Development Project 2

开发项目2

基本信息

批准号：
8744896
负责人：
BETI THOMPSON
金额：
$ 10.81万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-25 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8744896
关键词：
Affect African American Aliquot Anthracyclines Antibodies Apoptosis Apoptotic Attention Biochemical Bioinformatics Biological Assay Biological Markers Breast Breast Cancer Cell Bromodeoxyuridine Bypass Cancer Biology Cancer Model Cancer Patient Cancer cell line Candidate Disease Gene Cardiotoxicity Cell Cycle Cell Cycle Arrest Cell Cycle Checkpoint Cell Cycle Deregulation Cell Cycle Progression Cell Cycle Regulation Cell Death Cell Line Cell Proliferation Cells Chromosomal Instability Cleaved cell Clinical Clinical Treatment Consult Coupled Cyclin A Cyclin B DNA DNA Damage DNA Repair DNA biosynthesis DNA repair protein DNA-PKcs DNA-dependent protein kinase Data Data Analyses Detection Development Disease Dose Double-Blind Method Doxorubicin Doxycycline Drug Efflux Drug Targeting Ensure Environment Epidermal Growth Factor Receptor Epithelial Cells Evaluation Event Fiber Fibroblasts Flow Cytometry Fluorescence Freezing Gene Expression Gene Silencing Gene Targeting Genes Genetic Grant Head and Neck Squamous Cell Carcinoma Histology Human Immunoblot Analysis Immunoblotting Immunodeficient Mouse Immunohistochemistry In Vitro Individual Kinetochores Knowledge Laboratories Latina Lead Length Lentivirus Vector Letters M Phase Arrest M cell Maintenance Malignant neoplasm of ovary Mammary Neoplasms Measures Microscopy Mitosis Mitotic Molecular Mus Mutate Mutation Necrosis Neuroblastoma Normal Cell Outcome P-Glycoproteins PRKDC gene Pathway interactions Patients Phosphotransferases Pre-Clinical Model Preclinical Testing Promega Propidium Diiodide Protein p53 Proteins Protocols documentation Publishing RNA Randomized Reader Reagent Receptor Signaling Recurrence Reporting Research Research Personnel Resected Resistance Resolution Role Running S Phase Scientist Signal Transduction Small Interfering RNA Staging Staining method Stains Stress System Testing Tetanus Helper Peptide The Cancer Genome Atlas Therapeutic Time Topoisomerase II Topoisomerase Inhibitors Toxic effect Transfection Translating Travel Treatment Failure Tumor Suppressor Proteins Validation Woman Xenograft procedure anticancer research arm base cancer cell cancer therapy cancer type career development caspase-3 caspase-9 cell type chemotherapy clinical care cytotoxicity drinking water early onset experience genome sequencing graduate student improved in vivo insight interest luminescence malignant breast neoplasm matrigel micronucleus mortality mutant novel outcome forecast overexpression penis foreskin prevent promoter protein expression research study response screening small hairpin RNA therapeutic target therapy design triple-negative invasive breast carcinoma tumor tumor growth

