CARP-1: A Potential Therapeutic Agent for Breast Cancer

CARP-1：乳腺癌的潜在治疗剂

基本信息

批准号：
10025565
负责人：
Arun Kumar Rishi
金额：
--
依托单位：
JOHN D DINGELL VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-01-01 至 2020-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10025565
关键词：
Adriamycin PFS Antibodies Apoptosis Apoptosis Regulation Gene Apoptotic Attenuated Binding Biology Breast Cancer Cell Breast Cancer Treatment Breast Cancer therapy CASP8 gene Cancer Cell Growth Caspase Cell Line Cell Survival Cells Chemicals Clinic Clinical Clinical Management Co-Immunoprecipitations Development Disease Resistance Drug resistance Effectiveness Epidermal Growth Factor Receptor Estrogen receptor positive Etoposide Female Formulation Gefitinib Goals Growth Healthcare Human In Vitro Inhibition of Cancer Cell Growth Knowledge Laboratories Lead Lipids MAPK8 gene MAPK9 gene MCF10A cells Malignant Neoplasms Mediating Mission Molecular N-terminal Nuclear Outcome Pathway interactions Phosphoproteins Phosphotransferases Precipitation Proteins Proteome Reagent Resistance Resistance development Scaffolding Protein Signal Transduction Subgroup TP53 gene Tamoxifen Testing Therapeutic Agents Transducers Variant Veterans Western Blotting Work Xenograft procedure Yeasts acronyms analog anaphase-promoting complex base cell growth cell motility cellular transduction chemotherapy combat expectation filamin genetic regulatory protein high throughput screening improved in vivo inhibitor/antagonist innovation insight knock-down malignant breast neoplasm mammary epithelium migration mimetics mutant nano novel prevent public health relevance scaffold side effect small molecule inhibitor small molecule libraries targeted treatment triple-negative invasive breast carcinoma tumor ubiquitin-protein ligase yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Despite recent advances in clinical management, the molecular complexity of triple-negative breast cancers (TNBC) and therapy-associated side effects often limit effectiveness of many therapies. Development of new and improved strategies for TNBC treatment remains urgent. We previously discovered an apoptosis regulatory protein CARP-1/CCAR1. CARP-1 functions to transduce cell growth as well as chemotherapy (adriamycin, etoposide, or Iressa)-dependent inhibitory signaling. CARP-1 expression correlates inversely with breast cancer tumor grades. A yeast-two-hybrid screen together with co-immuno-precipitation analyses revealed that CARP-1 binds with the anaphase-promoting complex (APC/C) E3 ubiquitin ligase subunit APC-2, cytoskeletal scaffold filamin C , and apoptosis-transducing DEDD2 proteins. High-throughput screening of a chemical library yielded inhibitors of CARP-1:APC-2 binding termed CARP-1 functional mimetics (CFMs). Lead compound CFM-4 binds CARP-1, stimulates CARP-1 expression and apoptosis. CFM-4 inhibits TNBC cell growth in vitro and in vivo. CFM-4 also attenuates growth of tamoxifen (TAM) or adriamycin (ADR)-resistant breast cancer cells without inhibiting growth of the non-tumorigenic and immortalized mammary epithelial MCF-10A cells. Our on-going studies further revealed that (1) CARP-1 is a part of filamin/c-jun N-terminal kinase (JNK) proteome, and JNK2 regulates apoptosis by ADR or CFM-4, (2) CFM-4.16, a novel CFM-4 analog, enhances ADR inhibition of TNBC cells, (3) CFM-4 and its analogs function in part by elevating CARP- 1 and DEDD2 levels, and promote apoptosis by activating JNKs and caspases-8/9/3, and attenuate TNBC cell migration and invasion, and (4) CARP-1 depletion blocks breast cancer cell growth inhibition by CFMs or ADR. Hypothesis: CARP-1, a peri-nuclear phospho-protein, is a key regulator of apoptosis by ADR or CFMs, and that modulation CARP-1 and its signaling by these agents is a novel mechanism for treating TNBCs and other breast cancers. Objectives: Elucidation of mechanisms of CARP-1-dependent breast cancer growth inhibition, and exploitation of this knowledge to develop anti-breast cancer agents are our long-term goals. Loss of p53 promotes development of aggressive breast cancers and development of ADR-resistant TNBCs remains a formidable problem in clinic. The facts that CARP-1 is a co-activator of p53 while CFMs that bind and elevate CARP-1 also inhibit ADR-resistant breast cancer cells, apoptosis-stimulating functions of CARP-1 could compensate for loss of p53, especially in TNBCs and their drug-resistant variants that lack functional p53. Together with our brief findings listed above, the current preliminary studies provide a strong rationale to test our hypothesis by: (1) Investigating molecular mechanism(s) of filamin-CARP-1 binding, and determining the extent to which JNK/CARP-1-dependent apoptosis regulates TNBC growth for optimal efficacy of ADR or CFMs. (2) Elucidating mechanism(s) CARP-1 interaction with DEDD2, and determining the extent to which this interaction regulates activation of caspases-8/10, and contributes to apoptosis by CFMs or ADR. (3) Determining whether CARP-1-dependent JNK/Caspase-8 signaling regulates TNBC growth in vivo following administration of nano-lipid formulations of CFM analogs alone or in combination with ADR. Potential Impact on Veterans Health Care: The proposed studies will yield valuable knowledge to facilitate development of novel and effective strategies to combat breast cancer that will benefit female VA workforce, contribute to Veterans Health Care and further the mission of the VA.

