自身免疫抗体构象与非小细胞肺癌预后相关性的研究

Non-small cell lung cancer (NSCLC) has a very dismal prognosis with an overall 5-year survival rate less than 15%. The traditional histopathological examination and TNM staging system are unable to accurately predict the therapeutic outcomes. Many NSCLC patients who have potentials of recurrence and metastasis could not be detected in a timely manner, thus, have lost the chance for effective treatments. In this study, we will take the advantage of immune surveillance to identify autoantibody profiles for assessing NSCLC development and therapeutic outcomes using phage-display and protein microarrays. We have previously achieved 90% sensitivity and 95% specificity in detection of NSCLC using a combined 5 autoantibodies that were identified using the same approach. These 5 autoantibody biomarkers have been pending for US patent. Moreover, our preliminary result showed that the same 5 autoantibodies are also capable to separate good and bad prognosis in 5-year survivals (p < 0.001) using Kaplan-Meier analysis. In this project, we will screen and validate autoantibody profiles for good prognostic biomarkers using a large number of banked NSCLC serum samples with detailed follow-up data. Particularly we will develop statistical classifiers that are capable to accurately predict prognosis either for 5-year survivals for post-operation patients or 2-year survivals for mid-late stage NSCLC patients. Hence the prognostic status would provide useful information in terms of patients' treatments and managements.

非小细胞肺癌（NSCLC）患者总体5年生存率低于15%。传统的病理学检查和TNM分期尚不能对NSCLC的预后进行准确地判断。为了开发新型准确的NSCLC预后分子标志物，本项目根据免疫监视原理，结合噬菌体展示和蛋白芯片技术，拟从患者血清中筛选出与NSCLC预后评价相关的自身免疫抗体，利用多种自身抗体构象来对患者病情发展及治疗效果进行评估和预测。申请人已运用此类方法成功筛选到了5种NSCLC早期诊断的自身抗体，联合诊断的敏感性达到90%特异性95%，该成果已申报了美国专利。预试验发现，这5种自身抗体联合检测还能对手术后NSCLC患者5年生存率有较准确预测（p<0.001）。在本项目中，我们将利用大量已存的NSCLC患者血清及随访结果，筛选出一组自身免疫抗体标志物，能够较为准确的对NSCLC术后病人5年或晚期病人2年生存率及疗效进行判断，使得具有潜在复发危险的患者能够得到及时的诊断和有效的治疗。

非小细胞肺癌（NSCLC）患者总体5年生存率低于15%。传统的病理学检查和TNM分期尚不能对NSCLC的预后进行准确地判断。为了开发新型准确的NSCLC预后分子标志物，本项目根据免疫监视原理，将噬菌体展示蛋白质分析技术与新兴的后基因组蛋白芯片研究手段相结合，应用于新型NSCLC 自身免疫抗体的筛选和预后分析研究，建立了一系列NSCLC 患者预后监测标志物体系。通过构建高丰度NSCLC组织cDNA T7噬菌体展示文库的构建，并利用NSCLC早中期（Ia-III a）和晚期（IIIb-IV）患者预后不良混合血清分别与相对应肿瘤分期的预后良好混合血清进行文库淘洗，筛选出了NSCLC预后特异性噬菌体克隆，并利用大量预后良好与不良的病人血清样本对预后特异性噬菌体克隆进行统计学验证，最终获得了9中特异性的自身免疫抗体标志物，其联合预后预测结果在对200份双盲血清样本的检测中，对早中期（适合手术治疗）评估准确度为86%，对于晚期（非手术治疗）NSCLC 患者预后评估的敏感性为94.3%，特异性为90.4%。为了进一步提高预后评估的准确性，课题组还对NSCLC血清中DNA甲基化、白细胞端粒长度在诊断及预后中的应用进行了研究。研究发现RUNX3、RASSF1A、3-OST-2、DAPK、PTPRO基因甲基化检出率明显高于正常人，联合检测可能有助于NSCLC早期诊断及其预后评估。本研究利用多种肿瘤标志物对NSCLC的复发转移进行联合监测，开发出了可运用于对病情发展及治疗效果进行评估的新型肿瘤标志物，为今后临床上对NSCLC的诊断及预后评估提供了一种新方法。