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

项目总结/摘要 肺癌是美国癌症相关死亡的主要原因。总的来说，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 目前建议对高危的吸烟者和曾经吸烟的人进行肺癌筛查。相比 与不吸烟者相比，吸烟者患肺癌的风险增加了近30倍。不幸的是， 尽管做出了重大努力，但传统方法对肺癌的早期检测仍不理想 遭受高假阳性率或有限的敏感性。加深对早期事件的了解 潜在的吸烟相关的肺癌发展对于识别新的生物标志物至关重要， 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和成本有效的方式开发具有高灵敏度和特异性的 needed.超分辨率显微镜的最新进展彻底改变了光学显微镜领域， 一种以纳米分辨率可视化分子结构的新能力，在传统光线下是不可见的 显微镜我们提出了一种创新的方法，以适应超分辨率显微镜，以改善早期 肺癌的诊断我们的方法是建立在我们最近的发现，染色质折叠成为 在早期肺癌发生过程中逐渐碎裂，即使在常规光下细胞仍显示正常 显微镜我们的最终目标是检测存在于支气管细胞中的这种纳米级染色质“错误折叠”， 痰，以提高肺癌的早期发现。在这个项目中，我们将首先建立中断纳米 染色质折叠作为肺癌发生的早期事件，使用致癌物诱导的肺的小鼠模型 腺癌以及现有的注释良好的人肺组织标本（目标1）。此外，我们将 评估纳米级染色质“错误折叠”的超分辨率成像用于早期检测的可行性。 肺癌使用现有的痰样本和匹兹堡肺筛查研究的数据， 基于研究队列的当前和既往吸烟者，采用低剂量计算机断层扫描进行筛查，并随访 肺癌（Aim 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).