描述（由申请人提供）：尽管临床管理最近取得了进展，但三阴性乳腺癌 (TNBC) 的分子复杂性和治疗相关的副作用往往限制了许多疗法的有效性，我们之前发现了细胞凋亡调节。蛋白 CARP-1/CCAR1。CARP-1 具有转导细胞生长以及化疗（阿霉素、依托泊苷或易瑞莎）依赖性信号传导的作用，CARP-1 的表达与乳腺癌肿瘤 A 级呈负相关。酵母双杂交筛选和免疫共沉淀分析表明，CARP-1 与后期促进复合物 (APC/C) E3 泛素连接酶亚基 APC-2、细胞骨架支架细丝蛋白 C 和细胞凋亡转导 DEDD2 蛋白结合高通量筛选化学库产生 CARP-1:APC-2 结合抑制剂，称为 CARP-1 功能模拟物(CFM)。先导化合物 CFM-4 结合 CARP-1，刺激 CARP-1 表达和细胞凋亡。CFM-4 还可抑制他莫昔芬 (TAM) 或阿霉素 (ADR) 的生长。）耐药乳腺癌细胞，而不抑制非致瘤和永生化乳腺上皮 MCF-10A 细胞的生长，我们正在进行的研究进一步表明（1）。 CARP-1 是 filamin/c-jun N 末端激酶 (JNK) 蛋白质组的一部分，JNK2 通过 ADR 或 CFM-4 调节细胞凋亡，(2) CFM-4.16 是一种新型 CFM-4 类似物，可增强 ADR 抑制TNBC 细胞，(3) CFM-4 及其类似物部分通过提高 CARP-1 和 DEDD2 水平发挥作用，并通过激活 JNK 和 DEDD2 促进细胞凋亡caspase-8/9/3，并减弱 TNBC 细胞迁移和侵袭，(4) CARP-1 耗竭可阻止 CFM 或 ADR 对乳腺癌细胞生长的抑制。假设：CARP-1 是一种核周磷蛋白。 ADR 或 CFM 细胞凋亡的关键调节因子，通过这些药物调节 CARP-1 及其信号传导是治疗 TNBC 和其他乳腺癌的新机制。阐明 CARP-1 依赖性乳腺癌生长抑制机制，并利用这些知识开发抗乳腺癌药物是我们的长期目标。p53 的缺失会促进侵袭性乳腺癌的发展，而 ADR 抗性 TNBC 的发展仍然如此。事实上，CARP-1 是 p53 的共激活剂，而结合并升高 CARP-1 的 CFM 也能抑制 ADR 抗性乳腺癌细胞、细胞凋亡刺激功能。 CARP-1 可以补偿 p53 的缺失，特别是在 TNBC 及其缺乏功能性 p53 的耐药变体中，结合我们上面列出的简要发现，当前的初步研究通过以下方式为检验我们的假设提供了强有力的理由：(1) 调查。丝蛋白-CARP-1 结合的分子机制，并确定 JNK/CARP-1 依赖性细胞凋亡调节 TNBC 生长的程度，以实现 ADR 或 CFM 的最佳功效 (2) 阐明。机制 CARP-1 与 DEDD2 相互作用，并确定这种相互作用调节 caspase-8/10 激活的程度，并通过 CFM 或 ADR 促进细胞凋亡 (3) 确定 CARP-1 是否依赖 JNK/Caspase。单独使用 CFM 类似物纳米脂质制剂或与 ADR 联合使用后，-8 信号传导可调节体内 TNBC 生长。对退伍军人医疗保健的潜在影响：拟议的研究将产生有价值的知识。促进制定新颖有效的乳腺癌防治策略，这将使退伍军人管理局女性劳动力受益，为退伍军人医疗保健做出贡献，并进一步实现退伍军人管理局的使命。