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）：乳腺癌是美国最常见的癌症之一，每八名妇女中就有一名在一生中患上这种疾病。除了是最普遍的癌症形式之一，它也是最具挑战性的治疗之一，每年有超过40，000名妇女死于这种疾病。乳腺癌治疗通常使用细胞毒性药物，导致癌细胞损伤，激活一种称为细胞凋亡的程序性细胞死亡形式。细胞凋亡的激活导致半胱氨酸蛋白酶（称为半胱天冬酶）的激活，其切割蛋白质底物并导致细胞的解体。细胞凋亡的抑制是乳腺癌的主要标志之一，而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。半胱天冬酶2（C2）已被证明是一个关键的启动凋亡的乳腺癌化疗药物。在一个模型实验系统中，我们的实验室先前已经表明，C2的激活可以通过结合小信号分子14-3-3？的代谢来调节。释放14-3-3号？是由14-3-3规定的吗乙酰化，并且这种修饰受细胞的代谢状态控制。随着广泛的研究表明，癌细胞通常有异常的代谢途径，我们建议，乳腺癌细胞调节乙酰化的14-3-3？才能在基因毒性压力下存活为了阐明如何改变乙酰化的14-3-3？可能有助于乳腺癌细胞逃避细胞死亡的细胞毒性药物的能力，我们建议确定酶调节14-3-3？乙酰化，代谢控制的机制，和14-3-3？正常和乳腺肿瘤样品中的乙酰化。此外，我们的目标是探索如何调节乙酰化将改变乳腺癌细胞的化学反应，无论是在文化和小鼠模型。拟议的研究将利用不同的化疗敏感性，从而建立乳腺癌细胞系，我们将使用特定的乙酰化酶和脱乙酰酶抑制剂筛选，以确定控制14-3-3？乙酰化然后通过监测不同代谢条件下酶结合和活性的变化来评价代谢控制的机制。此外，我们将确定是否乙酰化状态的14-3-3？通过分析正常和原发性人类肿瘤样品/切片中乙酰化14-3-3在乳腺癌中的改变？免疫组织化学。最后，为了确定调节乙酰化是否改变乳腺癌细胞的化学反应性，将相关抑制剂与化疗剂组合，并测定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。我们希望加强14-3-3？乙酰化将增加乳腺癌细胞对化疗诱导的细胞凋亡的易感性，潜在地导致发现可以与标准化疗组合用于更有效的乳腺癌治疗的新疗法。