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）：尽管最近的临床管理进展，但三阴性乳腺癌（TNBC）的分子复杂性和与治疗相关的副作用通常限制了许多疗法的有效性。开发新的和改进的TNBC治疗策略仍然是紧急的。我们以前发现了调节蛋白质Carp-1/ccar1的细胞凋亡。 Carp-1的功能可以翻译细胞生长以及化学疗法（阿霉素，依托泊苷或IRESSA）依赖性抑制信号传导。 Carp-1表达与乳腺癌肿瘤成绩相关。酵母 - 两种杂交筛网以及共免疫的沉积分析表明，鲤鱼1与后期促进复合物（APC/C）E3泛素连接酶亚基APC-2，细胞骨架骨骼支架C和凋亡转化 - 转化 - 刺激性蛋白质的蛋白质结合。化学文库的高通量筛选产生了鲤鱼1的抑制剂：APC-2结合称为Carp-1功能模仿（CFMS）的抑制剂。铅化合物CFM-4结合鲤鱼1，刺激鲤鱼1表达和凋亡。 CFM-4在体外和体内抑制TNBC细胞的生长。 CFM-4还减轻了他莫昔芬（TAM）或阿霉素（ADR）耐药性乳腺癌细胞的生长，而不会抑制非肿瘤和永生化的乳腺上皮MCF-10A细胞的生长。 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鲤鱼1和DEDD2水平，并通过激活JNK和CASPASE-8/9/3促进凋亡，并减弱TNBC细胞迁移和侵袭，以及（4）Carp-1耗竭阻止CFMS或ADR抑制乳腺癌细胞的生长。假设：Carp-1是一种核周期磷酸蛋白，是ADR或CFMS凋亡的关键调节剂，这些药物的调节Carp-1及其信号传导是治疗TNBC和其他乳腺癌的新机制。目的：阐明依赖鲤鱼-1依赖性乳腺癌生长抑制的机制，并剥削这种知识来开发抗胸腺癌剂，这是我们的长期目标。 p53的丧失促进了侵略性乳腺癌的发展，而抗ADR的TNBC的发展仍然是诊所中的一个巨大问题。 Carp-1是p53的共激活因子的事实，而结合和升高鲤鱼1的CFM也抑制了耐ADR的乳腺癌细胞，Carp-1的凋亡刺激功能可以弥补p53的损失，尤其是在TNBC及其缺乏功能性p53的药物耐药变体中。与上面列出的简短发现一起，当前的初步研究提供了一个有力的理由来检验我们的假设：（1）研究Filamin-CARP-1结合的分子机制（S），并确定JNK/Carp-1依赖性凋亡的程度在多大程度上调节了TNBC TNBC TNBC的最佳ADR ADR效率。（2）阐明机制与DEDD2的相互作用，并确定这种相互作用调节caspases-8/10的激活程度，并导致CFMS或ADR的凋亡。（3）确定carp-1依赖性JNK/caspase-8信号传导是否单独使用CFM类似物的纳米脂质公式后，调节体内TNBC的生长，还是与ADR结合使用。对退伍军人卫生保健的潜在影响：拟议的研究将产生有价值的知识，以促进制定新颖有效的策略，以打击乳腺癌，从而使女性VA劳动力受益，为退伍军人卫生保健做出贡献，并进一步促进VA的使命。