Engineering a power switch to study the contribution of stem cell-derived cardiomyocytes on heart regeneration

设计电源开关来研究干细胞衍生的心肌细胞对心脏再生的贡献

基本信息

批准号：
10213825
负责人：
Emmanouil Tampakakis
金额：
$ 17.17万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-15 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10213825
关键词：
Action Potentials Address Aleurites Attention Bioinformatics Birth Calcium Calcium Channel Cardiac Cardiac Myocytes Cardiology Cardiomyopathies Cell Death Cell Line Cell Transplantation Cells Cellular biology Collaborations Core Facility Coupling Doxycycline EFRAC Electrophysiology (science)Elements Embryo Engineering Equipment Experimental Animal Model Fibroblasts Fibrosis Funding Future Gene Expression Profile Genetic Genetic Transcription Giant Cells Goals Grant Guanosine Triphosphate Phosphohydrolases Heart Heart failure Homeostasis Human Implant In Situ In Vitro Infarction Ion Channel Learning Medicine Mentors Mesenchymal Methods Microfilaments Mitochondria Modeling Molecular Monomeric GTP-Binding Proteins Morbidity - disease rate Mus Muscle Muscle Cells Myocardial Infarction Myocardial Ischemia Myocardium Names Natural regeneration Necrosis Organ Pathway interactions Patients Physicians Physiological Physiology Property Proteins Rattus Recovery of Function Research Research Personnel Scientist System Testing Tissues Training Transplantation Universities Vascularization Work cardiac regeneration cardiogenesis career career development cell type clinical application efficacy testing electrical property embryonic stem cell exosome functional improvement heart damage improved in vivo in vivo evaluation inducible gene expression insight instructor mortality novel novel strategies paracrine prevent promoter regenerative therapy regenerative treatment repaired skills stem cell biology stem cell differentiation stem cell function stem cell self renewal stem cell therapy stem cells tool transcription factor transcriptome treatment strategy voltage

项目摘要

PROJECT SUMMARY I am an instructor of Medicine at Johns Hopkins University and my goal is to become an independent physician-scientist in the field of heart regeneration with a focus on deciphering the mechanisms of stem cell regeneration therapy in damaged myocardium. Cardiac regeneration therapy holds great potential to repair damaged hearts and improve their function. In the past 15 years, several trials have tested the efficacy of various types of stem cell-based therapies in different cardiomyopathy models with varying results. A critical question that remains unanswered is whether stem cells and/or progenitor cells can differentiate into cardiomyocytes capable of establishing stable electromechanical integration with the host cardiac tissue to generate meaningful force and thus significantly improve systolic function. Alternatively, it is also possible that the beneficial effects are due to paracrine factor and exosome secretions that boost reparative pathways and prevent cell death. In this study, I propose a novel approach to directly test this fundamental question by using a conditional power switch to control the excitation and contraction, of stem cell-derived cardiomyocytes. Excitation and contraction will be controlled by conditionally expressing a GTPase named Rem1, which inhibits the voltage-activated calcium channel Cav1.2 (ICa,L), a channel essential for cardiomyocyte excitation. Rem1 under control of a doxycycline-inducible promoter was introduced into stem cells that differentiate into cardiomyocytes and upon doxycycline induction can be turned on or off. This proposal will introduce these myocytes into infarcted rat hearts and repetitively turn the power switch on and off to assess their systolic contributions. The influence of Cav1.2 inhibition by Rem1 on paracrine factor and exosome secretions, as well as transcription factor expression and mitochondria function of stem cell-derived cardiomyocytes will also be studied. Overall, this proposal will provide important insights into the mechanisms of cardiac regeneration therapy that will be critical for future refinement and widespread clinical application of this treatment. To successfully carry out this proposal, I have assembled an outstanding team. This includes leaders in cardiac physiology such as my mentor Dr. David Kass and my consultant Dr. Leslie Tung, my co-mentor Dr. Chulan Kwon an expert in heart development, and my consultants Drs. Charles Murry a world leader in cardiac regeneration and Dr. Patrick Cahan an expert in bioinformatics. They will all provide outstanding training in every method I need and will oversee and support my scientific progress and career development as an independent investigator. I already have my own lab space, lab technician and funds and I will have full access to state-of-art equipment both at the Johns Hopkins University Core facilities and in the division of Cardiology.

项目概要我是约翰·霍普金斯大学的一名医学讲师，我的目标是成为一名独立的心脏再生领域的医师科学家，专注于破译干细胞的机制受损心肌的再生治疗。心脏再生疗法具有巨大的修复潜力受损的心脏并改善其功能。在过去的15年里，多项试验测试了其功效不同类型的基于干细胞的疗法在不同的心肌病模型中具有不同的结果。一个批评的仍未得到解答的问题是干细胞和/或祖细胞是否可以分化为心肌细胞能够与宿主心脏组织建立稳定的机电一体化产生有意义的力量，从而显着改善收缩功能。或者，也有可能有益的作用是由于旁分泌因子和外泌体分泌物促进了修复途径和防止细胞死亡。在这项研究中，我提出了一种新颖的方法来直接测试这个基本问题用于控制干细胞来源的心肌细胞的兴奋和收缩的条件电源开关。兴奋和收缩将通过有条件表达名为 Rem1 的 GTPase 来控制，该酶抑制电压激活的钙通道 Cav1.2 (ICa,L)，心肌细胞兴奋所必需的通道。雷姆1 在多西环素诱导型启动子的控制下，将其引入干细胞中，并分化为心肌细胞和多西环素诱导可以打开或关闭。本提案将介绍这些将肌细胞放入梗死的大鼠心脏中，并重复打开和关闭电源开关以评估其收缩压贡献。 Rem1抑制Cav1.2对旁分泌因子和外泌体分泌的影响因为干细胞来源的心肌细胞的转录因子表达和线粒体功能也将受到影响。研究过。总的来说，该提案将为心脏再生机制提供重要的见解该疗法对于该疗法的未来改进和广泛临床应用至关重要。到成功地执行这个建议，我组建了一支优秀的团队。这包括心脏领域的领导者生理学，例如我的导师 David Kass 博士和我的顾问 Leslie Tung 博士，我的共同导师 Chulan 博士权是心脏发育方面的专家，我的顾问博士。查尔斯·默里 (Charles Murry) 心脏病领域的世界领先者再生和生物信息学专家 Patrick Cahan 博士。他们都将提供出色的培训我需要并将监督和支持我的科学进步和职业发展的每一种方法独立调查员。我已经拥有自己的实验室空间、实验室技术人员和资金，并且我将拥有完全访问权限约翰·霍普金斯大学核心设施和心脏病科配备最先进的设备。