ERECTILE DYSFUNCTION AND NITRIC OXIDE SYNTHASE IN AGING

衰老过程中的勃起功能障碍和一氧化氮合酶

• 批准号: 6619955
• 负责人: Nestor F Gonzalez-Cadavid
• 金额: $ 22.45万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6619955
• 关键词: NMDA receptors; aging; apoptosis; complementary DNA; confocal scanning microscopy; gene expression; gene targeting; gene therapy; genetically modified animals; geriatrics; immunocytochemistry; laboratory mouse; laboratory rat; muscle cells; nitric oxide synthase; penis disorder; penis erection; polymerase chain reaction; smooth muscle; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Erectile dysfunction (ED) affects over 15 million men and their partners in the USA. Aging is a major risk factor for ED, presumably due to an insufficient synthesis of nitric oxide (NO) in the penile nerves and an impaired compliance of the corpora cavernosa (CC) caused by a loss of smooth muscle cells (SMC). Failure to respond to PDE5 inhibitors may be related to these factors. The overall .goal of this proposal is to extend our previous grant findings towards a curative approach for aging-related ED based on: a) increasing the content and activity of penile neuronal nitric oxide synthase (PnNOS) in the CC, to release more NO upon sexual stimulation; and b) decreasing the levels of inducible NOS (iNOS) and reactive oxygen species (ROS) in the CC to reduce peroxynitrite formation and SMC apoptosis. We propose: Aim 1: to improve the duration of intracavernosal gene therapy with PnNOS cDNA, by: a) stimulating persistence of gene expression in the penis of adult rats using novel helper-dependent (HD) adenovirus (AdV) and tissue-specific promoters directing the expression of beta-galactosidase; b) determining whether hybrid AdV/adeno-associated virus (AdV-AAV) are more effective than HD-AdV; c) comparing electroporation and minipumps for construct delivery, and conducting safety tests, and d) applying the selected vectors or cells to express PnNOS and related genes in the aged CC, to correct the impaired erectile response to electrical stimulation of the cavernosal nerve; Aim 2: to characterize modulators of PnNOS activity that may affect the control of erection in the CC, by: a) stimulating PnNOS activity and erectile function through the activation of nNOS-associated NMDA receptors; and b) finding novel PnNOS binding protein factors modulating its activity and erectile function; Aim 3: to correct the aging-related loss of SMC by manipulating the NO/ROS ratio within the CC, by a) determining whether there is SMC replication and whether this is decreased with aging, and correlating with the increase in iNOS, ROS, peroxynitrite, and apoptosis; b) reducing NO (from iNOS) and ROS levels to reverse the loss of SMC and ameliorate aging-related ED; and c) determining whether obliteration of iNOS expression in the iNOS knock-out mouse leads to a reduction in the age-related loss of SMC.

描述（由申请人提供）：勃起功能障碍（ED）影响了1500万人及其在美国的伴侣。衰老是ED的主要危险因素，可能是由于阴茎神经中一氧化氮（NO）的合成不足，并且由于流失肌肉细胞（SMC）而导致的Corpora Cavernosa（CC）的依从性受损。对PDE5抑制剂的反应可能与这些因素有关。该提案的总体目标是将我们先前的赠款发现扩展到基于衰老相关的ED的治疗方法：性刺激； b）降低CC中可诱导的NOS（INOS）和活性氧（ROS）的水平，以减少过氧亚硝酸盐的形成和SMC凋亡。我们提出：目标1：用PNNOS cDNA改善腹腔内综合基因治疗的持续时间，作者：指导β-半乳糖苷酶的表达； b）确定混合ADV/腺相关病毒（ADV-AAV）是否比HD-ADV更有效； c）比较电穿孔和微型固定，用于构造和进行安全测试，d）在老年CC中应用选定的载体或细胞表达PNNOS和相关基因，以纠正勃起对海绵体膜神经电刺激的勃起反应受损；目的2：表征可能影响CC中勃起的PNNOS活性调节剂，通过：a）通过激活NNOS相关的NMDA受体来刺激PNNOS活性和勃起功能； b）找到新型的PNNOS结合蛋白因子调节其活性和勃起功能；目标3：通过操纵CC内的NO/ROS比率来纠正SMC的衰老相关损失，a）确定是否存在SMC复制，以及与衰老相关，并与Inos，ROS，ROS，ROS，ROS，ROS，ROS，ROS，ROS，ROS，相关过氧亚硝酸盐和凋亡； b）减少NO（从INOS）和ROS水平降低SMC的损失并改善与衰老相关的ED； c）确定iNOS敲除小鼠中iNOS表达的闭塞是否会导致与年龄相关的SMC损失的减少。