Polymer Microarrays for Stem Cell Cardiac Differentiation

项目主任/主要研究者（最后，第一，中间）：Vyavahare Narendra R. 项目摘要 目前无法有效地从人胚胎中获得足够数量的成熟心肌细胞， 干细胞（hESC）和人诱导多能干细胞（hIPSC）的应用已经严重限制了 人类干细胞技术用于治疗心血管疾病，这是全球死亡的主要原因。 已经进行了重要的研究来工程化可溶性因子，例如生长因子和小分子生物学因子。 分子，以诱导hESC和hIPSC的心脏分化。相比之下， 优化不溶性因子，如细胞生长的基质，以促进心脏分化。 此外，目前来源于hESC和hIPSC的心肌细胞在结构和功能上与 人胚胎/新生期心肌细胞（即，不成熟的心肌细胞），其具有有限的临床 应用.因此，我们将追求两个具体的目标：1）高通量评估聚合物 用于增强hESC的心脏分化的底物，和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， 发展心脏。我们在目标1中假设，通过高通量筛选聚合物文库， 已知促进hESC克隆生长的底物，能够增强hESC的心脏分化的底物， 可以鉴定hESC。我们在目标2中假设，我们可以促进hESC衍生的未成熟的 通过模拟发育中心脏的生物化学和生物物理刺激来刺激心肌细胞。本研究 创新：我们将首次利用新兴的聚合物微阵列技术来开发定义明确的 本发明涉及以高通量的方式在底物中诱导hESC分化，以促进hESC的心脏分化。此外，我们将 概括发育心脏的生化和生物物理刺激的关键方面，以获得成熟的 心肌细胞我的长期职业目标是开发生物工程方法， 足够数量的来自hESC和hIPSC的成熟心肌细胞用于心脏组织再生。的 本提案的目标是对环境影响的机械理解， 因素（例如，基质和电刺激），目的是将来将此信息用于股骨柄， 基于细胞的心血管再生。这项研究的意义在于它将允许有效地推导出 来自hESC的完全成熟的心肌细胞，这可能对药物开发和心脏功能产生重大影响。 组织工程学这项研究将极大地受益于我的指导团队：托马斯K。博格a 一位成熟的发育生物学家，和Kyu-Ho Lee博士，医学博士，一位训练有素的儿科临床医生和一位教师， MUSC儿童医院儿科心脏科的成员。COBRE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.