioengineering Research Grant mechanism (PA-16-040): Aim 1 is to develop a microfluidic bioreactor with boundary constraints and flow control for the creation and evaluation of engineered morphogenetic human heart organoids. Aim 2 is to establish an in vitro model of human congenital heart tube malformation, exploring chemical and genetic interventions known to cause heterotaxy in vivo. This exploratory and developmental proposal offers a desperately needed and potentially disruptive leap forward in the sophistication of available in vitro human model systems. The multidisciplinary expertise of the research team, combined with the available state-of-the-art resources and facilities, provides a unique opportunity to overcome the anticipated technical challenges and successfully achieve the proposed aims. This is therefore considered a high-risk, high-impact proposal, likely to yield new tools and insights for understanding the process of cardiac tube formation, for creating more realistic multi-tissue heart organoids, and for eventually studying diseases related to cardiac structural defects using a unique anatomical in vitro 3-D human heart model system.