GENETIC REGULATION OF THE HEPATIC ACUTE PHASE RESPONSE

肝脏急性期反应的基因调控

• 批准号: 3232301
• 负责人: HEINZ BAUMANN
• 金额: $ 16.14万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1992-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3232301
• 关键词: alpha 1 antitrypsin; animal population genetics; biological models; complementary DNA; gene expression; genetic library; genetic mapping; genetic regulation; genetic transcription; genetically modified animals; glucocorticoids; hemopexin; hemostasis; hormone regulation /control mechanism; kidney function; laboratory mouse; liver cells; molecular cloning; molecular genetics; nucleic acid hybridization; orphan disease /drug; plasma cells; tissue /cell culture

Homeostasis of mammals is strongly dependent on correct developmental stage-, tissue- and hormone-specific expression of plasma proteins. Regulated plasma protein production is especially important during systemic injury when the body's demand for essential plasma proteins must be met by an immediate increase in hepatic synthesis. The overall goal of this project is to characterize the largely unknown genetic elements governing the activity of plasma protein genes, in particular those induced by systemic injury. In order to identify such elements, inbred strains and wild derived species of mice had been surveyed for genetic variants in plasma protein gene expression. M. caroli revealed two exceptional features: 1) the structural gene for alpha 1-acid glycoprotein (AGP) has been amplifed to 4-6 copies, each encoding distinct forms with different degrees of inducibility by inflammation, and 2) the regulation of the single, liver specific gene for alpha1-antitrypsin (AT) has been altered such that the gene is equally expressed in liver and in kidney. These unique genetic properties, evolved in M. caroli, will be used to assess those gene elements determining a) the acute phase induction of AGP gene and b) the tissue-specificity of the AT gene expression. The experimental approaches will be as follows: 1) The structural genes for high and low inducible AGPs and for AT will be isolated. 2) The transcriptional activity and regulatory response of the different AGP genes will be tested by introducing the genes into human and rat hepatoma cells which are responsive to the two most potent mediators of hepatic acute phase response, the hepatocyte stimulating factor and glucocorticoids. To identify the liver and kidney specificity of AT gene expression, the activity of the M. caroli AT gene will be measured and compared to the M. domesticus AT gene by transfection into hepatic and renal tissue culture cells. After demonstrating correct regulation of the AGP and AT genes, the cis-acting regulatory elements will be localized relative to the structural genes. 3) The extent to which functional AGP and AT gene sequences support correct tissue, hormone and developmental regulation will be assessed in transgenic mice. 4) Lastly, the cellular factors interacting specifically with the cis-acting regulatory elements will be identified. The results from these studies on M. caroli genes will provide new insights into the signals controlling tissue specific and inflammation-inducibility of plasma protein genes.

哺乳动物的体内平衡强烈依赖于正确的 发育阶段、组织和胚胎特异性表达 血浆蛋白 调节血浆蛋白的产生是 尤其是在全身性损伤时， 对必需血浆蛋白的需求必须通过 肝脏合成立即增加。 总的目标是 该项目旨在描述大部分未知的遗传因素， 控制血浆蛋白基因的活性，特别是那些 由全身性损伤引起。 为了确定这些要素， 调查了小鼠的近交系和野生衍生种 血浆蛋白基因表达的遗传变异。 M. Caroli 揭示了两个特殊的特征：1）结构基因， α 1-酸性糖蛋白（AGP）已扩增至4 - 6个拷贝， 每个编码具有不同程度的诱导的不同形式 通过炎症，和2）单一的，肝脏特异性的调节 α 1-抗胰蛋白酶（AT）基因已被改变， 该基因在肝脏和肾脏中同等表达。 这些独特 遗传特性，在M. Caroli将用于评估 这些基因元件决定a）急性期诱导 AGP基因和B）AT基因表达的组织特异性。 实验方法如下：1）结构 将分离高和低诱导型AGP和AT的基因。 2)的转录活性和调节反应， 不同的AGP基因将通过将这些基因导入 人和大鼠肝癌细胞对这两种药物有反应， 肝脏急性期反应的最有效介质， 肝细胞刺激因子和糖皮质激素。 以识别 AT基因表达的肝肾特异性， 活动的M. 将检测并比较caroli AT基因 到M。 家犬AT基因转染肝脏和肝脏 肾组织培养细胞 在演示了正确的调节后 在AGP和AT基因中，顺式作用调节元件将 相对于结构基因而言是局部的。 3)在多大程度 哪些功能性AGP和AT基因序列支持正确的 组织、激素和发育调节将在 转基因小鼠 4)最后，细胞因子相互作用 特别是与顺式作用调节元件的结合将是 鉴定 这些研究结果表明，M. Caroli Genes将 为控制组织特异性的信号提供了新的见解， 和血浆蛋白基因的炎症诱导。