Molecular Imaging of Novel Cardiomyocyte Stem Cells

新型心肌细胞干细胞的分子成像

• 批准号: 6848394
• 负责人: Steven N. Ebert
• 金额: $ 21.72万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-22 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6848394
• 关键词: bioimaging /biomedical imaging; bioluminescence; cardiac myocytes; cell differentiation; cell proliferation; embryonic stem cell; flow cytometry; gene expression; heart function; heart imaging /visualization /scanning; immunofluorescence technique; laboratory mouse; luciferin monooxygenase; magnetic resonance imaging; methyltransferase; muscle satellite cell; musculoskeletal disorder therapy; myocardial infarction; regeneration; reporter genes; stem cell transplantation; therapy design /development; tissue /cell culture

DESCRIPTION (provided by applicant): Development and application of non-invasive imaging techniques to physically and functionally assess the potential of novel cardiomyocyte stem cells to regenerate damaged myocardium have clear clinical significance. Repair and replacement of damaged cardiac muscle tissue is a key limitation in the recovery from ischemic and other forms of heart disease, the leading cause of death in the United States. New strategies for regenerating damaged cardiac muscle are urgently needed and could be readily translated to the clinical setting if first shown to be effective in animal models. The proposal outlined here will demonstrate the feasibility of using molecular imaging techniques in vivo to evaluate our novel stem cell-based strategy for cardiac regenerative medicine. This research proposal directly addresses both of the stated goals of the initiative outlined in RFA-HL-04-003 (Cellular and Molecular Imaging of the Cardiovascular, Pulmonary, and Hematopoetic Systems): (1) Detect and quantify molecular and cellular pathways that regulate heart function...(we describe and will further characterize a novel cardiomyocyte differentiation pathway), and (2) Develop new methods for cell tracking in vivo for applications in cell-based therapeutics...(we will transplant novel cardiomyocyte stem cells into damaged hearts and track them in vivo using advanced magnetic resonance and bioluminescent imaging methods). Our data indicate that cardiac cells transiently expressing the enzyme, Phenylethanolamine n-methyltransferase (Pnmt), ultimately become myocytes that contribute substantially to pacemaking, conduction, and working myocardium. Thus, Pnmt serves as a novel marker of cardiomyocyte stem cells (Pnmt+ cells) in the developing heart. We will test the idea that Pnmt+ cells can be selectively isolated and transplanted into damaged cardiac regions where they will be monitored both physically (location) and physiologically (function) using non-invasive molecular and cellular imaging techniques to evaluate the potential of these cells to regenerate cardiac muscle tissue in vivo. Three specific aims are proposed to accomplish this goal: Aim 1: Isolate and characterize Pnmt-nEGFP+ cells from cardiac-differentiated embryonic stem cells. Aim 2: In vivo magnetic resonance imaging (MRI) of transplanted cardiomyocyte stem (Pnmt+) cells. Aim 3: In vivo bioluminescence imaging (BLI) of transplanted cardiomyocyte stem (Pnmt+) cells. (End of Abstract)

描述（由申请人提供）： 开发和应用非侵入性成像技术，从物理和功能上评估新型心肌干细胞再生受损心肌的潜力，具有明确的临床意义。受损心肌组织的修复和替换是缺血性和其他形式心脏病恢复的关键限制，缺血性和其他形式心脏病是美国死亡的主要原因。迫切需要用于再生受损心肌的新策略，并且如果首次在动物模型中显示有效，则可以容易地转化为临床环境。这里概述的建议将证明在体内使用分子成像技术来评估我们的新的基于干细胞的心脏再生医学策略的可行性。这项研究提案直接解决了RFA-HL-04-003（心血管、肺和造血系统的细胞和分子成像）中概述的倡议的两个既定目标：（1）检测和量化调节心脏功能的分子和细胞通路...... (we描述并将进一步表征一种新的心肌细胞分化途径），以及（2）开发体内细胞追踪的新方法，用于基于细胞的治疗...... (we将新的心肌干细胞移植到受损的心脏中，并使用先进的磁共振和生物发光成像方法在体内跟踪它们）。我们的数据表明，心肌细胞瞬时表达的酶，苯乙醇胺N-甲基转移酶（Pnmt），最终成为心肌细胞，大大有助于起搏，传导和工作心肌。因此，Pnmt作为一种新的标记心肌细胞干细胞（Pnmt+细胞）在发育中的心脏。我们将测试Pnmt+细胞可以选择性分离并移植到受损心脏区域的想法，在那里它们将使用非侵入性分子和细胞成像技术进行物理（位置）和生理（功能）监测，以评估这些细胞在体内再生心肌组织的潜力。 为实现这一目标，提出了三个具体目标： 目的1：从心肌分化的胚胎干细胞中分离并鉴定Pnmt-nEGFP+细胞。目的2：心肌干细胞（Pnmt+）移植后的体内磁共振成像（MRI）。目的3：移植心肌干细胞（Pnmt+）的体内生物发光成像（BLI）。 (End摘要）