Molecular Imaging of Cardiac Pluripotent Stem Cells

项目总结 分子成像是一门新学科，它使细胞和细胞的非侵入性可视化成为可能 活体受试者的分子过程。在这里，我们将采用报告基因和报告探针成像 技术(最初是为癌症研究人员开发的)来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)。然而， 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种新的基因组编辑方法(ZFN，TALEN，CRISPR，phiC31)，它们将使 美国将安全高效地将人PET报告基因(HmTK2)导入人诱导多能性 干细胞(IPSCs)。之所以选择这些细胞类型，是因为最近发现成年体细胞 可以转化为具有无限自我更新和多潜能分化的IPSCs (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 已编辑的行。在验证后，我们将随后在体内成像IPSC-CMS在小型和大型的命运 动物心肌梗死模型以了解细胞剂量、投药时机、 和成像灵敏度。我们将通过PET报告基因成像将IPSC-CM存活率与心脏 通过核磁共振成像发挥作用。从这些研究中收集的信息应该证明 为未来将分子成像与临床干细胞治疗相结合提供了工具。

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.