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

• 批准号: 9978102
• 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
• 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/9978102
• 关键词: Action Potentials; Address; Aleurites; Attention; Bioinformatics; Birth; Calcium; Calcium Channel; Cardiac; Cardiac Myocytes; Cardiology; Cardiomyopathies; Cell Death; Cell Line; Cell Therapy; 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; regenerative therapy; repaired; skills; stem cell biology; stem cell differentiation; stem cells; tool; transcription factor; transcriptome; treatment strategy; voltage

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.

项目摘要 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 心脏再生领域的医生-科学家，专注于破译干细胞的机制 损伤心肌的再生治疗。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，有几项试验测试了 不同类型的基于干细胞的治疗在不同的心肌病模型中具有不同的结果。一个关键 仍然没有答案的问题是干细胞和/或祖细胞是否可以分化为 能够与宿主心脏组织建立稳定的机电整合的心肌细胞， 产生有意义的力，从而显著改善收缩功能。或者，也可能的是， 有益的作用是由于旁分泌因子和外泌体分泌物促进修复途径， 防止细胞死亡。在这项研究中，我提出了一种新的方法来直接测试这个基本问题， 一种条件电源开关，用于控制干细胞衍生的心肌细胞的兴奋和收缩。 兴奋和收缩将通过有条件地表达一种名为Rem 1的GT3来控制， 电压激活的钙通道Cav1.2（伊卡，L），心肌细胞兴奋所必需的通道。Rem1 在多西环素诱导型启动子的控制下， 心肌细胞和多西环素诱导后可以打开或关闭。本提案将介绍这些 心肌细胞进入梗死大鼠心脏，并反复打开和关闭电源开关，以评估其收缩压。 捐款. Rem 1抑制Cav1.2对旁分泌因子和外泌体分泌的影响 因为干细胞衍生的心肌细胞的转录因子表达和线粒体功能也将被 研究了总的来说，这一提议将为心脏再生机制提供重要的见解 这对于该治疗的未来改进和广泛临床应用将是至关重要的。到 我成功地执行了这项建议，组建了一个出色团队。这包括心脏病领域的领导者 我的导师大卫卡斯博士和我的顾问莱斯利董博士，我的共同导师朱兰博士 Kwon是心脏发育方面的专家，我的顾问Charles Murry博士是心脏发育方面的世界领先者。 再生和帕特里克卡汉博士在生物信息学专家。他们都将提供出色的培训， 我需要的每一种方法，并将监督和支持我的科学进步和职业发展， 独立调查员我已经有了自己的实验室空间、实验室技术人员和资金， 约翰霍普金斯大学核心设施和心脏病学部门的最先进设备。