Molecular Imaging of Cardiac Pluripotent Stem Cells

心脏多能干细胞的分子成像

• 批准号: 9478345
• 负责人: Joseph C. Wu
• 金额: $ 47.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-21 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9478345
• 关键词: Acute; Address; Adopted; Adult; Adverse effects; Animal Model; Animals; Autopsy; Basic Science; Biodistribution; Biological; Blood Circulation; CD34 gene; Cardiac; Cardiac Myocytes; Cardiology; Cell Count; Cell Cycle Kinetics; Cell Death; Cell Line; Cell Survival; Cell Therapy; Cell Transplants; Cells; Chromosomes; Chronic; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Coronary Arteriosclerosis; Development; Dilated Cardiomyopathy; Discipline; Drug Kinetics; Engraftment; Family suidae; Future; Generations; Genetic Transcription; Genomics; Goals; Grant; Heart failure; Histologic; Histology; Human; Image; Imagery; Imaging Techniques; Inflammation; Injections; Integrase; Ischemia; Journals; Location; Magnetic Resonance Imaging; Malignant Neoplasms; Medicine; Methods; Mitochondria; Modeling; Molecular; Monitor; Morbidity - disease rate; Multimodal Imaging; Myocardial; Myocardial Infarction; Myocardial Ischemia; Paper; Pathologic Processes; Pathology; Pathway interactions; Patients; Phenotype; Physics; Pluripotent Stem Cells; Positron-Emission Tomography; Principal Investigator; Process; Program Description; Progressive Disease; Proliferating; Publishing; Regenerative Medicine; Reporter; Reporter Genes; Research Personnel; Safety; Simplexvirus; Somatic Cell; Stem cell transplant; Stem cells; Techniques; Technology; Therapeutic; Thymidine Kinase; Translational Research; Transplantation; Treatment Efficacy; Validation; base; cell motility; cell type; cellular imaging; clinical imaging; clinical translation; dosage; experience; functional outcomes; genome editing; heart damage; heart function; heart imaging; human embryonic stem cell; image guided; imaging probe; immunogenicity; immunosuppressed; improved; in vivo; in vivo imaging; induced pluripotent stem cell; insight; inter-individual variation; interdisciplinary approach; molecular imaging; mortality; new technology; novel; novel strategies; prevent; programs; response; screening; self-renewal; spatiotemporal; stem cell biology; stem cell therapy; transcription activator-like effector nucleases; zinc finger nuclease

PROJECT SUMMARY Molecular imaging is a new discipline that makes possible the noninvasive visualization of cellular and molecular processes in living subjects. Here we will adopt the reporter gene and reporter probe imaging technique (developed initially for cancer researchers) to solve a different important problem in cardiology (i.e., understand pharmacokinetics and biodistribution of cardiac stem cell transplantation). However, the Achilles’ heel of reporter gene imaging has always been random integration into cellular chromosomes. Here we will develop 4 novel genome editing approaches (ZFN, TALEN, CRISPR, phiC31) that will enable us to safely and efficiently introduce human PET reporter gene (hmTK2) into human induced pluripotent stem cells (iPSCs). These cell types are chosen because of the recent discovery that adult somatic cells can be transformed into iPSCs that acquire both unlimited self-renewal and pluripotent differentiation (similar to human embryonic stem cells). We will perform comprehensive characterization of these genome edited lines. Upon validation, we will subsequently image in vivo fate of iPSC-CMs in both small and large animal myocardial infarction models to understand the biological effects of cell dosage, timing of delivery, and imaging sensitivity. We will correlate iPSC-CM survival by PET reporter gene imaging and cardiac function by magnetic resonance imaging. The information gathered from these studies should prove instrumental for marrying molecular imaging with clinical stem cell therapy in the future.

项目概要 分子成像是一门新学科，它使细胞和细胞的无创可视化成为可能。 在这里，我们将采用报告基因和报告探针成像。 技术（最初为癌症研究人员开发）解决心脏病学中的另一个重要问题 （即了解心脏干细胞移植的药代动力学和生物分布）。 报告基因成像的致命弱点一直是随机整合到细胞染色体中。 在这里，我们将开发 4 种新颖的基因组编辑方法（ZFN、TALEN、CRISPR、phiC31），这将使 我们安全高效地将人类PET报告基因（hmTK2）引入人类诱导多能性 选择这些细胞类型是因为最近发现成体体细胞。 可转化为具有无限自我更新和多能分化能力的 iPSC （类似于人类胚胎干细胞）我们将对这些基因组进行全面的表征。 经过验证后，我们将随后对小型和大型 iPSC-CM 的体内命运进行成像。 动物心肌梗死模型，以了解细胞剂量、递送时间、 我们将通过 PET 报告基因成像和心脏将 iPSC-CM 存活率关联起来。 从这些研究中收集的信息应该可以证明磁共振成像的功能。 有助于未来将分子成像与临床干细胞治疗结合起来。