(PQC2)Nanoscale changes in 3D nuclear architecture during breast tumorigenesis

这个拟议的项目是为了解决C组提出的挑衅性问题2：如何才能 肿瘤的属性，如细胞的电、光或机械属性，可用于 提供更早或更可靠的癌症检测、诊断、预后或药物监测 反应或肿瘤复发？我们将研究光学特性及其相关 癌变过程中细胞核的纳米尺度构筑变化及其确定 准确地提供乳腺癌的早期和更准确的诊断和预后。我们 假设核结构的纳米级改变发生在致癌的早期 以及容易获得的核纳米尺度变化的光学标记的测量 架构可以作为一种经济高效且准确的工具，用于更早、更准确地诊断癌症 诊断和预后。我们团队开发了一套光学显微镜系统，可以 全面表征核结构中的3D纳米级变化 使用临床获得的常规福尔马林固定和石蜡包埋组织致癌。 我们的光学显微镜系统包括深度分辨空域低相干性 定量位相显微镜(深度分辨SL-QPM)和空间光谱编码 频率(SESF)。我们证明了深度分辨的SL-QPM在一个 单个细胞核内1 nm的灵敏度，而SESF提取结构长度-尺度 分布的精确度为10-20 nm。我们广泛的初步数据表明， 这些光学标记有望检测浸润性癌症的存在，即使是在 来自多种肿瘤类型的组织学正常细胞，并预测癌症进展风险。现在 我们建议将这两个光学显微镜系统与最先进的3D技术结合使用 超分辨率显微镜，以定义一组光学标记和底层纳米级 核结构的改变是肿瘤发生和发展的每个阶段的特征 找出那些检测到“癌前”变化的基因。然后我们将进行临床研究，以 评估用于预测核结构中纳米尺度变化的光学标记的准确性 患有癌前病变(例如，非典型病变)的妇女的乳腺癌进展风险 导管原位癌(DCIS))和浸润性前癌(DCIS)以避免过度 治疗。如果这个项目成功，将建立纳米级核的改变 架构在癌症发生中的作用，并对肿瘤生物学研究和 临床护理。这将为未来纳米级光学标记的应用奠定坚实的基础 核结构的变化可作为准确的预后标志来预测那些 有可能进展为浸润性癌症。

This proposed project is to address provocative question 2 from Group C: "How can the physical properties of tumors, such as the cell's electrical, optical or mechanical properties, be used to provide earlier or more reliable cancer detection, diagnosis, prognosis, or monitoring of drug response or tumor recurrence?" We will investigate the optical properties and their associated nanoscale architectural changes in the cell nucleus during carcinogenesis and determine their accuracy in providing earlier and more accurate diagnosis and prognosis of breast cancer. We hypothesize that the nanoscale alterations in nuclear architecture occur early in carcinogenesis and the measurement of easily obtained optical markers of nanoscale changes in nuclear architecture can serve as a cost-effective and accurate tool for earlier and more accurate cancer diagnosis and prognosis. Our group has developed a set of optical microscopy systems that can comprehensively characterize 3D nanoscale alterations in nuclear architecture in carcinogenesis using clinically obtained routine formalin-fixed and paraffin-embedded tissue. Our optical microscopy systems include depth-resolved spatial-domain low-coherence quantitative phase microscopy (depth-resolved SL-QPM) and spectral-encoding of spatial frequency (SESF). We showed that depth-resolved SL-QPM detects structural changes at a sensitivity of 1 nm within a single cell nucleus, while SESF extracts the structural length-scale distribution at an accuracy of ~10-20 nm. Our extensive preliminary data have shown the promise of these optical markers to detect the presence of invasive cancer even from histologically normal cells from multiple tumor types and predict cancer progression risk. Now we propose to use these two optical microscopy systems together with state-of-the-art 3D super-resolution microscopy, to define a set of optical markers and the underlying nanoscale changes in nuclear architecture that are characteristic of each phase of tumorigenesis and identify those that detect "premalignant" changes. Then we will perform a clinical study to evaluate the accuracy of optical markers of nanoscale changes in nuclear architecture to predict breast cancer progression risk among women with pre-cancerous lesions (e.g., atypical hyperplasia (AH)) and pre-invasive cancer of ductal carcinoma in situ (DCIS) to avoid over- treatment. This project, if successful, will establish the alterations of nanoscale nuclear architecture in carcinogenesis, and have profound impact on both tumor biology research and clinical care. It will build a solid foundation for future use of optical markers of nanoscale changes in nuclear architecture as accurate prognostic markers to predict those women that are likely to progress into invasive cancer.