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: 6564793
• 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: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564793
• 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出血后常见的毁灭性并发症， 出血（SAH），各种治疗方法都失败了。 有证据表明降钙素基因相关肽 SAH后三叉神经感觉神经中的降钙素基因相关肽（CGRP）耗尽，这意味着 一个重要的补偿机制可能会用尽。相比 对其他几种脑血管扩张剂的反应减弱， 对CGRP的反应在SAH后保持或增强。 这项研究的一个目标是确定基因是否 体外移植可改变SAH后脑血管功能。的 研究者已经制备了重组腺病毒载体， 内皮型一氧化氮合酶（eNOS）和前降钙素基因相关肽原（prepro-CGRP）。研究是 建议确定是否在体外基因转移eNOS，诱导 NOS（iNOS）和prepro-CGRP产生颅内动脉舒张 SAH后。调查人员将使用他们开发的方法， 研究基因转移到体外血管中。 第二个目标是检查SAH对转基因表达的影响。的 研究者已经观察到β半乳糖苷酶的显著表达， 在基因转移到SAH后的狗后，使用具有 巨细胞病毒（CMV）启动子驱动β半乳糖苷酶的表达。 建议进行研究以确定转基因表达是否是 当腺病毒载体与CMV，但不是Rous 肉瘤病毒（RSV），使用启动子，可能是因为应答元件 在CMV启动子中的NrkappaB被转移到细胞核中激活 对氧化应激的反应。这些实验应该阐明 导致SAH后转基因表达增强。 第三个目标是确定CGRP的基因转移是否可以改变 脑血管功能和预防SAH后血管痉挛。 提出研究以确定是否基因转移eNOS，iNOS， 脑脊髓液注射腺病毒载体制备降钙素基因相关肽前体 抑制SAH后血管痉挛的发展。研究人员计划 使用他们开发的基因转移到大脑的方法， 血管和血管周围组织。血管反应将是 在大鼠和犬模型中检查，以及对犬血管痉挛的影响 模型将被确定。即使有目前可用的载体， 提供延迟和瞬时转染，基因治疗，以防止 SAH后可能出现血管痉挛。