Identification of small molecule activators of procaspase-7 to caspase-7

procaspase-7 至 caspase-7 小分子激活剂的鉴定

• 批准号: 7688986
• 负责人: Diana C West-Szymanski
• 金额: $ 4.1万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-16 至 2011-01-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7688986
• 关键词: Apoptosis; Apoptotic; Caspase; Caspase Inhibitor; Cell membrane; Cells; Cessation of life; Characteristics; Chromatin; Cleaved cell; Colon; Cysteine Protease; Development; Enzyme Precursors; Family; Flow Cytometry; Genus Cola; Goals; HL60; Immunohistochemistry; In Vitro; Laboratories; Libraries; MCF7 cell; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Monitor; Mutation; PAC1 phosphatase; Pan Genus; Paraffin Embedding; Pathway interactions; Physical condensation; Prostate; Proteins; Research; Serine; Staging; Staining method; Stains; Stimulus; Techniques; Time; Tissues; Western Blotting; cancer cell; cancer therapy; caspase-2; caspase-3; caspase-6; caspase-7; in vivo; interest; malignant breast neoplasm; pro-caspase-3; procaspase-7; small molecule; trait; tumor growth

DESCRIPTION (provided by applicant): One hallmark of cancer is the ability to evade programmed cell death, or apoptosis; this allows uncontrolled proliferation and tumor growth. Our goal is to use a small molecule to activate to trigger apoptosis in cancer cells by targeting key proteins in the apoptotic pathway: executioner procaspases. We hope to develop a strategy for personalized cancer treatment in which cancers with elevated levels of procaspases will be treated with specialized small molecules. It is known that there are elevated levels of procaspase-7 in certain cancers, such as prostate cancer. We hypothesize that apoptosis can be selectively induced in cancer cells containing elevated levels of procaspase-7 via a direct small molecule activator of procaspase- 7 to caspase-7. This hypothesis is supported by recent research in our group in which we were able to induce apoptosis in cancer cells by small molecule activation of procaspase-3 to caspase-3. Specific Aim 1. Identification of small molecules which activate procaspase-7 to caspase-7. In this specific aim we intend to A) perform timecourse analyses to elucidate how procaspase-7 activates over time, B)identify a small molecule activator of procaspase-7 from a library screen, and C) validate procaspase-7-activating compounds found in the library screen. Specific Aim 2. Induction of apoptotic death using the identified small molecule activator of procaspase-7. During apoptosis, cells display a variety of characteristic traits. We will use various techniques to monitor the apoptotic hallmarks in HL60 and MCF-7 cells induced by the identified small molecule in Specific Aim 1. Techniques include Flow cytometry to detect phosphatidyl serine exposure on apoptotic cell membranes, Hoescht staining to detect chromatin condensation in apoptotic cells, Rescue from apoptotic death with a pan-caspase inhibitor Specific Aim 3. Identification of specific cancers in which executioner procaspase levels are elevated. A variety of paraffin-embedded primary cancer tissues are available to the Hergenrother laboratory. Using Western blotting and immunohistochemistry, we can determine executioner procaspase-3, -6, and -7 levels in prostate and breast cancer tissues.

描述（由申请人提供）：癌症的一个标志是能够逃避程序性细胞死亡或细胞凋亡；这使得增殖和肿瘤生长不受控制。我们的目标是通过靶向凋亡途径中的关键蛋白：刽子手半胱天冬酶，使用小分子来激活并触发癌细胞凋亡。我们希望制定一种个性化癌症治疗策略，其中前天冬氨酸蛋白酶水平升高的癌症将用专门的小分子进行治疗。众所周知，某些癌症（例如前列腺癌）中 procaspase-7 水平升高。我们假设，通过 procaspase-7 到 caspase-7 的直接小分子激活剂，可以在含有升高水平的 procaspase-7 的癌细胞中选择性诱导细胞凋亡。这一假设得到了我们小组最近的研究的支持，其中我们能够通过小分子激活 procaspase-3 到 caspase-3 来诱导癌细胞凋亡。 具体目标 1. 鉴定将 procaspase-7 激活为 caspase-7 的小分子。在这个具体目标中，我们打算 A) 进行时间过程分析，以阐明 procaspase-7 如何随时间激活，B) 从文库筛选中识别 procaspase-7 的小分子激活剂，以及 C) 验证在文库筛选中发现的 procaspase-7 激活化合物。 具体目标 2. 使用已鉴定的 procaspase-7 小分子激活剂诱导细胞凋亡。在细胞凋亡过程中，细胞表现出多种特征。我们将使用各种技术来监测特定目标 1 中鉴定的小分子诱导的 HL60 和 MCF-7 细胞的凋亡标志。技术包括流式细胞术检测凋亡细胞膜上磷脂酰丝氨酸的暴露、Hoescht 染色检测凋亡细胞中的染色质浓缩、用泛半胱天冬酶抑制剂挽救细胞凋亡 具体目标 3. 鉴定刽子手天冬氨酸蛋白酶原水平升高的特定癌症。 Hergenrother 实验室可以获得各种石蜡包埋的原发癌组织。使用蛋白质印迹和免疫组织化学，我们可以确定前列腺癌和乳腺癌组织中的刽子手蛋白酶原 3、-6 和 -7 水平。