Effect of Microgravity on Drug Responses Using Engineered Heart Tissues

项目总结 组织工程化器官或功能组织样合体对我们理解 允许细胞在三维(3-D)中迁移、发育和成熟的细胞壁龛。传统的两个- 二维(2-D)哺乳动物细胞培养并不代表形成 是正常细胞功能的基础。3-D环境促进各向同性的细胞间通信，提供 细胞外引导，从结构基质支架，并允许时空重构。我们的特定 人们感兴趣的是使用工程心脏研究微重力对心脏功能的影响 组织(EHTS)。由于这些组织工程平台自下而上支持多细胞体系结构 UP的方法，从原始状态理解心脏发育的机制是至关重要的。 虽然动物模型被广泛用于研究对治疗的生物学反应，但固有的 人类和动物生物学上的差异，以及动物不太可能发展成 人类疾病限制了验证研究结果的能力。人诱导多能干细胞 已经成为将细胞从胚胎状态驱动到任何体细胞类型的不可或缺的工具。我们的 实验室在培养HiPSC来源的心肌细胞(HiPSC-CMS)和建模方面的重点和专业知识 心肌病使人们对心力衰竭的几种罕见和常见原因有了更深入的了解。至 维持组织特有的微环境，分离的细胞必须在与生理相关的 三维细胞外基质(ECM)。在第一阶段(UG3)，我们将从健康患者中产生HiPSC-CMS 属于不同的种族群体(高加索人、西班牙人和非裔美国人)。HiPSC-CMS将是 用于制造我们特性良好的EHT平台，以了解影响心脏的细胞机制 在微重力和地球重力下都能发挥作用。心脏衰弱引起的心功能改变 暴露在微重力下的样品中的肌肉将与分子和电生理相匹配 观察到的缺血性心肌病的疾病类型。在第二阶段(UH3)，特征良好的 微重力引起的疾病表型将被转化到心脏组织阵列(HTA)上进行筛查 以高通量的方式获得潜在的候选药物。这项拟议的研究将首次揭示关键 功能和分子差异驱动心脏组织在EHT组件上的表型变化 微重力的影响。

PROJECT SUMMARY Tissue engineered organs or functional tissue-like ensembles contribute significantly to our understanding of cellular niches that allow cells to migrate, develop and mature in three dimensions (3-D). Conventional two- dimensional (2-D) mammalian cell culture does not represent the physiological environments that form the basis for normal cell function. A 3-D environment promotes isotropic cell-cell communications, provides extracellular guidance from structural matrix scaffolding, and allows spatiotemporal remodelling. Our specific interest is in investigating the effects of microgravity on heart function with the use of Engineered Heart Tissues (EHTs). Since these tissue engineering platforms support multicellular architecture from a ‘bottom- up’ approach, it is critical to understand the mechanisms of heart development from a primordial state. Although animal models are used widely to investigate biological responses to therapeutics, inherent differences between human and animal biology combined with the unlikelihood of animals developing a human disease limit the ability to validate research findings. Human induced pluripotent stem cells (hiPSCs) have emerged as an indispensable tool to drive cells from an embryonic state to any somatic cell type. Our laboratory’s focus and expertise in generating hiPSC-derived cardiomyocytes (hiPSC-CMs) and modelling of cardiomyopathies has yielded deeper insight into several rare and common causes of heart failure. To maintain a tissue-specific microenvironment, dissociated cells must be cultured in a physiologically relevant 3-D extracellular matrix (ECM). In the first phase (UG3), we will generate hiPSC-CMs from healthy patients belonging to diverse racial groups (Caucasians, Hispanics, and African Americans). The hiPSC-CMs will be used to fabricate our well-characterized EHT platforms, to understand cellular mechanisms that affect cardiac function both under microgravity and earth’s gravity. Alterations in cardiac function due to weakened heart muscles in the samples exposed to microgravity will be matched with molecular and electrophysiological disease patterns observed in ischemic cardiomyopathy. In the second phase (UH3), the well-characterized microgravity-induced disease phenotype will be translated on Heart Tissue Arrays (HTA) to screen for potential drug candidates in a high-throughput manner. The proposed study will for the first time reveal key functional and molecular differences that drive phenotypic changes in heart tissues on EHT assemblies under influence of microgravity.