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Engineering of Human Excitable Tissues from Unexcitable Cells

从不可兴奋细胞改造人类可兴奋组织

基本信息

批准号：
9046968
负责人：
Nenad Bursac
金额：
$ 44.56万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-15 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9046968
关键词：
Action Potentials Adult Aftercare Algorithms Animals Arrhythmia Biomedical Engineering Cardiac Cardiac Myocytes Cell Maturation Cell Therapy Cells Clinical Coculture Techniques Computer Simulation Connexin 43 Coupling Derivation procedure Dermal Disease Echocardiography Electrophysiology (science)Engineering Face Fibroblasts Fire - disasters Foundations Future Genes Genetic Genetic Engineering Healed Heart Heart Atrium Heart Diseases Heart failure Heterogeneity Human Human Engineering In Situ In Vitro Incidence Infarction Injection of therapeutic agent Label Maps Measurement Membrane Potentials Methods Modeling Myocardial Infarction Neonatal Nodal Optics Outcome Phenotype Pluripotent Stem Cells Potassium Channel Process Production Protocols documentation Publishing Rattus Reporter Retroviridae Sodium Channel Source Stem cells Surgical sutures System Techniques Testing Therapeutic Time Tissue Engineering Tissues Translations Transplantation Ventricular base cardiac repair cell type cellular engineering design electrical property engineering design functional outcomes gene therapy genetic manipulation healing implantation improved in vitro Assay in vivo induced pluripotent stem cell novel novel strategies overexpression preclinical study pressure public health relevance research study scale up screening sensor tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Stem cell injections into the heart are actively being pursued as a potential therapy for myocardial infarction and heart failure. While the ongoing trials with adult-derived stem cells show moderate clinical benefits, significant progress in the field is expected to arise from the use of cardiomyocytes derived from induced pluripotent stem cells. Despite great promise, eventual clinical use of pluripotent stem cell-derived cardiomyocytes faces a number of challenges that need to be resolved including key issues with inadequate cell maturation, phenotypic heterogeneity, arrhythmogenesis, low viability after implantation, and scale-up. Therefore, in this project we aim to establish a novel approach for cardiac cell and gene therapy that does not rely on the use of stem cells. Instead we propose to employ in vitro or in situ genetic engineering of fibroblasts into electrically active cells with customizable electrical phenotype that can couple with surrounding cardiomyocytes and improve their electrical and contractile function. Specifically, in Aim 1 we will utilize minimum st of genetic manipulations to rapidly and efficiently convert adult human fibroblasts into a readily expandable and homogeneous source of excitable cells that autonomously fire and conduct action potentials. In Aim 2, engineered fibroblasts with select electrophysiological phenotypes will be characterized for their functional interactions with neonatal rat cardiomyocytes in well-controlled in vitro co- culture systems. In Aim 3, we will establish if contractile function of infarcted rat hearts can be improved by implantation of engineered excitable fibroblasts or retroviral conversion of endogenous fibroblasts into electrically active cells. In addition to abov experimental studies, we will utilize computer simulations to facilitate genetic engineering of excitable cells and enhance mechanistic understanding of their functional interactions with native cardiomyocytes in vitro and in vivo. We believe that the proposed genetic and tissue engineering approach will provide strong foundation for the future experimental and clinical use of engineered fibroblasts in cell- and gene-based therapies for cardiac infarction and arrhythmias.

描述（由申请人提供）：正在积极寻求将干细胞注射到心脏中作为心肌梗塞和心力衰竭的潜在疗法。虽然正在进行的成人干细胞试验显示出一定的临床益处，但使用诱导多能干细胞衍生的心肌细胞预计会在该领域取得重大进展。尽管前景广阔，但多能干细胞来源的心肌细胞的最终临床应用面临着许多需要解决的挑战，包括细胞成熟不足、表型异质性、心律失常、植入后活力低和扩大规模等关键问题。因此，在这个项目中，我们的目标是建立一种不依赖于干细胞使用的心脏细胞和基因治疗的新方法。相反，我们建议采用体外或原位基因工程将成纤维细胞转化为具有可定制电表型的电活性细胞，这些细胞可以与周围的心肌细胞耦合并改善其电和收缩功能。具体来说，在目标 1 中，我们将利用最少的遗传操作来快速有效地将成人成纤维细胞转化为易于扩展且均质的可兴奋细胞源，这些细胞可自主激发和传导动作电位。在目标 2 中，将表征具有选定电生理表型的工程成纤维细胞在良好控制的体外共培养系统中与新生大鼠心肌细胞的功能相互作用。在目标 3 中，我们将确定是否可以通过植入工程化的可兴奋成纤维细胞或将内源成纤维细胞逆转录病毒转化为电活性细胞来改善梗塞大鼠心脏的收缩功能。除了上述实验研究之外，我们将利用计算机模拟来促进可兴奋细胞的基因工程，并增强对其与体外和体内天然心肌细胞功能相互作用的机制理解。我们相信，所提出的基因和组织工程方法将为工程成纤维细胞在基于细胞和基因的心肌梗塞和心律失常治疗中的未来实验和临床应用提供坚实的基础。