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）： 尽管最近在临床管理方面取得了进展，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治疗的有效性。开发新的和改进的TNBC治疗策略仍然是紧迫的。我们以前发现了一个凋亡调节蛋白CARP-1/CCAR 1。CARP-1的功能是抑制细胞生长以及化疗（阿霉素、依托泊苷或易瑞沙）依赖性抑制信号传导。CARP-1表达与乳腺癌肿瘤分级呈负相关。酵母双杂交筛选和共免疫沉淀分析显示，CARP-1与后期促进复合物（APC/C）E3泛素连接酶亚基APC-2、细胞骨架骨架细丝蛋白C和凋亡转导DEDD 2蛋白结合。化学文库的高通量筛选产生CARP-1：APC-2结合的抑制剂，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表达和凋亡。CFM-4在体外和体内抑制TNBC细胞生长。CFM-4还减弱他莫昔芬（TAM）或阿霉素（ADR）抗性乳腺癌细胞的生长，而不抑制非致瘤性和永生化乳腺上皮MCF-10A细胞的生长。我们正在进行的研究进一步揭示了（1）CARP-1是细丝蛋白/c-jun N-末端激酶（JNK）蛋白质组的一部分，JNK 2通过ADR或CFM-4调节细胞凋亡;（2）CFM-4.16，一种新的CFM-4类似物，增强ADR对TNBC细胞的抑制作用;（3）CFM-4及其类似物部分通过升高CARP- 1和DEDD 2水平起作用，并通过激活JNK和半胱天冬酶-8/9/3促进凋亡，并减弱TNBC细胞的迁移和侵袭，以及（4）CARP-1缺失阻断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，是ADR或CFM引起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。目的：阐明CARP-1依赖性乳腺癌生长抑制的机制，并利用这一知识开发抗乳腺癌药物是我们的长期目标。p53的缺失促进了侵袭性乳腺癌的发展，并且ADR耐药的TNBC的发展仍然是临床上的一个棘手问题。事实上，CARP-1是p53的共激活剂，而结合和升高CARP-1的CFM也抑制ADR抗性乳腺癌细胞，CARP-1的促凋亡功能可以补偿p53的损失，特别是在TNBC及其缺乏功能性p53的耐药变体中。与我们上面列出的简要发现一起，目前的初步研究提供了强有力的理由来通过以下方式检验我们的假设：（1）研究细丝蛋白-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，以获得ADR或CFM的最佳功效。(2)阐明CARP-1与DEDD 2相互作用的机制，并确定这种相互作用调节半胱天冬酶-8/10活化的程度，并通过CFM或ADR促进细胞凋亡。(3)确定在单独或与ADR组合施用CFM类似物的纳米脂质制剂后，CARP-1依赖性JNK/胱天蛋白酶-8信号传导是否调节体内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，以促进新的和有效的战略，以打击乳腺癌的发展，这将有利于女性退伍军人的劳动力，有助于退伍军人医疗保健和进一步的退伍军人的使命。