Polymer Microarrays for Stem Cell Cardiac Differentiation

项目主任/首席调查员(最后、第一、中间)：Vyavahare Narendra R. 项目总结 目前无法有效地从人类胚胎中获得足够数量的成熟心肌细胞 干细胞(HESCs)和人诱导多能干细胞(HiPSCs)的应用受到严重限制。 人类干细胞技术用于治疗心血管疾病，这是全球主要的死亡原因。 已经进行了大量的研究来设计可溶性因子，如生长因子和小分子 分子，以诱导hESCs和hiPSCs的心脏分化。相比之下，几乎没有做过什么工作来 优化不溶性因素，例如细胞生长的底物，以促进心脏分化。 此外，目前来源于hESCs和hPSCs的心肌细胞在结构和功能上类似于 人胚胎/新生儿期心肌细胞(即未成熟心肌细胞)，临床应用有限 申请。因此，我们将追求两个具体目标：1)聚合物的高通量评估 促进hESCs心脏分化的底物，以及2)促进hESC的终末分化。 通过模拟生物化学和生物物理刺激的关键方面获得未成熟的心肌细胞 心脏发育。我们在目标1中假设，随着高通量筛选聚合物库 已知的促进hESC克隆生长的底物，能够促进心脏分化的底物 可以鉴定出人类胚胎干细胞。我们在目标2中假设我们可以促进hESC来源的未成熟细胞的成熟。 在发育中的心脏中模拟生化和生物物理刺激的心肌细胞。这项研究是 创新：我们将首次利用新兴的聚合物微阵列技术来开发明确的 以高通量方式进行底物培养，以促进人类胚胎干细胞的心脏分化。此外，我们还将 概括了发育中心脏发育成熟的生化和生物物理刺激的关键方面 心肌细胞。我的长期职业目标是开发生物工程方法来衍生一种 从hESCs和hiPSCs中获得足够数量的成熟心肌细胞用于心脏组织再生。这个 当前提案的目标是发展对环境影响的机械性理解 因素(例如，底物和电刺激)，意图在未来将这些信息用于茎- 基于细胞的心血管再生。这项研究具有重要意义，因为它将允许有效地推导 来自hESCs的完全成熟的心肌细胞，这可能在药物开发和心脏疾病方面产生重大影响 组织工程学。这项研究将大大受益于我的指导团队：托马斯·K·博格博士，一位 著名的发育生物学家，医学博士李久浩，一位训练有素的儿科临床医生和教职员工 MUSC儿童医院儿科心脏科成员。科布雷的核心设施将 提供从干细胞技术到研究HES细胞-材料相互作用的广泛技术支持。

Program Director/Principal Investigator (Last, First, Middle): Vyavahare Narendra R. Project summary The current inability to efficiently derive a sufficient number of mature cardiomyocytes from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hIPSCs) has severely limited the application of human stem cell technology in treating cardiovascular disease, the leading cause of death worldwide. Significant research has been conducted to engineer soluble factors, such as growth factors and small molecules, to induce cardiac differentiation of hESCs and hIPSCs. In contrast, little work has been done to optimize insoluble factors, such as the substrates on which cells grow, to facilitate cardiac differentiation. Further, the current cardiomyocytes derived from hESCs and hIPSCs are structurally and functionally similar to human embryonic/neonatal stage cardiomyocytes (i.e., immature cardiomyocytes), which have limited clinical applications. Accordingly, we will pursue two specific aims: 1) high throughput assessment of polymeric substrates for enhanced cardiac differentiation of hESCs, and 2) promote terminal differentiation of hESC- derived immature cardiomyocytes by mimicking key aspects of biochemical and biophysical stimuli in developing hearts. We hypothesize in Aim 1 that with high throughput screening of a library of polymeric substrates known to promote hESC clonal growth, substrates capable to enhance cardiac differentiation of hESCs can be identified. We hypothesize in Aim 2 that we can promote maturation of hESC-derived immature cardiomyocytes by mimicking biochemical and biophysical stimuli in developing hearts. This study is innovative: For the first time, we will utilize an emerging polymer microarray technology to develop defined substrates in a high-throughput manner to facilitate cardiac differentiation of hESCs. Further, we will recapitulate key aspects of biochemical and biophysical stimuli of developing hearts to derive mature cardiomyocytes. My long-term career goal is to develop bioengineering approaches for the derivation of a sufficient number of mature cardiomyocytes from hESCs and hIPSCs for cardiac tissue regeneration. The objective of the current proposal is to develop a mechanistic understanding of the effects of environmental factors (e.g., substrates and electrical stimulation) with the intent to use this information in the future for stem- cell based cardiovascular regeneration. The study is significant in that it would allow for efficient derivation of fully mature cardiomyocytes from hESCs, which can have major impacts in drug development and cardiac tissue engineering. The study would tremendously benefit from my mentoring team: Dr. Thomas K. Borg, a well-established developmental biologist, and Dr. Kyu-Ho Lee, MD, a trained pediatric clinician and a faculty member in the Pediatric Cardiology division at MUSC Children's Hospital. The COBRE core facilities will provide a wide range of technical support from stem cell technology to studying hES cell-materials interactions.