Role of the NO-CO Module in Regulating CPC Function

心脏祖细胞（cardiac progenitor cells，CPCs）的发现为心脏病的治疗提供了新的途径 我们首次在HF患者中进行的CPC人体临床研究（SCIPIO）的初步结果如下： 取得了令人鼓舞然而，基于CPC的治疗受到几乎所有（至少97%） 移植的CPC在移植后不久死亡。这意味着，提高生存率， 预防移植细胞凋亡将提高CPC的治疗效果。在目前的资金 在本计划项目期间，我们发现一氧化碳（CO）和一氧化氮（NO） 强大的抗凋亡作用，并形成一个密切相关的功能模块（CO-NO模块）， 受血红素氧合酶-1（HO-1）、细胞外超氧化物歧化酶（ecSOD）和诱导型NO的调节 合成酶（iNOS）。我们将利用这一发现来增强基于细胞的疗法。项目的总体目标 目的：1.阐明CO-NO模块在CPC功能调节中的作用，评价其治疗CPC的作用 心肌梗死后的效用（MI）。我们建议，增加这个模块将大大增强 移植CPC的有效性和显著增强CPC介导的心脏修复。这个项目 代表了我们以前在本计划项目中的工作的自然演变;发现CO- NO模块对心肌缺血提供了强有力的保护，我们现在将在这项工作的基础上， 增强CPC的修复能力。在目的1中，我们将确定HO-1和CO在调节 CPC功能。目的2：探讨ecSOD在调节CPC功能和介导HO-1- 2表达中的作用。 CPCs的诱导保护。在目标3中，我们将确定NO在调节CPC功能中的作用，以及NO在CPC中的作用。 HO-1和ecSOD在介导NO对CPC影响中的作用。同时使用功能增益和功能损失 在所有三个目标中，我们将系统地评估CPC能力的基本参数， 体外和CPC在MI后LV重构的小鼠模型中修复心脏损伤的能力 和功能障碍。目标4.我们将阐明CO和NO上调的分子机制， NO上调CPC中HO-1和ecSOD，主要作用于转录因子Nrf 2。这些将 是第一个研究CO和NO的作用，以及它们的支持蛋白HO-1，ecSOD， INOS在调节CPC功能中的作用。结果将是全新的，并将增加一个新的层面， 了解CPC生物学。此外，这些研究将为治疗提供原理证明。 在CPC中加强CO-NO模块的操纵的实用性，这可能为将来的研究奠定基础。 在HF患者中进行的基因增强型或非基因增强型CPC试验。 相关性（参见说明）： 2009年，我们开始了SCIPIO，一项关于心力衰竭患者c-kit+心脏祖细胞（CPC）的研究， 初步结果令人鼓舞。项目1基于SCIPIO。我们将测试一种新疗法-基因疗法- 修饰或预处理的CPC，其承诺上级目前在SCIPIO中使用的细胞。 如果结果证实了我们的工作假设，这些研究将为更有效的测试奠定基础。 CPC用于治疗心力衰竭，因此可能有助于将细胞疗法转化为人类。

The discovery of cardiac progenitor cells (CPCs) provides a potential new approach to the treatment of heart failure (HF), The initial results of our first-in-humans clinical study of CPCs in patients with HF (SCIPIO) are encouraging. However, CPC-based therapies are severely limited by the fact that almost all (at least 97%) of the transplanted CPCs die shortly after transplantafion. This implies that increasing the survival of transplanted cells by prevenfing apoptosis will enhance the efficacy of CPC therapy. In the current funding period of this Program Project, we have found that carbon monoxide (CO) and nitric oxide (NO) exert powerful anti-apoptotic actions and form a closely inter-related functional module (CO-NO module), which is regulated by heme oxygenase-1 (HO-1), extracellular superoxide dismutase (ecSOD), and inducible NO synthase (iNOS). We will exploit this discovery to enhance cell-based therapies. The overall goal of Proiect 1 is to elucidate the role of the CO-NO module in regulating CPC function and to evaluate its therapeutic utility after myocardial infarction (Ml). We propose that augmenting this module will greatly potentiate the effectiveness of transplanted CPCs and dramaticallv enhance CPC-mediated cardiac repair. This Project represents the natural evolution of our previous work in this Program Project; having discovered that the CO- NO module affors powerful protection against mmyocardial ischemia, we will now build on this work to enhance the reparative ability of CPCs. In Aim 1, we will determine the roles of HO-1 and CO in modulating CPC function. In Aim 2, we will establish the role of ecSOD in regulating CPC function and mediating HO-1- induced protection of CPCs. In Aim 3, we will determine the role of NO in modulating CPC function and the role of HO-1 and ecSOD in mediating the effects of NO on CPCs. Using both gain- and loss-of-function approaches, in all three Aims we will systematically evaluate fundamental parameters of CPC competence in vitro and the ability of CPCs to repair cardiac damage in vivo in a murine model of post-MI LV remodeling and dysfunction. In Aim 4. we will elucidate the molecular mechanisms whereby CO and NO upregulate ecSOD, and NO upregulates HO-1 and ecSOD in CPCs, focusing on the transcription factor Nrf2. These will be the first studies to examine the role of CO and NO, and their supporting proteins HO-1, ecSOD, and INOS, in modulating CPC function. The results will be entirely new and will add a new dimension to our understanding of CPC biology. In addition, these studies will provide proof-of-principle for the therapeutic utility of manipulations that potentiate the CO-NO module in CPCs, which may lay the groundwork for future trials of genetically or pharmacologically enhanced CPCs in patients with HF. RELEVANCE (See instructions): In 2009 we started SCIPIO, a study of c-kit+ cardiac progenitor cells (CPCs) in patients with heart failure, and the initial results are encouraging. Project 1 builds on SCIPIO. We will test a new therapy - genetically- modified or preconditioned CPCs, which promise to be superior to the cells currently being used in SCIPIO. If the results confirm our working hypothesis, these studies will lay the groundwork for testing more effective CPCs for the treatment of heart failure and thus may facilitate translation of cell therapies to humans.