Role of the NO-CO Module in Regulating CPC Function

心脏祖细胞（CPCs）的发现为心力衰竭（HF）的治疗提供了一种潜在的新方法，我们首次在心力衰竭（SCIPIO）患者中进行的CPCs临床研究的初步结果令人鼓舞。然而，几乎所有（至少97%）移植的心肌细胞在移植后不久死亡，这严重限制了基于心肌细胞的治疗。这表明通过防止细胞凋亡来提高移植细胞的存活率可以提高CPC治疗的疗效。在本项目当前资助期内，我们发现一氧化碳（CO）和一氧化氮（NO）具有强大的抗凋亡作用，并形成一个密切相关的功能模块（CO-NO模块），该功能模块由血红素加氧酶-1 （HO-1）、细胞外超氧化物歧化酶（ecSOD）和诱导型NO合成酶（iNOS）调节。我们将利用这一发现来加强基于细胞的治疗。项目1的总体目标是阐明CO-NO模块在调节CPC功能中的作用，并评估其在心肌梗死（Ml）后的治疗作用。我们建议增加这个模块将大大增强移植的心肌细胞的有效性，并显著增强心肌细胞介导的心脏修复。本项目代表了我们在本项目中以前工作的自然演变；在发现CO- NO模块对心肌缺血具有强大的保护作用后，我们将在此基础上进一步提高心肌细胞的修复能力。在目标1中，我们将确定HO-1和CO在调节CPC功能中的作用。在Aim 2中，我们将建立ecSOD在调节CPC功能和介导HO-1诱导的CPC保护中的作用。在Aim 3中，我们将确定NO在调节CPC功能中的作用，以及HO-1和ecSOD在介导NO对CPC的影响中的作用。在这三个目的中，我们将使用功能获得和功能丧失的方法，系统地评估体外CPC能力的基本参数，以及心肌梗死后左室重构和功能障碍小鼠模型中CPC在体内修复心脏损伤的能力。在目标4。我们将阐明CO和NO上调ecSOD的分子机制，以及NO在cpc中上调HO-1和ecSOD的分子机制，重点研究转录因子Nrf2。这将是首次研究CO和NO及其支持蛋白HO-1、ecSOD和INOS在调节CPC功能中的作用。结果将是全新的，并将为我们对CPC生物学的理解增加一个新的维度。此外，这些研究将为增强CPCs中CO-NO模块的操作的治疗效用提供原理证明，这可能为未来在心衰患者中进行基因或药理学增强的CPCs试验奠定基础。

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 transplantation. This implies that increasing the survival of transplanted cells by preventing 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 Project 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 dramatically 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.