Cardiomyocyte-specific modified mRNA of Pkm2 for induction of cardiac regeneration in ischemic heart failure

心肌细胞特异性修饰的 Pkm2 mRNA 用于诱导缺血性心力衰竭的心脏再生

• 批准号: 10424414
• 负责人: Lior Zangi
• 金额: $ 42.38万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10424414
• 关键词: Action Potentials; Acute; Acute myocardial infarction; Address; Adult; Anisotropy; Apoptosis; Area; Arrhythmia; Biological Assay; Birth; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cause of Death; Cell Proliferation; Cells; Chronic; Cicatrix; Clinical; Cytosol; Data; Development; Dose; Down-Regulation; Electrophysiology (science); Embryonic Heart; Enzymes; Exhibits; Functional disorder; Gene Delivery; Gene Transduction Agent; Genes; Genetic Transcription; HIF1A gene; Heart; Heart Hypertrophy; Heart failure; Hypertrophy; Hypoxia; Impairment; In Vitro; Incidence; Individual; Infarction; Inflammation; Injury; Isoenzymes; Knock-out; Knockout Mice; Laboratories; Malignant - descriptor; Mediating; Membrane Potentials; Messenger RNA; Methods; Mitosis; Modeling; Molecular; Morbidity - disease rate; Muscle; Myocardial; Myocardial Ischemia; Myocardium; Myofibroblast; Natural regeneration; Nature; Neonatal; Newts; Optics; Organism; Outcome; Pathologic; Pentosephosphate Pathway; Phase; Phylogenetic Analysis; Physiologic pulse; Population; Prevention; Process; Proliferating; Property; Proteins; Pyruvate Kinase; Regenerative capacity; Resolution; Rest; Risk; Role; Secondary to; Signal Pathway; Signal Transduction; Small Interfering RNA; Therapeutic; Time; Tissue Engineering; Treatment Failure; Up-Regulation; Viral; Weight; Widow; Zebrafish; angiogenesis; base; beta catenin; c-myc Genes; cardiac regeneration; cardiac repair; clinical application; conditional knockout; design; differential expression; disability; fetal; functional loss; gene delivery system; gene therapy; genetic analysis; healing; heart function; improved; in vivo; indium arsenide; mortality; mouse model; multidisciplinary; novel; prevent; regenerative; repaired; restoration; targeted delivery; transcription factor; Action Potentials; Acute; Acute myocardial infarction; Address; Adult; Anisotropy; Apoptosis; Area; Arrhythmia; Biological Assay; Birth; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cause of Death; Cell Proliferation; Cells; Chronic; Cicatrix; Clinical; Cytosol; Data; Development; Dose; Down-Regulation; Electrophysiology (science); Embryonic Heart; Enzymes; Exhibits; Functional disorder; Gene Delivery; Gene Transduction Agent; Genes; Genetic Transcription; HIF1A gene; Heart; Heart Hypertrophy; Heart failure; Hypertrophy; Hypoxia; Impairment; In Vitro; Incidence; Individual; Infarction; Inflammation; Injury; Isoenzymes; Knock-out; Knockout Mice; Laboratories; Malignant - descriptor; Mediating; Membrane Potentials; Messenger RNA; Methods; Mitosis; Modeling; Molecular; Morbidity - disease rate; Muscle; Myocardial; Myocardial Ischemia; Myocardium; Myofibroblast; Natural regeneration; Nature; Neonatal; Newts; Optics; Organism; Outcome; Pathologic; Pentosephosphate Pathway; Phase; Phylogenetic Analysis; Physiologic pulse; Population; Prevention; Process; Proliferating; Property; Proteins; Pyruvate Kinase; Regenerative capacity; Resolution; Rest; Risk; Role; Secondary to; Signal Pathway; Signal Transduction; Small Interfering RNA; Therapeutic; Time; Tissue Engineering; Treatment Failure; Up-Regulation; Viral; Weight; Widow; Zebrafish; angiogenesis; base; beta catenin; c-myc Genes; cardiac regeneration; cardiac repair; clinical application; conditional knockout; design; differential expression; disability; fetal; functional loss; gene delivery system; gene therapy; genetic analysis; healing; heart function; improved; in vivo; indium arsenide; mortality; mouse model; multidisciplinary; novel; prevent; regenerative; repaired; restoration; targeted delivery; transcription factor

