CALCITONIN GENE REGULATED PEPTIDE IN SUBARACHNOID HEMORRHAGE--GENE THERAPY

降钙素基因调控肽在蛛网膜下腔出血中的作用--基因治疗

• 批准号: 6415220
• 负责人: DONALD D HEISTAD
• 金额: $ 23.33万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6415220
• 关键词: angiotensin II; calcitonin gene related peptide; cerebral hemorrhage; dogs; gene expression; gene therapy; genetic promoter element; hemodynamics; laboratory rat; neuroprotectants; nitric oxide synthase; nonhuman therapy evaluation; subarachnoid space; transfection /expression vector; trigeminal nerve; vasoconstrictors; vasomotion; vasospasm

Vasospasm is a common and devastating complication after subarachnoid hemorrhage (SAH), and a variety of therapeutic approaches have failed. Several lines of evidence indicate that calcitonin gene-related peptide (CGRP) is depleted from trigeminal sensory nerves after SAH, which implies that an important compensatory mechanism may be exhausted. In contrast to diminished responses to several other cerebral vasodilators, vasodilator responses to CGRP are preserved or enhanced after SAH. One goal of the studies that are proposed is to determine whether gene transfer in vitro can alter cerebral vascular function after SAH. The investigators have prepared recombinant adenoviral vectors that express endothelial nitric oxide synthase (eNOS) and prepro-CGRP. Studies are proposed to determine whether gene transfer in vitro of eNOS, inducible NOS (iNOS), and prepro-CGRP produces relaxation of intracranial arteries after SAH. The investigators will use a method that they have developed to study gene transfer to blood vessels in vitro. A second goal is to examine effects of SAH on transgene expression. The investigators have observed pronounced expression of beta galactosidase after gene transfer to dogs after SAH, using an adenoviral vector with a cytomegalovirus (CMV) promoter driving expression of beta galactosidase. Studies are proposed to determine whether transgene expression is increased by SAH when an adenoviral vector with a CMV, but not Rous sarcoma virus (RSV), promoter is used, perhaps because response elements in the CMV promoter are activated by NrkappaB translocated to the nucleus in response to oxidant stress. These experiments should clarify mechanisms that lead to augmented transgene expression after SAH. A third goal is to determine whether gene transfer of CGRP can alter cerebral vascular function and prevent vasospasm in vivo after SAH. Studies are proposed to determine whether gene transfer of eNOS, iNOS, and prepro-CGRP by injection of adenoviral vectors into cerebrospinal fluid inhibits development of vasospasm following SAH. The investigators plan to use a method that they have developed for gene transfer to cerebral vessels and perivascular tissue in vivo. Vascular responses will be examined in rat and canine models, and effects on vasospasm in canine model will be determined. Even with currently available vectors, which provide delayed and transient transfection, gene therapy to prevent vasospasm after SAH may be possible.

血管痉挛是蛛网膜下腔术后常见且严重的并发症。 出血(SAH)，以及各种治疗方法都失败了。 多条证据表明降钙素基因相关肽 SAH后三叉神经降钙素基因相关肽(CGRP)耗竭，提示 一个重要的补偿机制可能会耗尽。与之形成鲜明对比的是 对其他几种脑血管扩张剂、血管扩张剂的反应减弱 SAH后对CGRP的反应被保留或增强。 被提议的研究的一个目标是确定基因 体外移植可改变蛛网膜下腔出血后的脑血管功能。这个 研究人员已经制备了重组腺病毒载体，可以表达 内皮型一氧化氮合酶(ENOS)和降钙素基因相关肽(CGRP)前体的表达。研究是 建议确定eNOS的体外基因转移是否可诱导 一氧化氮合酶(INOS)和降钙素基因相关肽前体对颅内动脉的松弛作用 在SAH之后。调查人员将使用他们开发的一种方法 体外血管基因转移的研究。 第二个目标是检测SAH对转基因表达的影响。这个 研究人员观察到β-半乳糖苷酶的显著表达 在SAH后将基因转移到狗身上后，使用腺病毒载体 巨细胞病毒(CMV)启动子驱动β-半乳糖苷酶表达。 有人建议进行研究以确定转基因表达是否 当腺病毒载体携带CMV而不是ROU时，SAH增加 使用肉瘤病毒(RSV)启动子，可能是因为反应元件 在CMV启动子中被移位到细胞核的NrkappaB激活 以应对氧化应激。这些实验应该能弄清机制 这导致SAH后转基因表达增强。 第三个目标是确定CGRP的基因转移是否会改变 SAH后体内脑血管功能及预防血管痉挛。 有人建议进行研究，以确定eNOS、iNOS和 脑脊液注射腺病毒载体前降钙素基因相关肽 抑制SAH后血管痉挛的发展。调查人员计划 使用他们开发的将基因转移到大脑的方法 体内的血管和血管周围组织。血管反应将是 在大鼠和犬模型上进行检测，并对犬血管痉挛的影响 型号将会确定。即使使用当前可用的矢量，也可以 提供延迟和瞬时转染法，防止基因治疗 蛛网膜下腔出血后血管痉挛是可能的。