Harnessing Glycoproteomics and Glycomics to Understand Cardiac Biology and Disease

摘要 我们的研究计划开发和应用创新的质谱学技术、生物信息学工具、 和方法来改变我们对细胞表面蛋白和多糖的理解，并回答杰出的问题 干细胞生物学和心脏病理学中的问题。我们的分析平台促进了新的 提高干细胞来源心肌细胞质量和均质性的试剂和策略 以及临床应用和发现监测和治疗晚期心力衰竭患者的策略。 具体地说，我们的方法能够识别、表征和量化细胞表面 从少量的人类细胞中提取的糖蛋白和多糖。到目前为止，我们已经开发了新的标记 高特异性心肌细胞的鉴定及成熟期特异性干细胞的筛选 心肌细胞，以实现对功能定义细胞的可重复评估和分离。应用我们的 技术来原代人类心脏组织，我们已经开始开发细胞类型的细胞表面的特定视图 正常心脏和衰竭心脏中的蛋白质组和糖蛋白。我们还开发了创新的生物信息学工具 告知我们下一阶段的技术发展，以增强我们的能力，提高速度和 我们分析质谱学数据的精确度。拟议的研究建立在这些经验的基础上 为了：1)开发下一代技术，它将提供关于分子的无与伦比的特异性 出现在细胞表面的表型，这是目前任何方法都不可能获得的信息，2) 开发标记面板，以评估和选择特定的小室和成熟阶段 干细胞来源的心肌细胞不需要基因编辑，3)定义细胞类型的特异性受体，膜- 正常人心脏中存在的结合配体和分泌因子及其在疾病中如何转变为 提供对细胞间信号的新理解，并为遥感标记的发展提供信息 有利于晚期心力衰竭患者的护理。拟议研究的影响将在未来发生。 基于我们产生的方法和数据，将可能进行的应用和机制研究。 随着我们的计划的发展，我们对我们揭示的分子进行机械和翻译研究 我们预计，这些研究的结果将广泛影响以下战略的制定 提高干细胞衍生品的质量，以促进其在药物测试、疾病建模和 治疗应用，为细胞型定向有效载荷输送系统和药物的开发提供信息 避免心脏毒性效应，并产生新的策略来评估和治疗晚期心力衰竭。

ABSTRACT Our research program develops and applies innovative mass spectrometry technologies, bioinformatics tools, and methodologies to transform our understanding of cell surface proteins and glycans and answer outstanding questions in stem cell biology and cardiac pathology. Our analytical platforms promote the development of new reagents and strategies to improve the quality and homogeneity of stem cell-derived cardiomyocytes for research and clinical applications and the discovery of strategies to monitor and treat patients with advanced heart failure. Specifically, our approaches enable the identification, characterization, and quantification of cell surface glycoproteins and glycans from small numbers of human cells. To date, we have developed new markers for identifying cardiomyocytes with high specificity and selecting maturation stage-specific stem cell derived cardiomyocytes to enable reproducible assessment and isolation of functionally-defined cells. Applying our technologies to primary human heart tissue, we have begun to develop cell-type specific views of the cell surface proteome and glycome within normal and failing hearts. We have also developed innovative bioinformatics tools to inform our next level of technology development to enhance our capabilities and improve the speed and accuracy with which we analyze our mass spectrometry data. The proposed studies build on these experiences to: 1) develop the next generation technology that will provide unparalleled specificity regarding the molecular phenotypes presented at the cell surface, information that is not possible to obtain by any current method, 2) develop marker panels that enable the assessment and selection of chamber- and maturation-stage specific stem cell derived cardiomyocytes without genetic editing, 3) define the cell-type specific receptors, membrane- bound ligands and secreted factors present in the normal human heart and how they change in disease to provide new understanding of intercellular signaling and inform the development of remote sensing markers to benefit the care of patients with advanced heart failure. The impact from the proposed studies lies within future applications and mechanistic studies that will be possible based on the approaches and data that we generate. As our program evolves to pursue mechanistic and translational studies of the molecules revealed by our discovery efforts, we expect the outcomes of these studies will broadly impact the development of strategies to improve the quality of stem cell derivatives to promote their utility for drug testing, disease modeling, and therapeutic applications, inform the development of cell-type directed payload delivery systems and drugs that avoid cardiotoxic effects, and yield new strategies to assess and treat advanced heart failure.