Multi-scale imaging of breast cancer proteins during DNA repair

DNA 修复过程中乳腺癌蛋白的多尺度成像

• 批准号: 10183192
• 负责人: Deborah F Kelly
• 金额: $ 40.77万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10183192
• 关键词: Address; Affect; Architecture; BARD1 gene; BRCA1 Mutation; BRCA1 Protein; BRCA1 gene; Binding; Biochemical; Biological Assay; Breast Cancer Cell; Cancer Patient; Cell Nucleus; Cell Survival; Cells; Clinical; Co-Immunoprecipitations; Confocal Microscopy; Cryoelectron Microscopy; DNA; DNA Damage; DNA Repair; Data; Data Collection; Defect; Detection; Development; Disease; Electrons; Elements; Ensure; Estrogens; Event; Genetic Materials; Genetic Transcription; Genome; Genomic Instability; Hot Spot; Human; Image; Imaging Device; Imaging technology; Incidence; Inherited; Link; Malignant Neoplasms; Malignant neoplasm of ovary; Maps; Molecular; Molecular Structure; Morphology; Motion; Mutate; Mutation; Nuclear; Outcome; Pathology; Patients; Phenotype; Predisposition; Prevention; Process; Proteins; Publishing; RNA Polymerase II; Reagent; Recurrence; Repair Complex; Research; Resolution; Role; Scientific Inquiry; Structure; Structure-Activity Relationship; Testing; Time; Titan; Tumor Suppressor Proteins; Visualization; Western Blotting; Woman; Work; base; biochemical tools; breast imaging; cancer cell; cancer risk; carcinogenesis; clinically relevant; design; detector; experimental study; genome integrity; insight; live cell imaging; malignant breast neoplasm; microscopic imaging; molecular modeling; nanometer; new therapeutic target; oxidative damage; protein structure; repaired; response; targeted treatment; therapeutic target; tool; triple-negative invasive breast carcinoma

PROJECT SUMMARY/ABSTRACT Mutations in the breast cancer susceptibility protein, BRCA1, are heavily linked to the development of triple negative breast and ovarian cancer phenotypes. Currently, there are no precise treatments to mitigate the detrimental effects of BRCA1 mutations in triple negative cancer patients, and recurrence rates for this disease are higher than in any other form of breast cancer. In the nucleus, BRCA1 helps protect the genome by its involvement in DNA repair processes, thus serving as a tumor suppressor. However, cells harboring BRCA1 mutations lose the ability to properly repair DNA damage and transcribe their genome. The culmination of these events leads to genomic instability and eventually cancer induction. The precise manner in which mutated BRCA1 fails to execute its functions remains unclear. Understanding the molecular basis for these defects could provide significant insight toward developing new targeted therapies for BRCA1-related cancers. Based on our preliminary data derived from biochemical experiments, cryo-Electron Microscopy (EM) imaging, and molecular modeling routines, we believe differences in cancer development are related to morphological changes in the BRCA1 protein structure. Simply stated, mutations in BRCA1 that alter its structural elements will reduce its ability to engage other proteins during DNA repair. To investigate this idea at the subcellular level, we will utilize multi-scale imaging technology ranging from confocal microscopy to high-resolution cryo- EM. Combining this visualization information with rigorous biochemical analysis, we will develop a unique molecular-based approach to delineate the multifaceted role of BRCA1 in cancer formation. We also expect to define new opportunities to design targeted therapies based on mechanistic insights gathered at the molecular level of scientific inquiry.

项目摘要/摘要 乳腺癌易感性蛋白BRCA1的突变与三重的发展密切相关 负乳腺癌和卵巢癌表型。目前，没有精确的治疗方法来减轻 BRCA1突变在三重阴性癌症患者中的有害作用以及该疾病的复发率 高于任何其他形式的乳腺癌。在细胞核中，BRCA1有助于保护基因组 参与DNA修复过程，因此用作肿瘤抑制器。但是，带有BRCA1的细胞 突变失去了正确修复DNA损伤并转录其基因组的能力。的高潮 这些事件导致基因组不稳定性并最终导致癌症诱导。精确的方式 突变的BRCA1无法执行其功能尚不清楚。了解这些分子基础 缺陷可以为开发与BRCA1相关的癌症开发新的靶向疗法提供重大见解。 根据我们从生化实验得出的初步数据，冷冻电子显微镜（EM）成像， 和分子建模惯例，我们认为癌症发展的差异与形态学有关 BRCA1蛋白结构的变化。简而言之，BRCA1中改变其结构元素的突变 将降低其在DNA修复过程中参与其他蛋白质的能力。在亚细胞上调查这个想法 级别，我们将利用从共聚焦显微镜到高分辨率冷冻的多尺度成像技术 Em。将这些可视化信息与严格的生化分析相结合，我们将开发独特的 基于分子的方法来描述BRCA1在癌症形成中的多方面作用。我们也期望 根据在分子上收集的机械见解来定义设计有针对性疗法的新机会 科学探究水平。