Exosomal Thomsen-Friedenreich glycoantigen as a new biomarker for lung cancer screening and early detection

项目摘要 尽管近年来肺癌死亡率有所改善，但它仍然是全球主要的癌症杀手。低 高风险患者的剂量计算机断层扫描（CT）扫描筛查显示， 然而，存活率，它携带> 95%的假阳性率，随后的风险是不必要的侵入性诊断 活组织检查和反复辐射已经开发了液体活检测定来克服这些问题。 挑战，并提高CT筛查肺结节的诊断准确性。在许多生物标志物中， 肿瘤来源的外来体（TEX）已经成为有效的癌症生物标志物。我们发现了一种新的 外泌体标记物，T-Ag-α; Gal β 1 - 3GalNAc α，作为潜在的 肺癌早期检测的生物标志物。TF-Ag-α是一个独特的靶点，表达于约84%的细胞表面。 肺癌（包括非小细胞肺癌和小细胞肺癌）和其它癌（如乳腺癌、结肠癌 癌症等），但不在正常组织上。目前，关于TEX TF-Ag-α及其在细胞凋亡中的作用尚未见报道。 筛查和早期发现肺癌和其他癌症。在这个项目中，我们选择肺癌作为 疾病模型，目的是证明TEX TF-Ag-α在癌症液体活检中的临床实用性。转型 我们的发现应用于临床，我们已经开发了一种具有高度特异性的单克隆抗体JAA-F11， 和基于表面等离子体共振（SPR）的液体活检测定，以测量TF-Ag-α的水平。 外泌体TF-Ag-α。我们首次证明了外泌体携带TF-Ag-α。更 重要的是，我们成功地检测了低至10 μ L血清样品中的外泌体TF-Ag-α， 和晚期非小细胞肺癌（NSCLC）和小细胞肺癌（SCLC）患者，但几乎没有信号 与正常对照组相比。基于我们有希望的初步数据，我们假设外泌体TF-Ag-α是一种 肺癌筛查和早期检测的有效生物标志物。在这个项目中，我们建议（1）进一步发展 并全面表征其传感性能，包括分析 灵敏度（检测限、定量限）、特异性、线性范围和重复性;（2）证明 外泌体TF-Ag-α在肺癌筛查和早期检测中的诊断价值 来自明确定义的患者队列的外来体和血清样品，包括肺癌患者（NSCLC患者和非小细胞肺癌患者）。 和SCLC），具有肺癌高风险的良性肺结节的个体和没有 肺结节（共n = 200）。我们的目标是开发一种有效的癌症特异性生物标志物和一种准确的 液体活组织检查补充低剂量CT筛查中检出的肺结节诊断 提高低剂量CT的准确性。

PROJECT SUMMARY Although lung cancer mortality has improved in recent years, it remains the leading cancer killer worldwide. Low dose computed tomography (CT) scan screening in high-risk patients has shown improved cancer-specific survival, however, it carries >95% of false-positive rate, a subsequent risk of unnecessary invasive diagnostic biopsy, and repeated radiation exposure. Liquid biopsy assays have been developed to overcome these challenges and to enhance the diagnostic accuracy of CT-screened lung nodules. Among many biomarkers, tumor-derived exosomes (TEXs) have emerged as potent cancer biomarkers. We have discovered a new exosomal marker, Thomsen-Friedenreich glycoantigen (TF-Ag-α; Galβ1-3GalNAc alpha), as a potential biomarker for lung cancer early detection. TF-Ag-α is a unique target expressed on the cell surface of about 84% lung cancers (both nonsmall cell and small cell lung cancer) and other carcinomas (such as breast cancer, colon cancer, etc), but not on normal tissues. Currently there are no reports about TEX TF-Ag-α and its roles in the screening and early detection of lung cancer and other cancers. In this project, we select lung cancer as the disease model and aim to demonstrate the clinical utility of TEX TF-Ag-α in cancer liquid biopsy. To transform our discovery to clinical applications, we have developed a monoclonal antibody, JAA-F11, with high specificity to TF-Ag-α and a surface plasmon resonance (SPR)-based liquid biopsy assay to measure the levels of exosomal TF-Ag-α in blood. We have demonstrated, for the first time, that exosomes carry TF-Ag-α. More importantly, we successfully detected exosomal TF-Ag-α in as low as 10uL serum samples from both early stage and late stage nonsmall cell lung cancer (NSCLC) and small cell lung cancer (SCLC) patients, but little signals from those of normal controls. Based on our promising pilot data, we hypothesize that exosomal TF-Ag-α is a potent biomarker for lung cancer screening and early detection. In this project, we propose to (1) further develop the liquid biopsy assay and comprehensively characterize its sensing performances including analytical sensitivity (limit of detection, limit of quantitation), specificity, linear range, and repeatability; (2) demonstrate the diagnostic values of exosomal TF-Ag-α in lung cancer screening and early detection using both cell-derived exosomes and serum samples from well-defined cohorts of patients including lung cancer patients (both NSCLC and SCLC), individuals with benign pulmonary nodules at high risk of lung cancer and normal controls with no pulmonary nodules (total n=200). We aim to develop an effective cancer specific biomarker and an accurate liquid biopsy assay to complement the diagnosis of pulmonary nodules detected during low dose CT screening and to enhance the accuracy of low dose CT.