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（由适用提供）：对心脏的干细胞注射是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的试验显示出适度的临床益处，但预计该田间的大幅进展是由于使用源自诱导的多能干细胞的心肌细胞而产生的。尽管有很大的希望，但最终临床使用多能干细胞衍生的心肌细胞面临许多需要解决的挑战，包括细胞成熟不足的关键问题，表型异质性，心律失常，心律失常，植入后的生存能力低，植入后的生存能力低和扩大。因此，在这个项目中，我们旨在建立一种不依赖干细胞使用的心脏细胞和基因疗法的新方法。取而代之的是，我们建议将成纤维细胞的体外或原位基因工程使用具有可自定义的电表型的电活性细胞，这些表型可以与周围的心肌细胞息息相关并改善其电气和收缩功能。具体而言，在AIM 1中，我们将利用遗传操作的最低ST来快速有效地将成年人的成纤维细胞转化为一种易于扩展且均匀的令人兴奋的细胞来源，该细胞自主发射和发挥作用潜力。在AIM 2中，具有精选电生理表型的工程成纤维细胞的特征是它们与新生大鼠的心肌细胞的功能相互作用在良好控制的体外共培养系统中。在AIM 3中，我们将确定通过植入工程的刺激成纤维细胞或内源性成纤维细胞转换为电活动细胞，是否可以改善梗塞大鼠心脏的收缩功能。除了ABOV实验研究外，我们还将利用计算机模拟来促进令人兴奋的细胞的基因工程，并增强对其在体外和体内与天然心肌细胞功能相互作用的机械理解。我们认为，拟议的遗传和组织工程方法将为未来的基于细胞和基因基因的成纤维细胞的实验和临床使用为心脏梗塞和心律不齐的疗法提供良好的基础。