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.

阿霉素是治疗乳腺癌最有效和最广泛使用的化疗药物之一。然而，对这种蒽环类药物的耐药性很常见，导致治疗失败和预后不良。三阴性乳腺癌 (TNBC) 是一种多见于非裔美国人和拉丁裔女性的临床亚型 (1, 2)，其特点是蒽环类药物治疗后复发率较高且总生存期较低 (1, 3)。开发针对 TNBC 的靶向疗法，特别是那些对阿霉素无反应的疗法，是临床治疗这种侵袭性疾病的最紧迫任务 (4)。多种机制与阿霉素耐药有关，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) 以及肿瘤抑制因子 ATM 或 p53 的突变 (7-9)，但这些发现尚未得到证实。广泛翻译用于临床益处。最近对 1,000 多种乳腺癌进行全基因组测序的研究表明，肿瘤抑制基因 p53 的突变率为 43-62% (10, 11)，使其成为该癌症类型中最常见的突变基因之一。 p53 是 DNA 损伤诱导细胞凋亡所必需的，因此旨在提高 p53 突变乳腺癌细胞对基因毒性治疗的敏感性的疗法将非常有益。我们最近报道，DNA 修复蛋白 DNAPK 调节 p53 独立的细胞凋亡 (12)，指出了一种使 p53 突变肿瘤敏感的新途径。为了鉴定调节 p53 独立细胞死亡的其他基因，我们对 p53 突变癌细胞进行了全激酶组 siRNA 阿霉素敏化剂筛选。阿霉素存活基因在 G2/M 细胞周期调节、DNA 修复和细胞凋亡中发挥作用。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 独立的细胞凋亡或有丝分裂细胞死亡，并使乳腺肿瘤对阿霉素敏感。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 出色的研究环境结合起来。我们在该试点提案中的目标是： 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消耗是否会加剧阿霉素处理的 p53 缺陷三阴性乳腺癌细胞中的 DNA 损伤并增加细胞凋亡。我们将确定沉默阿霉素存活基因是否会影响细胞周期进程和 DNA 复制，并评估进展为有丝分裂细胞死亡的机制。 2. 在乳腺癌临床前模型中验证候选治疗靶点。与目标 1 并行，我们将确定乳腺癌异种移植物中阿霉素存活基因的敲低是否会改善对阿霉素的反应。结果将是经过机械和临床前验证的多柔比星耐药 TNBC 靶点。了解控制阿霉素敏感性的途径将确定乳腺癌治疗的新候选药物靶点，并将指出新的生物标志物来对患者进行分层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，这些发现将阐明治疗这些无反应的侵袭性肿瘤的新策略。