Inducible Notch improves progenitor cell repair of damaged heart

描述(由申请人提供)：心肌再生治疗目前受到过继转移细胞的存活和适当的谱系承诺的限制。Notch信号通路是一种在受损心肌中表达的心肌保护干细胞信号通路，在心脏干细胞的分化和向成体心肌细胞类型的分化中起着核心作用，Notch信号通路在表达水平和时间上都受到严格的调控。这项建议的长期目标是使用受调节的Notch信号来加强细胞治疗，以促进心肌修复和再生。短期目标是确定优化的Notch信号在心脏修复中的分子机制(S)。受控的Notch信号将被基因工程改造到心脏前体细胞中，以产生既可诱导又可逆转的激活的Notch信号。特定的目的将证明：1)表达干细胞特异性调节的细胞内Notch的转基因小鼠的心肌修复增加，2)表达诱导性Notch的心脏前体细胞过继转移到梗死心脏时具有增强的再生能力。目标将通过分子生物学设计来实现，即结合生化、组织学和功能分析对小鼠和人类细胞系进行基因工程。拟议的研究的意义在于获得对干细胞存活和承诺的控制，这两者共同构成了基于细胞的治疗实施的深刻限制。这种有针对性的调控Notch活性的策略不仅可以应用于心脏前体细胞，还可以应用于其他类型的干细胞，以促进再生能力并改变治疗性干细胞治疗的范式。 公共卫生相关性：心脏病仍然是美国和全世界发病率和死亡率的主要原因。最近，基于细胞的治疗心肌损伤的前景因捐献细胞的存活、植入和持久性有限而受阻。提高过继转移的心脏祖细胞的修复能力仍然是治疗实施的一个重大障碍。这项建议通过开发具有增强特性的基因工程心脏前体细胞来促进心肌再生，从而促进了心脏细胞的治疗。

DESCRIPTION (provided by applicant): Myocardial regenerative therapy is currently limited by survival and appropriate lineage commitment of adoptively transferred cells. The Notch pathway has emerged as a cardioprotective stem cell signaling pathway expressed in damaged myocardium that plays a central role in cardiac stem cell specification and differentiation to adult cardiac cell types, with Notch signaling being tightly regulated in both expression level and timing. The long-term goal of this proposal is to enhance myocardial repair and regeneration using regulated Notch signaling to enhance cellular therapy. The short-term goal is to determine molecular mechanism(s) underlying optimized Notch signaling in cardiac repair. Controlled Notch signaling will be genetically engineered into cardiac progenitor cells to produce activated Notch signaling that is both inducible and reversible. Specific aims will demonstrate: 1) Myocardial repair is increased in transgenic mice expressing stem cell-specific regulated intracellular Notch and 2) Cardiac progenitor cells engineered to express inducible Notch possess augmented regenerative capacity when adoptively transferred to infarcted hearts. Aims will be accomplished by molecular biological design to genetically engineer mouse and human cell lines in combination with biochemical, histological, and functional analyses. The significance of the proposed studies is to gain control over stem cell survival and commitment that together are profound limitations of cell-based therapeutic implementation. This targeted strategy for regulating Notch activity could be applied not only to cardiac progenitor cells but also to other stem cell types in order to promote regenerative capacity and change the paradigm for therapeutic stem cell treatment. PUBLIC HEALTH RELEVANCE: Heart disease persists as the leading cause of morbidity and mortality in the United States and throughout the world. Recent promise of cell-based therapy for treatment of cardiomyopathic injury has been stymied by limited survival, engraftment and persistence of the donated cells. Improving the reparative capacity of adoptively transferred cardiac progenitors remains a significant hurdle to therapeutic implementation. This proposal advances cardiac cell therapy through development of genetically engineered cardiac progenitors cells with enhanced properties to potentiate myocardial regeneration.