Gene Therapy in the Senile Brain and Hypophysis

老年大脑和垂体的基因治疗

• 批准号: 7122720
• 负责人: RODOLFO G GOYA
• 金额: $ 4.08万
• 依托单位: NATIONAL UNIVERSITY OF LA PLATA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7122720
• 关键词: Adenoviridae; South America; aging; animal old age; cell death; dopamine; female; gene delivery system; gene expression; gene therapy; hypothalamus; immunocytochemistry; international cooperation; laboratory rat; neoplasm /cancer chemotherapy; nervous system disorder therapy; neural degeneration; neurons; neuroprotectants; neurotrophic factors; nonhuman therapy evaluation; pituitary gland; pituitary neoplasms; prolactin; reporter genes; stereotaxic techniques; technology /technique development; transfection /expression vector

DESCRIPTION (provided by applicant): The increase of the elderly population in the urban areas of Argentina is comparable to that of many North American and European cities. Consequently, the incidence of age-related neurological pathologies is becoming a problem of significant medical and economic impact for the country. In this context, the overall goal of the present proposal is to establish a long-term collaboration between the American and Argentine applicants, who share an interest in the potential of gene therapy for the treatment of neurodegenerative diseases. Three specific objectives are proposed: 1) To assess the performance of self-regulating adeno associated viral (AAV) vectors in the brain of old rats. These vectors harbor a reporter gene encoding humanized green fluorescent protein (hGFP) under the control of the tet-off or tet-on promoter system which can be turned off or on, respectively by administration of tetracycline (Tet) or certain Tet derivatives. The above vectors will be stereotaxically injected in the paraventricular, periventricular and arcuate nuclei, all of which contain dopaminergic (DA) neurons of known vulnerability to aging. 2) To assess the therapeutic efficacy of a self-regulating tet-off AAV vector harboring the gene for human glial cell line-derived neurotrophic factor (hGDNF), a potent neuroprotective molecule. The vector will be stereotaxically injected in the arcuate nucleus of 24-month old rats and 6 months later, its ability to prevent the normal age-related loss of hypothalamic tuberoinfundibular DA (TIDA) neurons will be assessed by quantitative immuno-histochemistry for DA neurons and ELISA for hGDNF. Serum PRL levels (an index of TIDA neuron function) will be monitored in the animals throughout the experiment. 3) To construct two self-regulating adenoviral vectors (RAd) harboring suicide genes under the control of the tet-off system. One of the RAds will harbor a hybrid suicide gene coding for a fusion protein between hGFP and HSV1 thymidine kinase (HSV1-TK), which retains fluorescence and HSV1-TK suicide activity. The other RAd will harbor the separate genes for HSV1-TK and hGFP under the control of a bi-directional Tet-off regulatory element. These two vectors, which are expected to be safer than non-regulatable counterparts, will be employed to treat experimental rat prolactinomas. Further to the above specific objectives, contacts with basic and clinical neuroscientists are planned in the US and Argentina in order to assemble a follow up research program focused on senile brain disorders.

描述（由申请人提供）：阿根廷城市地区老年人口的增长与许多北美和欧洲城市相当。 因此，与年龄相关的神经系统疾病的发病率正在成为对该国产生重大医疗和经济影响的问题。 在这种背景下，本提案的总体目标是在美国和阿根廷申请人之间建立长期合作，他们对基因疗法治疗神经退行性疾病的潜力有着共同的兴趣。 提出了三个具体目标：1）评估老年大鼠大脑中自我调节腺相关病毒（AAV）载体的性能。 这些载体含有编码人源化绿色荧光蛋白（hGFP）的报告基因，该报告基因受tet-off或tet-on启动子系统的控制，可以通过施用四环素（Tet）或某些Tet衍生物分别关闭或打开。 上述载体将立体定向注射到室旁核、室周核和弓状核中，所有这些核都含有已知易受衰老影响的多巴胺能 (DA) 神经元。 2) 评估带有人胶质细胞源性神经营养因子（hGDNF）（一种有效的神经保护分子）基因的自我调节 tet-off AAV 载体的治疗效果。 该载体将被立体定位注射到 24 个月大的大鼠的弓状核中，6 个月后，将通过 DA 神经元的定量免疫组织化学评估其防止下丘脑结节漏斗部 DA (TIDA) 神经元正常年龄相关损失的能力以及 hGDNF 的 ELISA。 在整个实验过程中将监测动物的血清 PRL 水平（TIDA 神经元功能指数）。 3)构建两个在tet-off系统控制下携带自杀基因的自我调节腺病毒载体(RAd)。 其中一个 RAd 将包含一个混合自杀基因，编码 hGFP 和 HSV1 胸苷激酶 (HSV1-TK) 之间的融合蛋白，该融合蛋白保留荧光和 HSV1-TK 自杀活性。 另一个 RAd 将在双向 Tet-off 调控元件的控制下包含 HSV1-TK 和 hGFP 的独立基因。 这两种载体预计比不可调节的载体更安全，将用于治疗实验性大鼠泌乳素瘤。 为了实现上述具体目标，计划与美国和阿根廷的基础和临床神经科学家进行接触，以制定针对老年脑部疾病的后续研究计划。

项目成果

Fatty acid profiles in hepatic membranes of rats with different levels of circulating estrogen and prolactin.

循环雌激素和催乳素水平不同的大鼠肝膜中的脂肪酸谱。

• 发表时间: 2007-04
• 作者: Bellini, Maria J;Carino, Monica H;Tacconi;Goya, Rodolfo G
• 通讯作者: Goya, Rodolfo G

Peripheral and mesencephalic transfer of a synthetic gene for the thymic peptide thymulin.

胸腺肽胸腺素合成基因的外周和中脑转移。

• 发表时间: 2006-05-31
• 作者: Morel, Gustavo R;Brown, Oscar A;Reggiani, Paula C;Herenu, Claudia B;Portiansky, Enrique L;Zuccolilli, Gustavo O;Pleau, Jean M;Dardenne, Mireille;Goya, Rodolfo G
• 通讯作者: Goya, Rodolfo G

Dopaminergic mesencephalic systems and behavioral performance in very old rats.

高龄大鼠的多巴胺能中脑系统和行为表现。

• 发表时间: 2008-07-17
• 影响因子: 3.3
• 作者: Sanchez, H L;Silva, L B;Portiansky, E L;Herenu, C B;Goya, R G;Zuccolilli, G O
• 通讯作者: Zuccolilli, G O

Gene therapy in the neuroendocrine system.

神经内分泌系统的基因治疗。

• 发表时间: 2006
• 作者: Hereñú, C B;Morel, G R;Bellini, M J;Reggiani, P C;Sosa, Y E;Brown, O A;Goya, R G
• 通讯作者: Goya, R G

The thymus-neuroendocrine axis: physiology, molecular biology, and therapeutic potential of the thymic peptide thymulin.

胸腺-神经内分泌轴：胸腺肽胸腺素的生理学、分子生物学和治疗潜力。

• 发表时间: 2009-02
• 影响因子: 5.2
• 作者: Reggiani PC;Morel GR;Cónsole GM;Barbeito CG;Rodriguez SS;Brown OA;Bellini MJ;Pléau JM;Dardenne M;Goya RG
• 通讯作者: Goya RG