CARP-1: A Potential Therapeutic Agent for Breast Cancer

CARP-1：乳腺癌的潜在治疗剂

• 批准号: 8922216
• 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
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8922216
• 关键词: Adriamycin PFS; Adverse effects; Antibodies; Apoptosis; Apoptosis Regulation Gene; Apoptotic; Attenuated; Binding; Biology; Breast; 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; Epithelial; Etoposide; Female; Formulation; Gefitinib; Goals; Growth; Healthcare; Human; In Vitro; Knowledge; Laboratories; Lead; Lipids; MAPK8 gene; MAPK9 gene; MCF10A cells; Malignant Neoplasms; Mammary gland; Mediating; Mission; Molecular; N-terminal; Nuclear; Outcome; Pathway interactions; Phosphotransferases; Precipitation; Proteins; Proteome; Reagent; Resistance; 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; migration; mimetics; mutant; nano; novel; prevent; public health relevance; scaffold; small molecule inhibitor; small molecule libraries; targeted treatment; triple-negative invasive breast carcinoma; tumor; ubiquitin-protein ligase; yeast two hybrid system

 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)： 尽管最近在临床治疗方面取得了进展，但三阴性乳腺癌(TNBC)的分子复杂性和与治疗相关的副作用往往限制了许多治疗的有效性。制定新的和改进的TNBC治疗战略仍然是当务之急。我们先前发现了一个凋亡调节蛋白CARP-1/CCAR1。CARP-1的功能是转导细胞生长和化疗(阿霉素、依托泊苷或易瑞沙)依赖的抑制信号。CARP-1的表达与乳腺癌肿瘤分级呈负相关。酵母双杂交筛选和免疫共沉淀分析表明，CARP-1与后期促进复合体(APC/C)E3泛素连接酶亚基APC-2、细胞骨架丝蛋白C和细胞凋亡转导蛋白DEDD2结合。高通量筛选的化学文库产生了CARP-1：APC-2结合的抑制剂，称为CARP-1功能模拟(CFMS)。先导化合物CFM-4与CARP-1结合，刺激CARP-1表达和细胞凋亡。CFM-4在体内外均能抑制TNBC细胞的生长。CFM-4还抑制对他莫昔芬()或阿霉素(ADR)耐药的乳腺癌细胞的生长，而不抑制非致瘤和永生化的乳腺上皮MCF-10A细胞的生长。我们正在进行的研究进一步揭示：(1)CARP-1是丝氨酸/c-Jun氨基末端激酶(JNK)蛋白质组的一部分，JNK2调节ADR或CFM-4诱导的细胞凋亡，(2)CFM-4类似物CFM-4.16增强对TNBC细胞的ADR抑制，(3)CFM-4及其类似物部分通过上调CARP-1和DEDD2的水平，通过激活JNKs和caspase-8/9/3促进细胞凋亡，从而减弱TNBC细胞的迁移和侵袭，以及(4)CALP-1耗竭阻断CFMS或ADR对乳腺癌细胞的生长抑制。假说：CARP-1是一种核周磷酸蛋白，是ADR或CFM诱导细胞凋亡的关键调节因子，这些药物对CARP-1及其信号的调控是治疗TNBCs和其他乳腺癌的新机制。目的：阐明CARP-1依赖的乳腺癌生长抑制机制，并利用这一知识开发抗乳腺癌药物是我们的长期目标。P53基因缺失促进了侵袭性乳腺癌的发生，耐药肿瘤细胞的发生在临床上仍是一个棘手的问题。CARP-1是P53的共激活剂，而结合和上调CARP-1的CFM也抑制耐药的乳腺癌细胞，CARP-1的促凋亡功能可以弥补P53的丢失，特别是在缺乏功能性P53的TNBCs及其耐药变异体中。结合我们上面列出的简短发现，目前的初步研究通过以下几个方面为验证我们的假设提供了强有力的理论基础：(1)研究丝胺-鲤鱼-1结合的分子机制(S)，并确定依赖JNK/CARP-1的细胞凋亡调节TNBC生长的程度，以实现ADR或CFM的最佳疗效。(2)阐明CARP-1与DEDD2相互作用的机制(S)，并确定这种相互作用在多大程度上调节caspase-8/10的激活，并促进CFM或ADR诱导的细胞凋亡。(3)单独或与ADR合用CFM类似物纳米脂制剂后，确定CARP-1依赖的JNK/Caspase-8信号通路是否调节体内TNBC的生长。对退伍军人医疗保健的潜在影响：拟议的研究将产生宝贵的知识，以促进制定抗击乳腺癌的新的有效战略，使退伍军人管理局的女性劳动力受益，为退伍军人医疗保健做出贡献，并促进退伍军人管理局的使命。