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

描述(申请人提供)：这项拟议的项目是为了回答C组提出的挑衅性问题2：如何利用肿瘤的物理属性，如细胞的电、光学或机械属性，来提供更早或更可靠的癌症检测、诊断、预后或药物反应或肿瘤复发的监测？我们将研究癌变过程中细胞核的光学性质及其相关的纳米尺度结构变化，并确定其准确性，以提供更早和更准确的乳腺癌诊断和预后。我们假设，核结构的纳米尺度变化发生在癌变的早期，而对核结构纳米尺度变化的光学标记的测量可以作为一种经济有效和准确的工具，用于早期和更准确的癌症诊断和预后。我们团队开发了一套光学显微镜系统，使用临床获得的常规福尔马林固定和石蜡包埋组织，可以全面表征致癌过程中核结构的3D纳米级变化。我们的光学显微镜系统包括深度分辨空域低相干定量位相显微镜(深度分辨SL-QPM)和空间频率光谱编码(SESF)。我们发现，深度分辨的SL-QPM以1 nm的灵敏度检测单个细胞核内的结构变化，而SESF以~10-20 nm的精度提取结构的长度-尺度分布。我们广泛的初步数据显示，这些光学标记物有望检测浸润性癌症的存在，甚至从多种肿瘤类型的组织学正常细胞中检测，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超分辨率显微镜结合使用，以定义一组光学标记和核结构中潜在的纳米级变化，这些变化是肿瘤发生的每个阶段的特征，并确定那些检测到“癌前”变化的。然后，我们将进行一项临床研究，以评估核结构纳米级变化的光学标记物预测患有癌前病变(例如，不典型增生(AH)和导管原位癌(DCIS)前浸润癌)的女性乳腺癌进展风险的准确性，以避免过度治疗。该项目如果成功，将为肿瘤发生过程中纳米尺度核结构的改变奠定基础，并将对肿瘤生物学研究和临床治疗产生深远影响。这将为未来使用纳米级核结构变化的光学标记作为准确的预后标记奠定坚实的基础，以预测那些可能进展为浸润性癌症的女性。

DESCRIPTION (provided by applicant): 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 markrs of nanoscale changes in nuclear architecture as accurate prognostic markers to predict those women that are likely to progress into invasive cancer.