Super-Resolution Imaging of Higher-Order Heterochromatin Structure for Early Detection of Lung Carcinogenesis

项目摘要/摘要 肺癌是美国癌症相关死亡的主要原因。总体而言，被诊断为 早期肺癌的预后要好得多。吸烟是导致肺癌的主要原因。 目前，建议对目前和曾经吸烟的高危人群进行肺癌筛查。相比较 对于不吸烟的人，吸烟者患肺癌的风险几乎增加了30倍。不幸的是， 尽管付出了很大的努力，但肺癌的早期检测仍然不是传统方法的最佳选择。 假阳性率高或敏感度有限。提高了对早期事件的理解 潜在的吸烟相关肺癌的发生对于识别新的生物标记物和 早期发现和预防的目标。此外，检测肺癌早期事件的新方法 迫切需要以非侵入性、高灵敏度和高特异度的经济有效的方式进行开发 需要的。超分辨率显微镜的最新进展使光学显微镜领域发生了革命性的变化，并提供了 一种新的以纳米分辨率可视化在常规光线下不可见的分子结构的能力 显微镜。我们提出了一种创新的方法来适应超分辨率显微镜来改善早期的 肺癌的检测。我们的方法建立在我们最近的发现之上，即染色质折叠成为 在早期肺癌发生过程中逐渐碎片化，即使细胞在常规光线下仍显示正常 显微镜。我们的最终目标是检测这种纳米级染色质的错误折叠 痰，以提高肺癌的早期发现。在这个项目中，我们将首先建立被破坏的纳米尺度 利用致癌物诱导的小鼠肺模型，染色质折叠是肺癌发生的早期事件 腺癌以及现有注释良好的人肺组织标本(目标1)。此外，我们还将 评价纳米级染色质“错折叠”超分辨成像早期检测的可行性 利用现有的痰样本和匹兹堡肺筛查研究的数据来研究肺癌，这是一个社区- 基于现在和以前吸烟者的研究队列，用低剂量计算机断层扫描进行筛查，并进行跟踪 用于肺癌(目标2)。

PROJECT SUMMARY/ABSTRACT Lung cancer is the leading cause of cancer-related death in the United States. Overall, patients diagnosed with early-stage lung cancer have a much better prognosis. Cigarette smoking is the major cause of lung cancer and screening for lung cancer is currently recommended for high-risk current and former smokers. Compared with nonsmokers, smokers have an almost 30-fold increased risk of developing lung cancer. Unfortunately, despite significant efforts, early detection of lung cancer remains sub-optimal with conventional approaches suffering from high false-positive rates or limited sensitivity. Improved understanding of the early events underlying smoking-related lung cancer development is crucial to the identification of new biomarkers and targets for early detection and prevention. Further, new methods that detect those early events in lung cancer development in a non-invasive and cost-effective manner with high sensitivity and specificity are urgently needed. Recent advances in super-resolution microscopy revolutionize the field of optical microscopy and offer a new ability to visualize molecular structure at nanometer resolution that is invisible under a conventional light microscope. We propose an innovative approach to adapt super-resolution microscopy to improve the early detection of lung cancer. Our approach is built upon our recent discovery that chromatin folding becomes gradually fragmented in early lung carcinogenesis, even when cells still appear normal under conventional light microscope. Our ultimate goal is to detect such nanoscale chromatin “misfolding” in bronchial cells present in sputum to improve early detection of lung cancer. In this project, we will first establish disrupted nanoscale chromatin folding as an early event in lung carcinogenesis utilizing a mouse model of carcinogen-induced lung adenocarcinoma as well as existing well-annotated human lung tissue specimens (Aim 1). In addition, we will evaluate the feasibility of super-resolution imaging of nanoscale chromatin “misfolding” for early detection of lung cancer using existing sputum samples and data from the Pittsburgh Lung Screening Study, a community- based research cohort of current and ex-smokers, screened with low-dose computed tomography and followed for lung cancer (Aim 2).