NOS GENE TRANSFER IN PULMONARY HYPERTENSION

这项建议的总体目的是审查以下各项之间的平衡 一氧化氮(NO)的产生和超氧化物介导的天然气破坏 正常和低氧/高血压的肺循环，努力 建立肺组织中基因转移的目的基因 高血压(PHTN)。NO是一种重要的抑制性调节因子 肺动脉张力和中膜平滑肌细胞增殖。 在PHTN的发生发展过程中，内源性NO活性不足 完全反对血管收缩和血管壁重塑。 这种相对不足的一氧化氮活动尽管增加了 内皮型一氧化氮合酶(ENOS)的表达 提示反调节因子可能抑制内源性eNOS 或加速销毁NO。重要的机制，通过它 NO可通过与超氧阴离子反应而被破坏。这种反应 可被超氧化物歧化酶(SOD)预防。我们假设 在PHTN的发展过程中，NOS和SOD活性不足可能 单独或组合导致次最大生物可利用NO， 导致肺血管收缩和血管壁重塑。 因此，我们建议检验这样一种假设，即肺活量减少 一氧化氮合酶或超氧化物歧化酶活性增强小鼠肺血管收缩 通过靶向删除eNOS、诱导型(I)NOS、细胞外 (EC-SOD，或铜，锌-超氧化物歧化酶。我们还将测试反向假设，即 肺组织NO生成量增加，肺血管收缩 在体内，通过瞬时过表达一氧化氮合酶(eNOS或eNOS)而减弱 INOS)和超氧化物歧化酶(EC-SOD或铜、锌-超氧化物歧化酶)。表达式将为 肺亲和性阳离子注射靶向肺内皮细胞 脂质/DNA复合体。使用淘汰赛和淘汰赛相结合的方法 瞬时过表达感兴趣的基因，我们会更多 明确确定一氧化氮合酶和超氧化物歧化酶在调节细胞周期中的作用 PHTN的开发比以前使用的可能 药理学方法。虽然我们的重点包括详细的 一氧化氮合酶和超氧化物歧化酶调节作用的生理学评价 肺血管张力，我们的总体目标是开发新的 对患有PHTN的人类的治疗。因此，检测NOS和/或SOD 过度表达减轻肺血管收缩将有助于 找出PHTN“基因治疗”的潜在基因，如 并提供肺血管基因原理的证据 作为一种新的治疗方法，移植可能值得进一步研究。 PHTN.

The overall purpose of this proposal is to examine the balance between nitric oxide (NO) production and superoxide-mediated NG destruction in the normal and hypoxic/hypertensive pulmonary circulation, in an effort to establish genes of interest for gene transfer in pulmonary hypertension (PHTN). NO is an important inhibitory modulator of pulmonary artery tone and medial smooth muscle cell proliferation. During the development of PHTN, endogenous NO activity is insufficient to completely oppose vasoconstriction and vascular wall remodeling. This relative insufficiency of NO activity occurs despite increased expression of endothelial constitutive nitric oxide synthase (eNOS), suggesting that counter regulatory factors may inhibit endogenous eNOS or accelerate destruction of NO. An important mechanism through which NO may be destroyed is reaction with superoxide anion. This reaction can be prevented by superoxide dismutase (SOD). We hypothesize that during the development of PHTN, insufficient NOS and SOD activity may independently or in combination lead to submaximal bioavailable NO, resulting in pulmonary vasoconstriction and vascular wall remodeling. We therefore propose to test the hypothesis that reduction in pulmonary NOS or SOD activity will enhance pulmonary vasoconstriction using mice with targeted deletion of either eNOS, inducible (i)NOS, extracellular (EC-SOD, or Cu,Zn-SOD. We will also test the converse hypothesis, that pulmonary NO production can be augmented, and pulmonary vasoconstriction attenuated, in vivo, by transient overexpression of NOS (either eNOS or iNOS), and SOD (either EC-SOD or Cu,Zn-SOD). Expression will be targeted to pulmonary endothelium by injection of lung-avid cationic lipid/DNA complexes. Using the combined approaches of knockout and transient overexpression of the genes of interest, we will more definitively establish the roles of NOS and SOD in modulating the development of PHTN than has previously been possible using pharmacological approaches. While our focus includes a detailed physiological evaluation of the roles of NOS and SOD in modulating pulmonary vascular tone, our overall goal is development of novel therapies for humans with PHTN. Thus, testing if NOS and/or SOD overexpression attenuates pulmonary vasoconstriction will serve both to identify potential genes of interest for "gene therapy" for PHTN, as well as providing proof of principle that pulmonary vascular gene transfer could merit future study as a novel therapeutic approach in PHTN.