PROJECT 1

项目1

• 批准号: 10916617
• 负责人: Reuben S Harris
• 金额: $ 8.05万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-09 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10916617
• 关键词: Accounting; Adjuvant; Aging; Amino Acids; Base Excision Repairs; Biochemical; Biological Assay; Biology; Breast; Breast Cancer Cell; Breast Cancer cell line; Breast Epithelial Cells; CDK4 gene; CRISPR screen; Cell Line; Cell Survival; Cells; Chemicals; Clinical; Collaborations; Compensation; Complex; Computer Models; Cytosine; DNA; DNA Damage; DNA Repair; DNA Repair Pathway; DNA glycosylase; Data Set; Deaminase; Deamination; Dependence; Development; Diagnosis; Diagnostic; Dinucleoside Phosphates; Disease; Drug resistance; Enzymes; Estrogen receptor positive; Event; Fluorescence; Future; Generations; Genetic; Genetic Transcription; Genomic Segment; Genomics; Goals; Knowledge; Lesion; Malignant Neoplasms; Mediating; Membrane Proteins; Methylation; Mismatch Repair; Molecular; Mutagenesis; Mutation; Neoplasm Metastasis; Nucleotides; Outcome; PIK3CA gene; Pathologic; Pathway Analysis; Pathway interactions; Physiological; Primary Neoplasm; Process; Proteins; Proteomics; RNA Splicing; Real-Time Systems; Recurrence; Regulation; Reporter; Reporting; Resistance; Role; Single-Stranded DNA; Source; Surface; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Uracil; antiviral immunity; base; base editing; breast cancer diagnosis; cancer cell; experimental study; homologous recombination; inhibitor; innovation; insight; malignant breast neoplasm; new technology; overexpression; preference; prevent; programs; protein protein interaction; rapid testing; small molecule; structural biology; technology development; therapeutic development; tumor

PROJECT 1 - BIOLOGY OF DNA DEAMINASES IN BREAST CANCER ABSTRACT Estrogen receptor (ER)-positive breast cancer is the most common form of breast cancer, accounting for over 75% of invasive breast cancers diagnosed each year. The overall mutation landscape in ER-positive breast cancer is multifactorial, but the DNA deaminase APOBEC3B (A3B) accounts for nearly 20% of base-substitution mutations in primary disease and over 50% in metastases. A3B is not expressed in normal mammary epithelial cells and becomes overexpressed in the majority of breast cancers. A3B overexpression correlates with poor clinical outcomes for ER-positive breast cancer, including recurrence, metastasis, and drug resistance. Our Program is testing the overarching hypothesis that A3B inhibition, as an adjuvant to primary treatment options, will help to prevent detrimental mutation-driven outcomes such as drug resistance and metastasis. Project 1 will contribute directly to collaborative Program efforts to test this hypothesis through 3 specific aims. In Aim 1, we propose to develop reporter systems for quantifying A3B-mediated editing in living cells, including an innovative, transportable reporter. In one potential application, this system will enable rapid testing of candidate small molecule A3B inhibitors in a panel of breast cancer cell lines as candidate compounds are developed through the concerted activities of all Program components. In Aim 2, we will delineate mechanisms of protein-level A3B regulation in normal and breast cancer cells. These studies will focus on protein-protein interactions prioritized by proteomics data sets. Comprehensive characterization of direct interactions is also anticipated to reveal potentially druggable surfaces for collaborative studies on chemical probes (Project 2), computational modeling (Core C), and structural biology (Project 3). In Aim 3, we will address how A3B-catalyzed genomic uracil lesions are processed into error-free and mutagenic outcomes by different DNA repair pathways. These studies have the potential to reveal molecular dependencies in DNA repair that are specific to breast tumor cells undergoing elevated levels of DNA damage catalyzed by A3B. Thus, Project 1 is an integral component of this overall Program because it will provide innovative assays for quantifying A3B activity in living breast cancer cells, yield molecular insights into regulatory and potentially druggable protein surfaces, and uncover genetic dependencies that may constitute new opportunities for diagnostic and therapeutic development.

项目1-乳腺癌中DNA脱氨酶的生物学 抽象的 雌激素受体（ER） - 阳性乳腺癌是最常见的乳腺癌形式 每年有超过75％的侵入性乳腺癌被诊断出。 ER阳性的总体突变景观 乳腺癌是多因素的，但是DNA脱氨酶Apobec3b（A3B）占基础替代的近20％ 原发性疾病的突变和转移酶的50％以上。 A3B未在正常情况下表达 乳腺上皮细胞并在大多数乳腺癌中过表达。 A3B过表达 与ER阳性乳腺癌的临床结局不良相关，包括复发，转移和 耐药性。我们的计划正在检验总体假设，即A3b抑制作用，作为辅助者 主要的治疗选择将有助于防止有害突变驱动的结果，例如耐药性 和转移。项目1将直接为合作计划努力做出贡献，以通过 3个具体目标。在AIM 1中，我们建议开发用于量化A3B介导的编辑的记者系统 活细胞，包括创新的可运输记者。在一个潜在的应用中，该系统将启用 快速测试候选小分子A3B抑制剂在一系列乳腺癌细胞系中作为候选者 化合物是通过所有程序组件的协同活动开发的。在AIM 2中，我们将 描述正常和乳腺癌细胞中蛋白质水平A3B调节的机制。这些研究会 专注于蛋白质组学数据集优先级的蛋白质 - 蛋白质相互作用。全面表征 预计直接相互作用也将揭示有关协作研究的潜在可吸毒表面 化学探针（项目2），计算建模（核心C）和结构生物学（项目3）。在AIM 3中，我们 将解决如何将A3B催化的基因组尿嘧啶病变处理为无误和诱变结果 通过不同的DNA修复途径。这些研究有可能揭示DNA中的分子依赖性 由A3B催化的DNA损伤水平升高的乳腺肿瘤细胞的修复。 因此，项目1是该总体程序的组成部分，因为它将为 量化活着的乳腺癌细胞中的A3b活性，产生分子见解，并可能 可吸毒蛋白质表面以及发现可能构成新机会的遗传依赖性 诊断和治疗发展。