Combinatorial approaches for studying multiple cues regulating human pluripotent

研究调节人类多能性的多种线索的组合方法

• 批准号: 7848757
• 负责人: Kibum Lee
• 金额: $ 231.78万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7848757
• 关键词: Address; Affect; Alzheimer' s Disease; American; Biocompatible Materials; Biological Assay; Cell Lineage; Cell Therapy; Cells; Cessation of life; Cues; Degenerative Disorder; Development; Differentiation and Growth; Goals; Human; Injury; Interdisciplinary Study; Investigation; Knowledge; Life Experience; Methods; Microfluidics; Molecular; Neurons; Paralysed; Parkinson Disease; Pluripotent Stem Cells; Regenerative Medicine; Research; Resources; Signal Transduction; Small Interfering RNA; Somatic Cell; Spinal cord injury; Transfection; Transplantation; Viral; abstracting; base; combinatorial; experience; extracellular; functional genomics; genetic manipulation; innovation; insight; nanoscale; neurogenesis; novel; programs; public health relevance; relating to nervous system; self-renewal; stem cell biology; stem cell differentiation; tool

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.

描述（由申请人提供） 摘要：人类多能干细胞（hPSC）是有前途的资源，作为基于细胞的疗法用于许多神经退行性疾病引起的衰弱性损伤。然而，控制hPSC分化为谱系特异性神经细胞是在其用于神经再生医学的潜力可以完全实现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剂的功能，其动态调节hPSC神经发生为神经/神经元细胞是解决上述挑战的先决条件。然而，hPSC微环境的功能研究要复杂得多，因为我们缺乏关于诱导分化的多种信号的知识，并且可用于研究的方法有限。因此，我们研究的主要焦点是开发创新方法来鉴定用于hPSC分化为亚型特异性神经元的最佳线索，以及使用基于非病毒siRNA的转染工具对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测定工具和基于siRNA的遗传操作工具，用于hPSC的选择性和有效的神经分化。此外，将努力将这些研究整合到一个多分析微流体平台中，以协同同步控制微环境线索和内在细胞调节因子。PI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当前旨在研究微环境中细胞相互作用的跨学科研究计划对于开发上述/创新工具至关重要。 公共卫生相关性：神经退行性疾病（例如阿尔茨海默病和帕金森病）和脊髓损伤影响大约几百万美国人，他们由于损伤而经历终身衰弱性瘫痪甚至死亡。因此，迫切需要开发用于神经再生医学的基于细胞的疗法，其中人多能干细胞（hPSC）是用于移植疗法的极有前途的资源，因为它们具有自我更新和产生所有体细胞谱系的独特能力。这项研究的目的是开发创新的方法，用于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）分化为亚型特异性神经元的生长，分化和分子特化的时间/空间效应。