Imaging nanoscale chromatin folding in early carcinogenesis

项目摘要 本申请是为了评估潜在的超分辨率显微镜系统的图像中断 纳米级染色质折叠作为癌发生的早期事件，并探索其改善癌症的潜力 危险分层异常染色质结构是肿瘤细胞最普遍的特征之一， 用于临床癌症诊断已有两个世纪。然而，由于衍射极限分辨率 传统的光学显微镜，只能观察到微观结构异常。因此，细胞 经历恶性转化的早期阶段通常表现正常。图像分辨率的这种限制 已经损害了我们准确地对前驱病变进行风险分层或区分侵袭性和惰性的能力， forms.超分辨率荧光纳米显微镜的最新进展使我们能够在分子水平上成像 染色质结构的分辨率低至约20-30 nm。我们小组最近提高了吞吐量， 基于随机光学重构显微镜（STORM）的超分辨率显微镜的鲁棒性， 能够在最广泛使用的临床样品上对染色质折叠进行稳健成像。建立在我们的 初步研究显示，在早期， 致癌作用，该项目将首先进一步证实，伴随着破坏染色质折叠 致癌作用并确定其分子特征和功能后果。二是 优化超分辨率成像系统、样品制备和图像分析的工作流程， 临床组织样品中纳米级染色质折叠的高效和可重复分析。我们还将 证实了我们在各种结直肠前病变患者中染色质折叠破坏的发现， 癌第三，我们将评估成像纳米染色质折叠的潜力， 结直肠腺瘤性息肉。这项研究将建立科学基础和潜在的分子概况， 在早期癌变过程中破坏了纳米级染色质折叠，为风险分层开辟了新途径， 促进制定和评价新的预防战略。

PROJECT ABSTRACT This application is to evaluate the potential of a super-resolution microscopy system to image disrupted nanoscale chromatin folding as an early event in carcinogenesis and explore its potential to improve cancer risk stratification. Abnormal chromatin structure is among the most universal characteristics of tumor cells and has been used for clinical cancer diagnosis for two centuries. However, due to the diffraction-limited resolution of conventional light microscopy, only microscale structural abnormalities can be observed. As a result, cells undergoing early stages of malignant transformation often appear normal. Such limitation in image resolution has compromised our ability to accurately risk-stratify precursor lesions or distinguish aggressive from indolent forms. Recent advances in super-resolution fluorescence nanoscopy now enable us to image molecular-level chromatin structure down to a resolution of ~20-30 nm. Our group recently improved the throughput and robustness in stochastic optical reconstruction microscopy (STORM)-based super-resolution microscopy and enabled robust imaging of chromatin folding on the most widely used clinical samples. Built upon our preliminary studies that revealed a significant and gradual disruption of nanoscale chromatin folding in early carcinogenesis, this project will first further confirm the disrupted chromatin folding that accompanies carcinogenesis and identify their molecular characteristics and functional consequences. Second, we will optimize the workflow of super-resolution imaging system, sample preparation and image analysis to enable efficient and reproducible analysis of nanoscale chromatin folding in clinical tissue samples. We will also validate our finding of disrupted chromatin folding in patients with various colorectal precursor lesions and cancer. Third, we will evaluate the potential of imaging nanoscale chromatin folding to in patients with colorectal adenomatous polyps. This study will establish the scientific basis and underlying molecular profile of disrupted nanoscale chromatin folding in early carcinogenesis, opening a new avenue for risk stratification, facilitating the development and evaluation of new preventive strategies.