Multi-scale imaging of breast cancer proteins during DNA repair

项目摘要/摘要 乳腺癌易感蛋白BRCA1的突变与三联症的发生密切相关 阴性的乳腺癌和卵巢癌表型。目前，还没有精确的治疗方法来缓解 BRCA1基因突变对三阴性癌症患者的不良影响及其复发率 比任何其他形式的乳腺癌都要高。在细胞核中，BRCA1通过其自身的功能帮助保护基因组 参与DNA修复过程，从而起到肿瘤抑制作用。然而，携带BRCA1的细胞 突变失去了正确修复DNA损伤和转录其基因组的能力。……的高潮 这些事件导致基因组不稳定，并最终诱发癌症。准确地说， 突变的BRCA1未能执行其功能仍不清楚。理解这些的分子基础 缺陷可能为开发BRCA1相关癌症的新靶向疗法提供重要的洞察力。 基于我们从生化实验、低温电子显微镜(EM)成像、 和分子建模例行公事，我们认为癌症发展的差异与形态有关 BRCA1蛋白结构的变化。简单地说，改变其结构元素的BRCA1突变 会降低它在DNA修复过程中与其他蛋白质结合的能力。为了在亚细胞研究这一想法 我们将利用从共焦显微镜到高分辨率冷冻的多尺度成像技术。 嗯。将这种可视化信息与严格的生化分析相结合，我们将开发出一种独特的 基于分子的方法描述BRCA1在癌症形成中的多方面作用。我们也期待着 定义新的机会，以分子收集的机械洞察为基础设计有针对性的疗法 科学探究的水平。

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.