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

该中心致力于开发鉴定和表征与细胞糖蛋白和糖脂相关的糖链结构和转录本的技术和工具。作为进一步开发糖化技术和工具的试验台，研究将再次集中在人类多能性干细胞的特征上，包括胚胎干细胞(HESCs)和诱导多能性干细胞(IPSCs)。来自干细胞的一系列分化细胞谱系将被纳入这一分析。这种生物模型是糖组学技术和工具开发的理想试验台，因为它提供了一系列不同的多糖结构，以及由于可以产生的细胞类型的多样性而带来的挑战。这些研究将为人类胚胎发育提供新的见解，并将导致识别用于干细胞表征的新生物标记物。这项研发的一个主要优势是成功地从hESC和HiPSCs中产生了纯的、特征良好的细胞类型。例如，我们例行公事地产生适合于下游分析的神经、肝脏、胰腺和心脏谱系。作为该中心上一轮资助的一部分，开发了第一种从多能细胞直接分化神经脊干细胞的方法[1]。还在开发更多的心血管谱系，以增加将由其他研发和开发人员分析的细胞的多样性，并增强将遇到的多糖复杂性。 在这一竞争性更新中，我们建议扩展我们之前的工作，并包括对糖基化相关疾病的人类IPSC模型的新强调。了解这些疾病的基础是一项重大挑战，也是一个在很大程度上未得到充分探索和了解的领域。使用IPSC重编程技术，我们建议生成完全有效的细胞系面板，这些细胞系可以作为人类先天性糖基化障碍(CDG)和先天性肌营养不良(CMD)的模型。这将为大量研究人员提供宝贵的资源，并为糖组分技术开发创造一个真正具有挑战性的试验台。这项研发的另一个主要新组成部分是纳入理查德·斯蒂特博士作为联合研究员。Steet博士将把他在先天性糖基化紊乱领域的广泛专业知识应用到这项工作中，并将主要负责对所产生的细胞系进行生化验证。 斯蒂特博士还将协调许多DBP合作者和该研发人员之间的互动。

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.