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：
8392106
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 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8392106
关键词：
Adriamycin PFS Adverse effects Antibodies Apoptosis Apoptotic Attenuated BRCA1 gene BRCA2 gene Binding Biological Assay Breast Breast Cancer Cell Breast Cancer Treatment Breast Carcinoma Cancer Cell Growth Caspase Cell Cycle Cell Death Cell Line Cell Survival Cells Development Disease Dose Drug Kinetics Drug resistance Effectiveness Epidermal Growth Factor Receptor Epitopes Estrogen Receptors Etoposide Family member Fluorescence Polarization Gefitinib Goals Growth Healthcare Human In Vitro Intravenous Investigation Knowledge Laboratories Lead MAPK14 gene MCF7 cell Malignant Neoplasms Maps Mission Mitotic Molecular Molecular Target Mus New Agents Nuclear Oncogenic Pathway interactions Pharmaceutical Preparations Phenotype Progesterone Receptors Protein p53 Proteins Proteome Reagent Resistance Resistance development Retinoic Acid Receptor Risk Factors SCID Mice Signal Transduction Tail Tamoxifen Testing Therapeutic Therapeutic Agents Transducers Tumor Suppressor Proteins Ubiquitin Veins Veterans Woman Work Xenograft Model Xenograft procedure Yeasts acronyms analog anaphase-promoting complex base cancer cell cancer therapy caspase-9 cell growth cellular transduction chemotherapy combat cyclin B1 design drug development expectation fighting genetic regulatory protein high throughput screening human MAPK14 protein in vivo inhibitor/antagonist innovation insight loss of function malignant breast neoplasm mimetics multicatalytic endopeptidase complex mutant novel outcome forecast prevent protein function response small molecule small molecule libraries stress activated protein kinase therapeutic target treatment strategy tumor ubiquitin-protein ligase upstream kinase yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): The molecular complexity of breast cancers and therapy-associated side effects often limit effectiveness of many therapies, warranting the development of new agents for specific molecular targets while minimizing the off-target effects. Our goal is to develop novel, safer, and effective human breast cancer (HBC) therapies by exploiting functions of an apoptosis regulatory protein CCAR1/CARP-1. CARP-1 functions to transduce cell growth as well as chemotherapy (adriamycin, etoposide, or Iressa)-dependent inhibitory signaling. CARP-1 expression correlates inversely with HBC tumor grades. A yeast-two-hybrid (Y2H) screen revealed that CARP- 1 binds with the E3 ubiquitin ligase subunit anaphase promoting complex (APC)-2 protein. Following mapping of the interacting epitopes of CARP-1 and APC-2, we developed a fluorescence polarization assay (FPA) and screened a chemical library to identify small molecule inhibitors (SMIs) of CARP-1:APC-2 binding. The lead SMI termed CARP-1 Functional Mimetic (CFM)-4, inhibits HBC growth in vitro and in vivo. CFM-4 also attenuates growth of tamoxifen (TAM) or adriamycin (ADR)-resistant HBC cells but does not suppress growth of the non-tumorigenic and immortalized MCF-10A cells. CFM-4 binds with CARP-1, leads to increased CARP- 1 levels, activates pro-apoptotic p38 mitogen-activated protein kinase (MAPK), caspases-9 and 3, and apoptosis. CFM-4 induces loss of mitotic cyclin B1 and Cdc20 proteins, possibly through caspase-dependent but ubiquitin proteasome pathway (UPP)-independent mechanism(s). Hypothesis: CARP-1, a peri-nuclear phospho-protein, functions to regulate HBC cell growth and apoptosis, and is a key regulator of ADR signaling. We further hypothesize that CFM-4 stimulation of CARP-1 levels will provide a novel mechanism for treating breast and other cancers. Objectives: Our long-term objective is to elucidate mechanisms of CARP-1-dependent HBC growth inhibition, and to exploit this knowledge to develop agents for targeting of HBC. The rationale for this objective is based on the facts that loss of tumor suppressor p53 promotes development of aggressive HBCs and that resistance to ADR continues to be a problem. Since CARP-1 regulates HBC growth inhibition by ADR and is a co- activator of p53, the CFMs that bind and elevate CARP-1, activate pro-apoptotic p38 MAPK and caspases, and suppress Cdc20 and cyclin B1 levels have potential to target HBCs including those that have mutant p53 as well as have resistance to ADR or TAM. We will test our hypothesis by pursuing the following objectives. (1) To investigate molecular mechanism(s) of p38 activation, and the extent to which CARP-1 binding with p38 regulates HBC growth in the presence of CFM-4 or ADR. (2) To elucidate mechanism(s) of CARP-1-dependent activation of caspases-9 and -3, and determine the extent to which activated caspases target Cyclin B1 in the presence of CFM-4. (3) To conduct pharmacokinetic (PK) profiling and to demonstrate the anti-tumor activities of CFM(s) in mouse xenograft models of HBC. Potential Impact on Veterans Health Care: This investigation will facilitate development of strategies to prevent/combat breast cancer that will benefit women in the VA workforce, contribute to Veterans Health Care and further the mission of the VA.