PROJECT SUMMARY Ischemic heart failure (HF) secondary to chronic post myocardial infraction (MI), is a leading cause of death and disability worldwide. The high morbidity and mortality rates in ischemic HF are caused, in large part, by the low regenerative capacity (and therefore repair) of the mammalian adult heart. Several tissue-engineering, cell- and gene-based therapies have been attempted for induction of cardiac regeneration following MI. Since cardiomyocyte (CM) proliferation and cardiac regeneration are highly robust processes in Zebrafish and Newts, efforts to mimic the pro-proliferative mechanisms of these lower phylogenetic organisms in the adult mammalian heart have been attempted using standard gene therapy vectors. Those approaches however, have encountered major challenges that are related to short-term, low, or uncontrolled delivery of the target gene. At best, this has resulted in poor CM proliferation. In other cases, these strategies have led to highly undesirable outcomes, including exacerbation of pathological hypertrophy and inflammation. We have recently shown that modified mRNA (modRNA) delivery is a safe, efficient, transient, non-immunogenic, local, and dose-controlled cardiac gene delivery platform. We have found that the glycolytic enzyme Pyruvate Kinase Muscle Isozyme M2 (Pkm2), which is highly expressed in regenerative fetal and neonatal CMs but not in adult CMs is an effective vehicle for re-awakening cardiac regeneration when delivered at the time of MI. Selective Pkm2 modRNA delivery to distinct cell populations led to a significant increase in their respective rates of proliferation. Proliferation of non-CMs such as myofibroblasts may exacerbate pathological remodeling by impairing diastolic function, disrupting myocardial conduction, and promoting arrhythmias. To circumvent this, we have designed a unique CM-specific modRNA-based approach for improving post-MI cardiac repair and regeneration. This proposal will identify the optimal therapeutic widow for modRNA-mediated delivery of Pkm2 to CMs following the phase of active healing for effective reversal of electro-mechanical dysfunction, will determine the molecular mechanisms by which local and transient delivery of Pkm2 promotes CM proliferation via β-catenin and downstream signaling, and finally determine the altered conduction properties and risk of arrhythmias associated with efficient integration of newly-formed Pkm2-dependent CMs with the host myocardium post MI. The highly translatable nature of our proposed modRNA platform and our integrative, multi-disciplinary experimental plan are ultimately geared towards clinical applicability for ischemic heart failure.

项目摘要 继发性心肌违规（MI）继发于慢性的缺血性心力衰竭（HF）是死亡的主要原因 和全世界的残疾。缺血性HF的高发病率和死亡率在很大程度上是由 哺乳动物成人心脏的低再生能力（并因此修复）。几个组织工程，细胞 - MI之后，已经尝试诱导基于基因的疗法来诱导心脏再生。自从 心肌细胞（CM）增殖和心脏再生是斑马鱼和NEW的高度稳健过程， 模仿成人这些较低系统发育生物的促生产机制的努力 使用标准基因疗法载体尝试了哺乳动物心脏。但是这些方法 遇到了与目标的短期，低或不受控制有关的主要挑战 基因。充其量，这导致CM增殖差。在其他情况下，这些策略导致了高度的 不良结果，包括加剧病理肥大和炎症。我们最近有 表明修饰的mRNA（modRNA）递送是一种安全，高效，瞬态，非免疫原性，局部和 剂量控制的心脏基因输送平台。我们发现糖酵解酶丙酮酸激酶 肌肉同工酶M2（PKM2），在再生胎儿和新生儿CM中高度表达，但在成人中不表达 CMS是在MI时交付时重新唤醒心脏再生的有效工具。选择性 PKM2 modRNA递送到不同的细胞群体导致其各自的速率显着提高 增殖。非CMS（例如肌纤维细胞）的扩散可能会通过 损害舒张功能，破坏心肌传导并促进心律不齐。为了解决这个问题， 我们设计了一种独特的CM特异性ModRNA方法，用于改善MI后心脏修复和 再生。该建议将确定用于ModRNA介导的PKM2的最佳治疗寡妇 在有效逆转电力机械功能障碍的有效逆转之后，到CMS将 确定PKM2局部和瞬态递送促进CM增殖的分子机制 通过β-catenin和下游信号传导，并最终确定传导性能的改变和风险 与新形成的PKM2依赖性CM与宿主有效整合有效整合的心律不齐 心肌后MI。我们提议的ModRNA平台和我们的集成， 多学科实验计划最终旨在用于缺血性心脏的临床适用性 失败。