PROJECT 1

项目1

• 批准号: 10474975
• 负责人: Reuben S Harris
• 金额: $ 27.51万
• 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/10474975
• 关键词: Accounting; Address; 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; 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; Financial compensation; Fluorescence; Future; Generations; Genetic; Genomic Segment; Genomics; Goals; Knowledge; Lesion; Malignant Neoplasms; Mediating; Membrane Proteins; Mismatch Repair; Molecular; Mutagenesis; Mutation; Neoplasm Metastasis; 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 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，占 每年诊断的浸润性乳腺癌中有超过 75% ER 阳性的总体突变情况。 乳腺癌是多因素造成的，但 DNA 脱氨酶 APOBEC3B (A3B) 占原发性疾病碱基取代突变的近 20%，而转移瘤中超过 50% 的 A3B 在正常情况下不表达。 乳腺上皮细胞中 A3B 过度表达，并且在大多数乳腺癌中过度表达。 与 ER 阳性乳腺癌的不良临床结果相关，包括复发、转移和 我们的计划正在测试 A3B 抑制作为佐剂的总体假设。 主要治疗方案将有助于预防突变驱动的不健康结果，例如耐药性 项目 1 将直接促进协作计划的努力，以通过以下方式检验这一假设。 3 个具体目标 在目标 1 中，我们建议开发用于量化 A3B 介导的编辑的报告系统。 活细胞，包括一种创新的、可运输的报告基因 在一个潜在的应用中，该系统将实现这一点。 在一组候选乳腺癌细胞系中快速检测候选小分子 A3B 抑制剂 在目标 2 中，我们将通过所有计划组成部分的协调活动来开发化合物。 这些研究将描述正常细胞和乳腺癌细胞中蛋白质水平 A3B 调节的机制。 重点关注蛋白质组学数据集综合表征的蛋白质-蛋白质相互作用。 预计直接相互作用也将揭示潜在的可成药表面，用于合作研究 在目标 3 中，我们研究了化学探针（项目 2）、计算建模（核心 C）和结构生物学（项目 3）。 将解决 A3B 催化的基因组尿嘧啶损伤如何处理成无错误和致突变结果的问题 这些研究有可能揭示 DNA 中的分子依赖性。 A3B 催化的 DNA 损伤水平升高的乳腺肿瘤细胞特有的修复。 因此，项目 1 是整个计划的一个组成部分，因为它将提供创新的检测方法 量化活体乳腺癌细胞中的 A3B 活性，获得对监管和潜在的分子见解 可药物化的蛋白质表面，并揭示可能构成新机会的遗传依赖性 诊断和治疗的发展。