TR&D1: Stem Cell and Induced Pluripotent Stem Cell Resources (Pages 116-134)

TR

• 批准号: 8708156
• 负责人: J. Michael Pierce
• 金额: $ 170.41万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8708156
• 关键词: Area; Beds; Biochemical; Biological Markers; Biological Models; Cardiac; Cardiovascular system; Cell Differentiation process; Cell Line; Cell Lineage; Cell model; Cells; Collaborations; Communities; Congenital Disorders; Development; Disease; Disease model; Embryonic Development; Enzymes; Funding; Glycobiology; Glycolipids; Glycoproteins; Human; Human Resources; Lead; Liver; Mediating; Methods; Modeling; Molecular; Neural Crest; Pancreas; Pathway interactions; Patients; Pluripotent Stem Cells; Polysaccharides; Process; Production; Quality Control; Regulation; Research Personnel; Resources; Sampling; Stem cells; Structure; Subfamily lentivirinae; Technology; Testing; Tetracyclines; Transcript; Validation; Work; base; cell type; congenital muscular dystrophy; embryonic stem cell; glycosylation; human disease; human embryonic stem cell; induced pluripotent stem cell; insight; knock-down; loss of function; overexpression; relating to nervous system; research study; small hairpin RNA; stem cell technology; success; technology development; tool; tool development

This Center is dedicated to the development of technologies and tools for the identification and characterization of glycan structures and transcripts associated with cellular glycoproteins and glycolipids. As a test-bed for the further development of glycomic technologies and tools, studies will again focus on the characterization of human pluripotent stem cells, including embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). A range of differentiated cell lineages derived from stem cells will be incorporated into this analysis. This biological model is ideal as a test-bed for glycomics technologies and tool development because it offers a diverse range of glycan structures and challenges due to the diversity in cell types that can be generated. These studies will offer new insights into human embryonic development and will lead to the identification of new biomarkers for stem cell characterization. A major strength of this TR&D has been its success in generating pure, well-characterized cell types from hESC and hiPSCs. For example, we routinely produce neural, liver, pancreatic and cardiac lineages that are suitable for downstream analysis. The first method for direct differentiation of neural crest stem cells from pluripotent cells [1] was developed as part of the Center's previous round of funding. Additional cardiovascular lineages are also being developed to increase the diversity of cells to be analyzed by other TR&Ds'and to enhance glycan complexity that will be encountered. In this competitive renewal, we propose to extend our previous work and also include a new emphasis on human iPSC models for glycosylation-related disease. Understanding the basis of these diseases represents a major challenge and an area that is largely underexplored and poorly understood. Using iPSC-reprogramming technology, we propose to generate panels of fully validated cell lines that can serve as models for human congenital disorders of glycosylation (CDG) and congenital muscular dystrophy (CMD). This will serve to provide valuable resources for a large number of investigators and to create a truly challenging test-bed for glycomics technology development. Another major new component of this TR&D is the inclusion of Dr. Richard Steet as co-investigator. Dr. Steet will bring his broad expertise in the area of congenital glycosylation disorders to this effort and will be primarily responsible for the biochemical validation of the resulting cell lines. Dr. Steet will also coordinate the interactions between many DBP collaborators and personnel within this TR&D.

该中心致力于开发用于识别和表征与细胞糖蛋白和糖脂相关的聚糖结构和转录本的技术和工具。作为进一步开发糖组学技术和工具的试验台，研究将再次集中于人类多能干细胞的表征，包括胚胎干细胞（hESC）和诱导多能干细胞（iPSC）。一系列源自干细胞的分化细胞谱系将被纳入该分析中。该生物模型非常适合作为糖组学技术和工具开发的试验台，因为它提供了多种聚糖结构，并由于可以生成的细胞类型的多样性而面临挑战。这些研究将为人类胚胎发育提供新的见解，并将导致鉴定干细胞表征的新生物标志物。这项 TR&D 的主要优势在于成功地从 hESC 和 hiPSC 中产生了纯的、特征良好的细胞类型。例如，我们通常会产生适合下游分析的神经、肝脏、胰腺和心脏谱系。第一种从多能细胞直接分化神经嵴干细胞的方法[1]是作为该中心上一轮资助的一部分而开发的。其他心血管谱系也正在开发中，以增加其他 TR&D 分析的细胞多样性，并增强将遇到的聚糖复杂性。 在这次竞争性更新中，我们建议扩展我们之前的工作，并重新强调糖基化相关疾病的人类 iPSC 模型。了解这些疾病的基础是一项重大挑战，也是一个很大程度上尚未得到充分探索和了解的领域。我们建议使用 iPSC 重编程技术生成经过充分验证的细胞系组，这些细胞系可作为人类先天性糖基化障碍 (CDG) 和先天性肌营养不良症 (CMD) 的模型。这将为大量研究人员提供宝贵的资源，并为糖组学技术开发创建一个真正具有挑战性的测试平台。该 TR&D 的另一个主要新组成部分是 Richard Steet 博士作为联合研究员。 Steet 博士将把他在先天性糖基化疾病领域的广泛专业知识带入这项工作，并将主要负责所得细胞系的生化验证。 Steet 博士还将协调该 TR&D 内许多 DBP 合作者和人员之间的互动。