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技术，生物信息学工具， 和方法来改变我们对细胞表面蛋白质和聚糖的理解， 干细胞生物学和心脏病理学的问题。我们的分析平台促进新的 提高研究用干细胞衍生心肌细胞的质量和均一性的试剂和策略 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。 具体来说，我们的方法能够识别，表征和定量细胞表面 糖蛋白和聚糖从少量的人类细胞。到目前为止，我们已经开发了新的标记物， 以高特异性鉴定心肌细胞并选择成熟阶段特异性干细胞衍生的 心肌细胞，以实现功能限定的细胞的可再现的评估和分离。应用我们 通过将这些技术应用于初级人类心脏组织，我们已经开始开发细胞表面的细胞类型特异性视图 正常和衰竭心脏中的蛋白质组和糖组。我们还开发了创新的生物信息学工具 通知我们的下一个技术发展水平，以提高我们的能力和提高速度， 我们分析质谱数据的准确性。拟议的研究是在这些经验的基础上进行的 目的：1）开发下一代技术，提供无与伦比的分子特异性， 在细胞表面呈现的表型，通过任何现有方法都不可能获得的信息，2） 开发标记面板，以便评估和选择特定的腔室和成熟阶段 没有基因编辑的干细胞衍生的心肌细胞，3）定义细胞类型特异性受体，膜- 结合配体和分泌因子存在于正常人的心脏，以及他们如何改变疾病， 提供对细胞间信号传导的新理解，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。拟议研究的影响在于未来 基于我们生成的方法和数据，应用和机理研究将成为可能。 随着我们的计划的发展，我们将继续对我们发现的分子进行机械和翻译研究。 发现的努力，我们预计这些研究的结果将广泛影响战略的发展， 提高干细胞衍生物的质量，以促进其在药物测试、疾病建模和 治疗应用，告知细胞类型定向有效载荷递送系统和药物的开发， 避免心脏毒性作用，并产生评估和治疗晚期心力衰竭的新策略。

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.