Role of the NO-CO Module in Regulating CPC Function

NO-CO 模块在调节 CPC 功能中的作用

• 批准号: 8492142
• 负责人: Roberto Bolli
• 金额: $ 30.8万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8492142
• 关键词: Apoptosis; Apoptotic; Carbon Monoxide; Cardiac; Cell Survival; Cell Therapy; Cell Transplants; Cell physiology; Cells; Cellular biology; Chemotaxis; Clinical Research; Competence; Deet; Dimensions; Effectiveness; Evolution; Functional disorder; Funding; Future; Gene Transfer; Generations; Genetic; Genetic Transcription; Goals; Heart failure; Heme; Human; In Vitro; Instruction; Ischemia; Left Ventricular Remodeling; Length; Measures; Mediating; Mediator of activation protein; Minority; Modeling; Molecular; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase; Oxidative Stress; Patients; Phase I Clinical Trials; Proteins; Proto-Oncogene Protein c-kit; Reactive Oxygen Species; Resistance; Role; Stem cells; Structure; Superoxide Dismutase; Telomerase; Testing; Therapeutic; Translations; Transplantation; Up-Regulation; Work; cardiac repair; clinical application; cytokine; extracellular; gain of function; heme oxygenase-1; human NOS2A protein; in vivo; inhibitor/antagonist; loss of function; novel strategies; oxidative damage; phase 1 study; pre-clinical; preconditioning; prevent; programs; senescence; telomere; transcription factor

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.

心脏前体细胞(CPC)的发现为治疗心力衰竭(HF)提供了一种潜在的新方法，我们对心衰患者(Scipio)中的CPC进行的首次人体临床研究的初步结果令人鼓舞。然而，基于CPC的治疗受到以下事实的严重限制：几乎所有(至少97%)移植的CPC在移植后不久死亡。这意味着，通过阻止细胞凋亡来提高移植细胞的存活率将增强CPC治疗的疗效。在本项目目前的资助期间，我们发现一氧化碳(CO)和一氧化氮(NO)具有强大的抗细胞凋亡作用，并形成一个密切相关的功能模块(CO-NO模块)，该模块受血红素加氧酶-1(HO-1)、细胞外超氧化物歧化酶(EcSOD)和诱导型一氧化氮合酶(INOS)的调节。我们将利用这一发现来加强基于细胞的疗法。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后的治疗作用。我们认为，增强这个模块将极大地增强移植的CPC的有效性，并显著增强CPC介导的心脏修复。这个项目代表了我们之前在这个项目中的工作的自然发展；在发现CO-NO模块对心肌缺血提供了强大的保护后，我们现在将在这一工作的基础上加强CPC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。在目标2中，我们将确定EcSOD在调节CPC功能和介导HO-1诱导的CPC保护中的作用。在目标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CCs影响中的作用。在这三个目标中，我们将使用获得和丧失功能的方法，系统地评估体外CPC能力的基本参数，以及在心肌梗死后左室重构和功能障碍的小鼠模型中CPC修复心脏损伤的能力。在目标4中，我们将以转录因子Nrf2为重点，阐明CO和NO上调CCs中EcSOD的分子机制，以及NO上调HO-1和EcSOD的分子机制。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。这些结果将是全新的，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CPC中CO-NO模块的操作的治疗效用提供原则证明，这可能为未来在心力衰竭患者中进行遗传或药物增强的CPC的试验奠定基础。