描述（由申请人提供）：乳腺癌的分子复杂性和与治疗相关的副作用通常会限制许多疗法的有效性，从而为特定分子靶标的新药物开发，同时最大程度地减少脱靶效应。我们的目标是通过利用凋亡调节蛋白CCAR1/CARP-1的功能来开发新颖，更安全和有效的人类乳腺癌（HBC）疗法。鲤鱼1功能可转导细胞生长以及化学疗法（阿霉素，依托泊苷或Iressa）依赖性抑制性信号传导。 Carp-1表达与HBC肿瘤等级相关。酵母2杂交（Y2H）筛选显示，鲤鱼1与E3泛素连接酶亚基促进复合物（APC）-2蛋白结合。在映射Carp-1和APC-2的相互作用表位之后，我们开发了荧光极化测定法（FPA），并筛选了化学文库，以鉴定Carpc-1：APC-2结合的小分子抑制剂（SMIS）。铅SMI称为Carp-1功能模拟物（CFM）-4，在体外和体内抑制HBC的生长。 CFM-4还减轻了他莫昔芬（TAM）或adrimycin（ADR）耐药的HBC细胞的生长，但不能抑制非肿瘤和永生的MCF-10A细胞的生长。 CFM-4与鲤鱼1结合，导致鲤鱼1水平升高，激活促凋亡P38丝裂原激活的蛋白激酶（MAPK），CASPASE-9和3和凋亡。 CFM-4诱导有丝分裂蛋白B1和CDC20蛋白的丧失，可能是通过caspase依赖性但泛素蛋白酶体途径（UPP）独立的机理（S）的丧失。假设：鲤鱼1，一种核糖周磷酸蛋白，可调节HBC细胞生长和凋亡，是ADR信号传导的关键调节剂。我们进一步假设CFM-4刺激CARP-1水平将为治疗乳腺癌和其他癌症提供新的机制。目标：我们的长期目标是阐明依赖鲤鱼1的HBC生长抑制的机制，并利用这些知识来开发靶向HBC的剂。该目标的基本原理是基于以下事实：抑制肿瘤p53的丧失促进了侵略性HBC的发展，而对ADR的抵抗仍然是一个问题。由于Carp-1调节ADR的HBC生长抑制，并且是p53的共同激活剂，因此粘合和升高Carp-1，激活促凋亡P38 MAPK和CASPASE的CFM，并抑制Cdc20和Cyclin b1水平具有靶向HBC的潜力，包括具有突变p53以及具有突变p53和ADR或TAM的HBC。我们将通过追求以下目标来检验我们的假设。（1）研究p38激活的分子机制，以及在CFM-4或ADR存在下与p38结合的鲤鱼1与p38结合的程度。（2）阐明capase-9和-3的鲤鱼1依赖性激活的机理，并确定在CFM-4存在下激活的caspase靶向细胞周期蛋白B1的程度。（3）进行药代动力学（PK）分析并证明HBC小鼠异种移植模型中CFM的抗肿瘤活性。对退伍军人卫生保健的潜在影响：这项调查将有助于制定预防/对抗乳腺癌的策略，从而使弗吉尼亚州劳动力中的妇女受益，为退伍军人的卫生保健做出贡献，并进一步促进弗吉尼亚州的任务。