Chemical editing of nanoscale proteoglycan architecture and organization to control cellular

化学编辑纳米级蛋白聚糖结构和组织以控制细胞

• 批准号: 9815714
• 负责人: Mia L Huang
• 金额: $ 4.52万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9815714
• 关键词: Architecture; Binding; Carbohydrates; Cell Compartmentation; Cell membrane; Cell surface; Chemicals; Complex; Core Protein; Development; Engineering; Glycoconjugates; Glycosaminoglycans; Goals; Growth Factor; Human Development; Knowledge; Lead; Length; Link; Maps; Membrane; Methods; Monitor; Mus; Musculoskeletal; Natural regeneration; Outcome; Protein Engineering; Proteoglycan; Regenerative Medicine; Research; Signal Transduction; Site; Skeletal Muscle; Skeletal Muscle Satellite Cells; Stem cells; Structure; Techniques; To specify; cellular development; embryonic stem cell; glycosylation; nanoscale; polysulfated glycosaminoglycan; programs; proteoglycan core protein; stem cell differentiation

PROJECT SUMMARY Cellular development is controlled by cell surface proteoglycans, a class of glycoconjugates that can orchestrate the binding and release of growth factors to initiate differentiation or prime stem cells towards specific lineages. Proteoglycans are attractive targets for exerting exogenous control to re-program cellular differentiation, and yet despite their potential for biomedicine, strategies that harness their regulatory functions are limited. This gap is caused by the lack of techniques that recognize their chemical and nanoscale complexity. Proteoglycans are composed of a core protein covalently linked to a highly complex, heterogenous mixture of sulfated glycosaminoglycan carbohydrates. Furthermore, proteoglycans can be located in distinct compartments of the cell membrane, and this distribution can lead to different outcomes in differentiation. This application outlines the development of chemical strategies to mimic and probe the nanoscale architecture and organization of proteoglycans in cellular development. In two specific aims, we will demonstrate that the techniques developed through this proposal will enable new chemical strategies to control stem cell differentiation and the regeneration of skeletal muscles. First, we will develop protein engineering strategies that facilitate the bioorthogonal conjugation of semi-synthetic glycosaminoglycans to specified sites along the proteoglycan core protein chain, in order to generate "engineered" cell surface proteoglycans with "pre-mapped" glycosylation sites, as well as defined glycosaminoglycan compositions and lengths. Second, we will develop chemical methods to direct and monitor the membrane localization of such "engineered" proteoglycans. Both strategies will be applied in order to study and to exert control over the differentiation of mouse embryonic stem cells and the activation of skeletal satellite muscle cells. The long-term goal of this proposal is to offer strategies to alter proteoglycan structure in order to manipulate cellular differentiation, and to harness this knowledge towards understanding the factors that regulate canonical human development.

项目概要 细胞发育由细胞表面蛋白聚糖控制，这是一类可以协调的糖复合物 生长因子的结合和释放以启动分化或将干细胞引导至特定谱系。 蛋白多糖是施加外源控制以重新编程细胞分化的有吸引力的靶标，但 尽管它们在生物医学方面具有潜力，但利用其调节功能的策略仍然有限。这个差距是 这是由于缺乏识别其化学和纳米级复杂性的技术造成的。蛋白多糖是 由与高度复杂、异质的硫酸化混合物共价连接的核心蛋白组成 糖胺聚糖碳水化合物。此外，蛋白多糖可以位于不同的区室中。 细胞膜，这种分布会导致分化的不同结果。该应用程序概述了 开发化学策略来模拟和探测纳米级结构和组织 细胞发育中的蛋白多糖。在两个具体目标中，我们将证明所开发的技术 通过该提案将使新的化学策略能够控制干细胞的分化和再生 骨骼肌。首先，我们将开发促进生物正交的蛋白质工程策略 将半合成糖胺聚糖缀合至蛋白聚糖核心蛋白链上的特定位点， 为了生成具有“预先映射”糖基化位点的“工程化”细胞表面蛋白聚糖，以及 定义的糖胺聚糖组成和长度。其次，我们将开发化学方法来指导和 监测这种“工程化”蛋白聚糖的膜定位。两种策略将按顺序应用 研究并控制小鼠胚胎干细胞的分化和骨骼肌的激活 卫星肌细胞。该提案的长期目标是提供改变蛋白聚糖结构的策略 为了操纵细胞分化，并利用这些知识来理解影响细胞分化的因素 规范人类发展。