Metabolic Regulation of 14-3-3zeta Acetylation in Breast Cancer

乳腺癌中 14-3-3zeta 乙酰化的代谢调节

• 批准号: 8451624
• 负责人: Erika Lee Segear Johnson
• 金额: $ 3.14万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8451624
• 关键词: Acetylation; Acetylesterase; Address; Apical; Apoptosis; Apoptotic; Binding; Binding Proteins; Biological Assay; Breast Cancer Cell; Breast Cancer Treatment; Breast Carcinoma; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Cancer cell line; Carbon; Caspase; Cell Death; Cell Proliferation; Cell Survival; Cell model; Cells; Cellular Stress; Cessation of life; Cleaved cell; Cysteine Protease; Cytotoxic agent; DNA Damage; Data; Deacetylase; Disease; Disease remission; Doxorubicin; Enzymes; Equilibrium; Etoposide; Experimental Models; Goals; Homeostasis; Human; Immunohistochemistry; Inhibition of Apoptosis; Lead; Malignant Neoplasms; Mammary Neoplasms; Mediating; Metabolic; Metabolic Control; Metabolic Pathway; Metabolism; Mitochondria; Modeling; Modification; Monitor; Mus; Mutagens; NADP; Normal tissue morphology; Nutrient; Oxidative Stress; Pathway interactions; Pentosephosphate Pathway; Peptide Hydrolases; Phagocytosis; Phosphorylation; Physiological; Post-Translational Protein Processing; Predisposition; Protein Dephosphorylation; Protein phosphatase; Proteins; Proteolysis; Regulation; Research; Resistance; Role; Sampling; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Stimulus; System; Tissues; United States; Woman; calmodulin-dependent protein kinase II; cancer cell; cancer therapy; caspase-2; chemotherapeutic agent; chemotherapy; cytochrome c; cytotoxic; effective therapy; inhibitor/antagonist; interest; malignant breast neoplasm; metaplastic cell transformation; mouse model; response; stressor; tumor; tumor xenograft

DESCRIPTION (provided by applicant): Breast cancer is one of the most prevalent forms of cancer in the United States, with one in every eight women developing the disease over the course of a lifetime. Besides being one of the most prevalent forms of cancer, it is also one of the most challenging to treat, with more than 40,000 women dying from the disease each year. Breast cancer therapies often utilize cytotoxic agents that cause damage to cancer cells, activating a form of programmed cell death called apoptosis. Activation of apoptosis results in the activation of cysteine proteases, known as caspases, which cleave protein substrates and lead to a disassembly of the cell. Inhibition of apoptosis is one of the central hallmarks of cance and the ability of cancer cells to evade cell death has been a major obstacle in breast cancer treatment. Caspase 2 (C2) has been shown to be a critical initiator of apoptosis in response to breast cancer chemotherapeutics. In a model experimental system, our lab has shown previously that activation of C2 can be regulated by metabolism through binding of a small signaling molecule, 14-3-3?. The release of 14-3-3? from C2 is regulated by 14-3-3? acetylation and this modification is controlled by the metabolic status of the cell. With extensive studies showing that cancer cells commonly have abnormal metabolic pathways, we propose that breast cancer cells modulate acetylation of 14-3-3? to survive genotoxic stressors. To elucidate how alterations in acetylation of 14-3-3? may contribute to the ability of breast cancer cells to evade cell death by cytotoxic agents, we propose to determine the enzymes regulating 14-3-3? acetylation, the mechanism of metabolic control, and the status of 14-3-3? acetylation in normal and breast tumor samples. Furthermore, we aim to explore how modulating acetylation will alter the chemoresponsiveness of breast cancer cells, both in culture and mouse models. The proposed research will utilize established breast cancer cell lines of varying chemosensitivities, whereby we will screen using specific acetylase and deacetylase inhibitors to identify the enzymes controlling 14-3-3? acetylation. The mechanism of metabolic control will then be evaluated by monitoring changes in enzyme binding and activity under different metabolic conditions. In addition, we will determine if the acetylation status of 14-3-3? is altered in breas cancer by analyzing both normal and primary human tumor samples/sections for acetylated 14-3-3? using immunohistochemistry. Finally, to determine whether modulating acetylation alters the chemoresponsiveness of breast cancer cells, relevant inhibitors will be combined with chemotherapeutic agents and assayed for their ability to induce cell death in both breast cancer cell lines and xenografted tumors. We expect that enhancing 14-3-3? acetylation will increase the susceptibility of breast cancer cells to chemotherapy-induced apoptosis, potentially leading to the discovery of a new therapy that can be used in combination with standard chemotherapies for more effective breast cancer treatments.

描述（由申请人提供）：乳腺癌是美国最普遍的癌症形式之一，一生中八分之一的妇女每八名妇女患这种疾病。除了是最普遍的癌症形式之一外，它也是最具挑战性的治疗之一，每年有40,000多名妇女死于该疾病。乳腺癌疗法经常利用细胞毒性剂对癌细胞造成损害，激活一种称为凋亡的程序性细胞死亡形式。凋亡的激活导致半胱氨酸蛋白酶（称为胱天蛋白酶）的激活，该蛋白酶裂解蛋白质底物并导致细胞拆卸。抑制细胞凋亡是CANCE的核心标志之一，癌细胞逃避细胞死亡的能力一直是乳腺癌治疗的主要障碍。 caspase 2（C2）已被证明是响应乳腺癌化学治疗药的凋亡的关键起始人。在模型实验系统中，我们的实验室先前已经表明，C2的激活可以通过小型信号分子的结合来调节代谢，14-3-3？ 14-3-3的发布？从C2受到14-3-3的调节？乙酰化和这种修饰受细胞的代谢状态控制。通过广泛的研究表明，癌细胞通常具有异常的代谢途径，我们提出乳腺癌细胞调节14-3-3的乙酰化？生存的遗传毒性应激源。阐明乙酰化的改变是14-3-3的如何变化？我们可能有助于乳腺癌细胞通过细胞毒性剂逃避细胞死亡的能力，我们建议确定调节14-3-3的酶？乙酰化，代谢控制的机制和14-3-3的状态？正常和乳腺肿瘤样品中的乙酰化。此外，我们旨在探讨如何调节乙酰化将如何改变培养和小鼠模型中乳腺癌细胞的化学回应性。拟议的研究将利用变化的化学固定性的已建立的乳腺癌细胞系，我们将使用特定的乙酰基酶和脱乙酰基酶抑制剂进行筛查，以鉴定控制14-3-3的酶？乙酰化。然后，将通过监测不同代谢条件下酶结合和活性的变化来评估代谢控制的机制。此外，我们将确定14-3-3的乙酰化状态是否？通过分析乙酰化14-3-3的正常和原发性人类肿瘤样品/切片是否会改变bre虫癌的改变？使用免疫组织化学。最后，为了确定调节乙酰化是否会改变乳腺癌细胞的化学回应性，相关抑制剂将与化学治疗剂结合使用，并根据其在乳腺癌细胞系和异种移植肿瘤中诱导细胞死亡的能力而进行了测定。我们希望增强14-3-3？乙酰化将增加乳腺癌细胞对化学疗法诱导的细胞凋亡的敏感性，这可能导致发现一种新疗法，该疗法可与标准化学疗法结合使用，以进行更有效的乳腺癌治疗。