Combinatorial approaches for studying multiple cues regulating human pluripotent

描述(由申请人提供) 摘要：人多能干细胞(HPSCs)是治疗多种神经退行性疾病所致的衰弱损伤的一种很有前途的细胞治疗资源。然而，控制hPSC分化为谱系特异性神经细胞是充分发挥其神经再生医学潜力之前需要解决的最重要的问题之一。详细了解细胞外微环境和内在细胞调节因子的功能，动态地调控hPSC向神经/神经细胞的神经发生是解决上述挑战的先决条件。然而，由于我们对诱导分化的多个信号缺乏了解，以及现有的研究方法有限，hPSC微环境的功能研究要复杂得多。因此，我们的主要研究重点是开发创新的方法，以确定hPSC分化为亚型特异性神经元的最佳信号，并使用基于非病毒siRNA的转基因工具对hPSC进行遗传操作。我们的创新方法将允许建立新的基于细胞的分析工具和基于siRNA的遗传操作工具，用于选择性和高效地对hPSC进行神经分化。此外，还将努力将这些研究整合到一个多分析微流控平台中，以便协同控制微环境线索和内在细胞调节因子。该研究所在纳米生物材料、功能基因组学和干细胞生物学方面的研究经验，以及目前旨在研究微环境中细胞相互作用的跨学科研究计划，将是开发上述/创新工具的关键。 公共卫生相关性：神经退行性疾病(如阿尔茨海默病和帕金森氏病)和脊髓损伤影响着大约数百万美国人，他们因损伤而终身瘫痪甚至死亡。因此，迫切需要发展基于细胞的神经再生医学疗法，其中人类多能干细胞(HPSCs)具有独特的自我更新能力并分化为所有体细胞，是非常有前途的移植治疗资源。这项拟议研究的目标是开发创新的方法来识别和理解微环境线索和内在细胞程序对人多能干细胞(HPSC)分化为亚型特定神经元的生长、分化和分子规格的时间/空间影响。

DESCRIPTION (Provided by the applicant) Abstract: Human pluripotent stem cells (hPSCs) are promising resources as cell-based therapies for the debilitating injuries caused by many neuro-degenerative diseases. However, controlling hPSC differentiation into lineage-specific neural cells is one of the most important problems needed to be addressed before their potential for neuro-regenerative medicine can be fully realized. A detailed insight into the functions of extracellular microenvironments and intrinsic cellular regulators which dynamically regulate the hPSC neurogenesis into neural/neuronal cells is a prerequisite for addressing the aforementioned challenges. However, functions of hPSC microenvironments are much more complicated to investigate because of our lack of knowledge about the multiple signals inducing differentiation and limited methods available for investigation. Therefore, the primary focus of our study is to develop innovative methods to identify optimal cues for hPSC differentiation into subtype specific neurons and genetic manipulation of hPSCs using non-viral siRNA based transfection tools. Our innovative approaches will allow for the establishment of novel cell-based assay tools and siRNA based genetic manipulation tools for selective and efficient neuro-differentiation of hPSCs. Moreover, efforts will be made to integrate these studies into one multianalytic microfluidics platform for synchronized control of microenvironmental cues and intrinsic cellular regulators synergistically. The PI's research experiences in nanoscale biomaterials, functional genomics, and stem cell biology and current interdisciplinary research programs aiming at investigating cellular interactions within microenvironments would be critical to develop the aforementioned/innovative tools. Public Health Relevance: Neuro-degenerative diseases (e.g. Alzheimer's disease and Parkinson's disease) and spinal cord injury, affects about a few million Americans, who experience life-long debilitating paralysis or even death due to the injury. Therefore, there is an urgent need for the development of cell-based therapies for neuro-regenerative medicine, where human pluripotent stem cells (hPSCs) are extremely promising resources for transplantation therapies as they possess the unique ability to self-renew and give rise to all somatic cell lineages. Goals of this proposed study is to develop innovative methods for identifying as well as understanding the temporal/spatial effects from microenvironmental cues and intrinsic cellular programs on growth, differentiation, and molecular specification of human pluripotent stem cell (hPSC) differentiation into sub-type specific neurons.