A Model for Vasopressin Gene Construct Expression In Vivo

加压素基因构建体内表达模型

• 批准号: 9311307
• 负责人: Thomas Sherman
• 金额: $ 28.57万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9311307&HistoricalAwards=false
• 关键词: Model; Vasopressin; Gene; Construct; Expression

9311307 Sherman Vasopressin is a peptide hormone that is synthesized in magnocellular neurons of the hypothalamus and is secreted into the blood stream from nerve endings in the posterior pituitary. The primary stimulus for vasopressin release by the posterior pituitary is dehydration. Indeed, this neuropeptide is the antidiuretic hormone. It is essential for the regulation of water balance. In addition, vasopressin is involved in stress mechanisms since it acts synergistically with the hypothalamic- pituitary-adrenal axis to stimulate secretion of adrenocorticotropic hormone. Much of our knowledge about vasopressin has resulted from work with the Brattleboro rat. These rats do not synthesize vasopressin because of a single base deletion in the gene which is inherited as an autosomal recessive trait. As a consequence of this gene defect, Brattleboro rats develop diabetes insipidus. Dr. Sherman is examining the use of a modified herpes virus to facilitate in vivo investigations of vasopressin structure- function relationships. The herpes virus type 1 (HSV-1) is uniquely capable of infecting non-dividing cells, such as neurons. Moreover, it has the added characteristic of existing in neurons, under some conditions, in a dormant or quiescent state termed latency, during which only a small portion of the herpes genome is active. Using newly developed state-of-the-art techniques, Dr. Sherman will engineer a special modified HSV-1 virus to incorporate a working version of the gene for vasopressin. He will develop different versions of normal vasopressin gene constructions and various HSV-1 viral modifications to maximize the expression of normal vasopressin hormone in Br attleboro rats. Results from this work are likely to make significant contributions both within the field of vasopressin research and the development and use of HSV vectors for gene expression. Indeed, the ultimate goal is to correct the diabetes insipidus defect by specifically targeting magnocellular neurons by direct injections of the vasopressin-containing virus into the posterior pituitary, where the virus particles can be taken up by the magnocellular nerve terminals, retrogradely transported to the hypothalamic cell bodies, establish latency, and begin expressing the virally-contained vasopressin gene. This work provides an important bridge between neuroscience and virology and leads the way for new developments in gene therapy. ***

9311307谢尔曼加压素是一种肽激素，在下丘脑的大细胞神经元中合成，并从垂体后叶的神经末梢分泌到血流中。 垂体后叶释放加压素的主要刺激是脱水。 事实上，这种神经肽是抗利尿激素。 它对水平衡的调节至关重要。 此外，加压素参与应激机制，因为它与下丘脑-垂体-肾上腺轴协同作用以刺激促肾上腺皮质激素的分泌。 我们对后叶加压素的了解大部分来自于对布拉特伯勒大鼠的研究。 这些大鼠不合成加压素，因为该基因中的一个碱基缺失是作为常染色体隐性遗传性状遗传的。 由于这种基因缺陷，Brattleboro大鼠发生尿崩症。 谢尔曼博士正在研究使用一种改良的疱疹病毒来促进加压素结构-功能关系的体内研究。疱疹病毒1型（HSV-1）是唯一能够感染非分裂细胞，如神经元。 此外，它还具有存在于神经元中的额外特征，在某些条件下，处于休眠或静止状态，称为潜伏期，在此期间，只有一小部分疱疹基因组是活跃的。 利用最新开发的最先进的技术，谢尔曼博士将设计一种特殊的改良HSV-1病毒，将加压素基因的工作版本。 他将开发不同版本的正常加压素基因结构和各种HSV-1病毒修饰，以最大限度地提高正常加压素激素在布拉特伯勒大鼠中的表达。 这项工作的结果可能会在加压素研究领域以及HSV基因表达载体的开发和使用方面做出重大贡献。 实际上，最终目标是通过将含加压素的病毒直接注射到垂体后叶中，特异性靶向大细胞神经元来纠正尿崩症缺陷，在垂体后叶中，病毒颗粒可以被大细胞神经末梢摄取，逆行转运到下丘脑细胞体，建立潜伏期，并开始表达含病毒的加压素基因。 这项工作提供了神经科学和病毒学之间的重要桥梁，并为基因治疗的新发展铺平了道路。 ***