项目摘要

Doxorubicin is one of the most effective and widely used chemotherapy agents for breast cancer. However, resistance to this anthracycline agent is common, leading to treatment failure and poor prognosis. Triple negative breast cancer (TNBC), a clinical subtype type seen disproportionately in African American and Latina women (1, 2), is characterized by higher recurrence and lower overall survival following anthracycline treatment (1, 3). Developing targeted therapies for TNBC, especially those that do not respond to doxorubicin, is the most urgent priority for the clinical treatment of this aggressive disease (4). Several mechanisms have been implicated in doxorubicin resistance, including an increase in drug efflux pathways (5), epidermal growth factor receptor signaling (6), and mutations in the tumor suppressors ATM or p53 (7-9), but these findings have not been widely translated for clinical benefit. Recent studies from whole genome sequencing of over 1,000 breast cancers reveal that the tumor suppressor p53 is mutated in 43-62% (10, 11), making it one the most commonly mutated genes in this cancer type. p53 is required for DNA damage induced apoptosis, so therapies designed to increase the sensitivity of p53 mutant breast cancer cells to genotoxic therapy would be immensely beneficial. We recently reported that the DNA repair protein DNAPK regulates p53 independent apoptosis (12), pointing to a novel pathway to sensitize p53 mutant tumors. To identity additional genes that regulate p53 independent cell death, we performed a kinome wide siRNA doxorubicin sensitizer screen with p53 mutant cancer cells. The doxorubicin survival genes identified function in G2/M cell cycle regulation, DNA repair, and apoptosis. Based on these findings, we hypothesize that targeting these doxorubicin survival genes will lead to p53 independent apoptotic or mitotic cell death and will sensitize breast tumors to doxorubicin. Our partnership combines expertise in DNA repair, cancer biology, and functional genetics together with the outstanding research environments of the FHCRC and NMSU. Our objectives in this pilot proposal are: 1. Identify mechanisms leading to p53 independent cell death. We will determine if depletion of doxorubicin survival genes exacerbates DNA damage and increases apoptosis in doxorubicintreated p53 deficient triple negative breast cancer cells. We will determine if silencing doxorubicin survival genes affects cell cycle progression and DNA replication, and assess the mechanism of progression into mitotic cell death. 2. Validate candidate therapeutic targets in preclinical models of breast cancer. In parallel with Aim 1, we will determine if knockdown of doxorubicin survival genes in breast cancer xenografts improves response to doxorubicin. The outcome will be mechanistically and preclinically validated targets for doxorubicin resistant TNBC. Knowledge ofthe pathways that control sensitivity to doxorubicin will identity new candidate drug targets for breast cancer therapy, and will point to new biomarkers to stratify patients and inform clinical care. Because doxorubicin resistance leads to treatment failure and subsequent mortality, these findings will elucidate new strategies to treat these unresponsive aggressive tumors.

阿霉素是乳腺癌最有效，最广泛使用的化学疗法之一。但是，对这种蒽环类药物的抗性很常见，导致治疗失败和预后不良。三重阴性乳腺癌（TNBC）是一种临床亚型类型，在非裔美国人和拉丁裔女性中（1，2）中观察到的临床亚型类型（1，2）的特征是蒽环类药物治疗后较高的复发和较低的总生存率（1，3）。开发针对TNBC的靶向疗法，尤其是那些对阿霉素反应的疗法，是这种侵略性疾病的临床治疗的最紧迫性（4）。多种机制已与阿霉素耐药性有关，包括药物外排途径的增加（5），表皮生长因子受体信号传导（6）和肿瘤抑制器ATM或p53（7-9）的突变，但这些发现并未被广泛翻译而获得临床利益。全基因组测序的最新研究超过1,000个乳腺癌表明，抑制肿瘤p53的43-62％（10，11）被突变，使其成为这种癌症类型中最常见的突变基因。 p53是DNA损伤引起的凋亡所必需的，因此旨在提高p53突变乳腺癌细胞对遗传毒性疗法的敏感性的疗法将非常有益。我们最近报道说，DNA修复蛋白DNAPK调节p53独立凋亡（12），指出了一种使p53突变肿瘤敏化的新途径。对于调节p53独立细胞死亡的其他基因，我们用p53突变癌细胞进行了宽siRNA siRNA siRNA sirna doxorubicin敏化剂。阿霉素存活基因在G2/M细胞周期调节，DNA修复和凋亡中鉴定出功能。基于这些发现，我们假设靶向这些阿霉素存活基因将导致p53独立的凋亡或有丝分裂细胞死亡，并将乳腺肿瘤敏感到阿霉素。我们的合作伙伴关系将DNA修复，癌症生物学和功能遗传学方面的专业知识与FHCRC和NMSU的杰出研究环境结合在一起。我们在该试点建议中的目标是：1。确定导致p53独立细胞死亡的机制。我们将确定阿霉素存活基因的耗竭是否加剧了DNA损伤并增加阿霉素治疗的p53 p53缺乏三重阴性乳腺癌细胞的凋亡。我们将确定沉默的阿霉素存活基因是否会影响细胞周期的进展和DNA复制，并评估发展为有丝分裂细胞死亡的机制。 2。验证乳腺癌临床前模型中的候选治疗靶标。与AIM 1同时，我们将确定乳腺癌异种移植中阿霉素存活基因的敲低是否可以改善对阿霉素的反应。该结果将在机械和临床上验证的阿霉素抗性TNBC的靶标。了解控制对阿霉素敏感性的途径将确定乳腺癌治疗的新候选药物靶标，并将指出新的生物标志物来分层患者并为临床护理提供信息。由于阿霉素耐药性会导致治疗失败和随后的死亡率，因此这些发现将阐明治疗这些无反应性攻击性肿瘤的新策略。