Imaging nanoscale chromatin folding in early carcinogenesis

项目摘要 这项应用是为了评估超分辨率显微镜系统对图像破坏的潜力 纳米级染色质折叠作为癌变的早期事件并探索其改善癌症的潜力 风险分层。染色质结构异常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的历史。然而，由于衍射有限的分辨率 常规光学显微镜只能观察到微尺度的结构异常。因此，细胞 经历恶变的早期阶段通常看起来是正常的。这种图像分辨率限制 损害了我们准确地对前驱病变进行风险分层或区分好斗和懒惰的能力 表格。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 染色质结构分辨率为~20-30 nm。我们集团最近提高了吞吐量和 随机光学重建显微镜(STORM)超分辨率显微镜的稳健性 在最广泛使用的临床样本上实现了染色质折叠的稳健成像。建立在我们的 初步研究显示，在早期，纳米级染色质折叠显著并逐渐中断 在致癌方面，这个项目将首先进一步确认伴随着染色质折叠的破坏 并确定它们的分子特征和功能后果。第二，我们将 优化超分辨率成像系统、样品制备和图像分析的工作流程，以实现 临床组织样本中纳米级染色质折叠的高效和重复性分析。我们还将 证实我们在各种大肠前驱病变患者中发现染色质折叠中断和 癌症。第三，我们将评估成像纳米级染色质折叠在患者中的潜力 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图谱 打乱了早期致癌过程中的纳米级染色质折叠，为风险分层开辟了一条新的途径， 促进制定和评估新的预防战略。

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.