Multi-scale imaging of breast cancer proteins during DNA repair

项目概要/摘要 乳腺癌易感蛋白BRCA 1的突变与三重性乳腺癌的发生密切相关。 阴性乳腺癌和卵巢癌表型。目前，还没有精确的治疗方法来减轻 BRCA 1突变在三阴性癌症患者中的有害影响，以及这种疾病的复发率 比其他任何形式的乳腺癌都要高。在细胞核中，BRCA 1通过其自身的功能帮助保护基因组。 参与DNA修复过程，从而作为肿瘤抑制因子。然而，携带BRCA 1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性并最终导致癌症诱导。的精确方式 突变的BRCA 1无法执行其功能仍不清楚。了解这些的分子基础 这些缺陷可以为开发BRCA 1相关癌症的新靶向疗法提供重要的见解。 根据我们从生化实验、冷冻电子显微镜（EM）成像、 和分子建模程序，我们相信癌症发展的差异与形态学有关。 BRCA 1蛋白质结构的变化。简单地说，BRCA 1中改变其结构元件的突变 会降低其在DNA修复过程中与其他蛋白质结合的能力。为了在亚细胞水平上研究这个想法， 水平，我们将利用多尺度成像技术，从共聚焦显微镜到高分辨率冷冻， EM.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 以分子为基础的方法来描绘BRCA 1在癌症形成中的多方面作用。我们也期望 根据在分子生物学会议上收集的机制见解，确定设计靶向治疗的新机会 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.