Cell Surface Proteins in Human Cardiomyocytes

人心肌细胞的细胞表面蛋白

• 批准号: 9027643
• 负责人: Rebekah L. Gundry
• 金额: $ 34.94万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9027643
• 关键词: Adult; Affinity; Antibodies; Basic Science; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cell Surface Proteins; Cell membrane; Cell surface; Cells; Data; Development; Disease model; Electrophysiology (science); Embryo; Epitopes; Extracellular Domain; Figs - dietary; Future; Generations; Genetic; Goals; Heart; Heart Diseases; Hematopoietic stem cells; Human; Human Development; Immunophenotyping; In Vitro; Investigation; Life; Mass Spectrum Analysis; Methodology; Methods; Modeling; Modification; Monoclonal Antibodies; Pluripotent Stem Cells; Population; Proliferating; Property; Proteins; Quality Control; Reagent; Regenerative Medicine; Role; Series; Sorting - Cell Movement; Source; Staging; Surface; System; Testing; Therapeutic; Ventricular; Work; base; biomarker panel; cell type; clinically relevant; congenital heart disorder; design; drug discovery; functional outcomes; improved; induced pluripotent stem cell; innovation; insight; non-genetic; novel; novel marker; novel therapeutics; public health relevance; repaired

 DESCRIPTION (provided by applicant): The ability to isolate well-characterized human induced pluripotent stem cell derived cardiomyocytes (iCM) from mixed cell populations is a fundamental and major unfulfilled goal of cardiac regenerative medicine, disease modeling and drug discovery efforts. The overall goal of this proposal is to develop a clinically- applicable method for high-throughput isolation of purified ventricular iCM, one of the most sought after cell types for translational applications. The isolation strategy is based on immunophenotyping, wherein functional potential, identity, and isolation of clinically-relevant cells are achieved via panels of antibodies that recognize exposed surface markers. Our preliminary studies have identified more than 40 informative cell surface markers for this purpose and we have developed novel monoclonal antibodies to one novel cell surface protein not previously described in the heart. We have already determined that several of these markers are restricted to specific chambers in the heart, indicating they will be highly informative for sorting subtype-specific iCM in vitro. The work will be carried out in two Aims designed to test the hypothesis that antibodies to cell surface proteins can be used to isolate ventricular iCM (Aim 1) and to determine their functional role in ventricular cardiomyogenesis (Aim 2). This work will contribute a new, non-transgene based high-throughput method for isolating live ventricular cardiomyocytes, which is not possible by current methodologies. The significance of this proposal lies with the functional outcomes enabled by the technological developments, as when an informative marker panel for sorting stage and subtype specific cells is developed, this will enable the reproducible isolation of ventricular iCM suitable for drug-discovery, therapeutic, and disease modeling studies.

 描述（由适用提供）：从混合细胞种群中分离出良好的人类诱导的多能干细胞衍生的心肌细胞（ICM）的能力是心脏再生医学，疾病建模和药物发现工作的基本和主要未实现的目标。该建议的总体目标是开发一种可临床适用的方法，用于高通量隔离纯化的心室ICM，这是最受欢迎的细胞之一 翻译应用的类型。隔离策略是基于免疫型的，其中通过临床上相关细胞的功能潜力，身份和隔离是通过 识别裸露表面标记的抗体面板。我们的初步研究为此目的确定了40多个信息的细胞表面标记，我们已经开发了一种新型的单克隆抗体，该抗体针对一种先前在心脏中未描述的新型细胞表面蛋白。我们已经确定其中一些标记仅限于心脏中的特定腔室，这表明它们在体外对亚型特异性ICM进行分类非常有用。这项工作将在两个目的中进行，旨在测试以下假设：对细胞表面蛋白的抗体可用于分离心室ICM（AIM 1）并确定其在心室心肌生成中的功能作用（AIM 2）。这项工作将贡献一种基于非转移的高通量方法，用于隔离活心房心肌细胞，这是通过当前方法不可能的。该提案的重要性在于技术发展的功能结果，因为当开发了用于分类阶段和亚型特异性细胞的信息标记面板时，这将使可重复的心室ICM可复制分离，适合用于药物分离，治疗和疾病建模研究。