MODULATION OF NO WITH PULMONARY GENE TRANSFER

通过肺部基因转移调节 NO

• 批准号: 6725425
• 负责人: Loretta G Que
• 金额: $ 12.49万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6725425
• 关键词: apoptosis; arginase; biotechnology; cell proliferation; chemokine; enzyme activity; free radical oxygen; gene therapy; histology; hyperoxia; immunocytochemistry; inflammation; laboratory rabbit; lung injury; nitric oxide; nitric oxide synthase; oxidative stress; superoxide dismutase; transfection; transfection /expression vector; western blottings

Acute lung injury in the intensive care unit is treated with administration of high inspired oxygen tensions which can be toxic to the lung. Pulmonary oxygen toxicity is generally attributed to the excessive production of reactive oxygen intermediates (RO1), e.g., superoxide (O2), hydrogen peroxide (H2O2), and hydroxyl radicals. Recently nitric oxide (NO) is being used to treat hyperoxic lung disease, but its effect is variable and its role remains controversial. We hypothesize that selective modulation of NO/RO1 in different cellular compartments regulates tissue responses to hyperoxia. In this proposal, we will employ state of the art adenoviral gene transfer technique in vitro and in vivo. The transgenes we have selected for these studies are neuronal nitric oxide synthase (nNOS), extracellular superoxide dismutase (ECSOD), and arginases I and II (AI and AII). Overexpression of these four genes are expected to affect NO and reactive oxygen intermediates (RO1) production in different cell compartments: nNOS (yields intracellular NO/yields 02-), ECSOD (yields extracellular NO/yields 02-), and AI and AII (yields intracellular NO/yields O2-). We will determine how these transgenes modulate, oxidative/nitrosative stress, and NO/ROI mediated inflammation, cellular proliferation, and apoptosis. The specific aims of this proposal are 1) characterize the transgene expression of nNOS, ECSOD, AI, and AII in vitro 2) characterize intratracheal adenoviral gene transfer in normal rabbit lung and 3) evaluate how adenoviral mediated transgenes nNOS, ECSOD, AI, and AII modulate effect of NO in hyperoxia. These studies should not only better unravedl the basic mechanisms of hyperoxic lung injury, but also provide a rationale basis for therapeutic gene transfer using NO. Ultimately we are hopeful that these results may also extend to other oxidant injuries in the lung.

重症监护病房的急性肺损伤是通过高吸入氧张力来治疗的，高吸入氧张力可能对肺部有毒。 肺氧毒性通常归因于活性氧中间体（RO1）的过量产生，例如超氧化物（O2）、过氧化氢（H2O2）和羟基自由基。 最近，一氧化氮（NO）被用于治疗高氧性肺病，但其效果参差不齐，其作用仍存在争议。 我们假设不同细胞区室中 NO/RO1 的选择性调节可调节组织对高氧的反应。在本提案中，我们将在体外和体内采用最先进的腺病毒基因转移技术。 我们为这些研究选择的转基因是神经元一氧化氮合酶 (nNOS)、细胞外超氧化物歧化酶 (ECSOD) 以及精氨酸酶 I 和 II（AI 和 AII）。 这四个基因的过度表达预计会影响不同细胞区室中 NO 和活性氧中间体 (RO1) 的产生：nNOS（产生细胞内 NO/产生 02-）、ECSOD（产生细胞外 NO/产生 02-）以及 AI 和 AII（产生细胞内 NO/产生 O2-)。 我们将确定这些转基因如何调节氧化/亚硝化应激以及 NO/ROI 介导的炎症、细胞增殖和细胞凋亡。 该提案的具体目标是 1) 表征体外 nNOS、ECSOD、AI 和 AII 的转基因表达 2) 表征正常兔肺中气管内腺病毒基因转移 3) 评估腺病毒如何介导转基因 nNOS、ECSOD、AI 和 AII AII 在高氧条件下调节 NO 的作用。这些研究不仅可以更好地揭示高氧性肺损伤的基本机制，而且可以为利用NO进行治疗性基因转移提供理论基础。最终，我们希望这些结果也能扩展到肺部的其他氧化损伤。

项目成果

Protection from experimental asthma by an endogenous bronchodilator.

通过内源性支气管扩张剂预防实验性哮喘。

• 发表时间: 2005-06-10
• 作者: Que, Loretta G;Liu, Limin;Yan, Yun;Whitehead, Gregory S;Gavett, Stephen H;Schwartz, David A;Stamler, Jonathan S
• 通讯作者: Stamler, Jonathan S