17 Beta-estradiol induced caspase inhibitory factor

17 β-雌二醇诱导的 caspase 抑制因子

• 批准号: 6623859
• 负责人: ANDREA C LEBLANC
• 金额: $ 12.83万
• 依托单位: MCGILL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-15 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6623859
• 关键词: Alzheimer's disease; MCF7 cell; affinity chromatography; apoptosis; astrocytes; clinical research; complementary DNA; cysteine endopeptidases; enzyme activity; enzyme inhibitors; estradiol; gene expression; genetic regulation; hormone regulation /control mechanism; human tissue; immunocytochemistry; ion exchange chromatography; microinjections; molecular cloning; neurons; neuroprotectants; polymerase chain reaction; protein sequence; recombinant proteins

DESCRIPTION (provided by applicant): The primary goal of this application is to isolate a caspase inhibitory factor induced by 17-beta-estradiol in primary cultures of human neurons. A group of mammalian cysteinyl caspases is activated in a cell-, insult- and species-specific manner during apoptosis of various cell types. In human neurons, caspase-6 is active during serum deprivation-mediated neuronal apoptosis. We have previously shown that caspase-6 activity is lethal to human neurons in culture. Now, we find that 17 -beta-estradiol but not 17-alpha-estradiol, testosterone, or epitestosterone delay caspase-6 mediated neuronal cell death (Zhang et al. 2001). 17-beta-estradiol-treated neuronal extracts directly inhibit recombinant active caspase-6 in an in vitro assay. In contrast, 17-beta-estradiol does not induce CIF nor prevent caspase-mediated cell death in astrocytes. We conclude that 17-beta-estradiol induces a caspase inhibitory factor (CIF) that is preventing neuronal apoptosis. CIF is induced through estrogen receptors via a non-genomic pathway. We show that CIP is a broad spectrum caspase inhibitor between 10 and 14 kDa in size that is fairly resistant to boiling and proteinase K in neuronal extracts. Our results indicate that 17-beta-estradiol induces a novel inhibitor of active caspases and provide an additional mechanism for the neuroprotective action of 17-beta-estradiol. In this proposal, the primary goal is to identify CIF and determine its role in neuronal survival and cell death. In aim #1, we will biochemically isolate and sequence CIF. In aim #2, we will clone CIF cDNA and obtain antibodies. We will then confirm the role of CIF in neuronal survival and against caspases (aim #3), and determine the mode of activation of CIF (aim #4) and inactivation of caspase-6 (aim #5). Finally, we will study the regulation of CIF expression in normal and AD brains (aim #6). Given the strong epidemiological link of estrogen against Alzheimer's disease and its possible prophylactic role in neuroprotection, our results suggest a novel mechanism of action of 17-Beta-estradiol that could be exploited to promote neuroprotection in injury or neurodegenerative diseases.

描述（申请人提供）：此申请的主要目标是 孤立的17β-雌二醇诱导的胱天冬酶抑制因子 人类神经元的培养物。一组哺乳动物的半胱氨酸胱天蛋白酶被激活 在各种细胞凋亡过程中以细胞，侮辱和物种特异性的方式 细胞类型。在人类神经元中，caspase-6在血清期间活跃 剥夺介导的神经元凋亡。我们以前已经表明 caspase-6活性对培养中人类神经元具有致命性。现在，我们发现 那17-β-雌二醇，但不是17-α-雌二醇， 睾丸激素或表静脉蛋白延迟caspase-6介导的神经元细胞死亡 （Zhang等，2001）。 17β-雌二醇治疗的神经元提取物直接抑制 体外测定中的重组活性caspase-6。相反，17β-雌二醇 不会诱导CIF或防止星形胶质细胞中caspase介导的细胞死亡。我们 得出结论，17-β-雌二醇诱导caspase抑制因子（CIF） 预防神经元凋亡。 CIF通过雌激素受体诱导 非基因组途径。我们表明CIP是宽光谱caspase抑制剂 在10到14 kDa之间，对沸腾和 神经元提取物中的蛋白酶K。我们的结果表明17-β-雌二醇 诱导活性胱天蛋白酶的新型抑制剂，并提供额外 17β-雌二醇神经保护作用的机制。在此提案中， 主要目标是识别CIF并确定其在神经元存活中的作用 和细胞死亡。在AIM＃1中，我们将在生化分离株和序列CIF中。在 AIM＃2，我们将克隆CIF cDNA并获得抗体。然后，我们将确认 CIF在神经元生存和caspase中的作用（AIM＃3），并确定 CIF的激活方式（AIM＃4）和caspase-6的灭活方式（AIM＃5）。 最后，我们将研究正常和AD大脑中CIF表达的调节 （AIM＃6）。鉴于雌激素与阿尔茨海默氏症的牢固流行病学联系 疾病及其在神经保护中可能的预防作用，我们的结果 提出一种可以利用的17β-雌二醇的新型作用机理 促进损伤或神经退行性疾病中的神经保护作用。