鼻咽癌患者T细胞受体库多样性特点及其临床意义研究

Nasopharyngeal Carcinoma (NPC) occurs with highest incidence in Southeastern China. Successful treatment of NPC, which depends largely on early detection, is the key to improve survival rate and life qualities of patients with NPC. Immune function of T cells in patients with NPC are closely related to the occurrence and development of tumor. The composition and function of T cells in NPC patients can be identified by the detection of TCR library diversity, which showed important potential reference value in tumor diagnosis and treatment. Our group makes the first attempt to carry out the research about clinical significance of TCR repertoire diversity, have established CDR3 spectratype analysis technology platform for detection of TCR repertoire diversity previously, and found a positively correlated between TCR repertoire diversity restore and tumor remission in patients with colorectal cancer after treatment. The project plan to make NPC as a disease breakthrough point, combination of the established CDR3 spectratype analysis technology and the latest amplicon rescued multiplex PCR as well as high-throughput sequencing technology, to analysis the TCR repertoire diversity and its correlation with therapeutic effect of five types of T-lymphocyte subsets in peripheral blood of 50 cases of patients with NPC, which provide basis for the application of TCR repertoire diversity in the relapse, metastasis and prognosis judgment of NPC. At the same time, find NPC specific TCR spectrum by analysis TCR of dominant T cell clones, to provide new molecular identification for the early diagnosis of NPC. This Project not only provides the basis for the application of TCR repertoire diversity in assistant clinical diagnosis and treatment of NPC, but also make a mode and platform for similar research in other T cells immune related diseases.

鼻咽癌是我国广东省高发的恶性肿瘤，早发现、早治疗是提高生存率的关键。鼻咽癌的发生发展与患者T细胞免疫功能密切相关。检测TCR库多样性，可了解T细胞的组成和功能，对于肿瘤诊断和治疗均有重要参考价值。课题组最早开展TCR库多样性及其临床意义研究。前期已建立检测TCR库多样性的CDR3谱型分析技术平台，并发现结直肠癌患者治疗后TCR库正常化与肿瘤缓解呈正相关。本项目拟以鼻咽癌作为切入点，利用已建立的CDR3谱型分析技术与最新的扩增子救援多重PCR和高通量测序技术相结合，分析50例鼻咽癌患者外周血5种T细胞亚群治疗前后多个阶段TCR库多样性及其与疗效相关性，为其应用于复发、转移和预后判断提供依据；同时，通过分析优势克隆TCR，找到疾病特征性TCR谱，为鼻咽癌早期诊断提供新的分子标识。项目不仅为TCR库多样性分析辅助鼻咽癌临床诊断和治疗提供依据，也为其他T细胞免疫相关疾病的同类研究提供模式和平台。

鼻咽癌是我国广东省高发的恶性肿瘤，早发现、早治疗是提高生存率的关键。鼻咽癌的发生发展与患者T细胞免疫功能密切相关。检测TCR库多样性，可了解T细胞的组成和功能，对于肿瘤诊断和治疗均有重要参考价值。. 本项目选取54例将接受常规放化疗的鼻咽癌患者作为研究对象；同时收集了33名健康人、39名同期住院的非鼻咽癌患者（鼻炎、鼻息肉等）外周血作为研究对照；建立标准化、稳定、自主创新的高通量全克隆TCR免疫组库检测技术平台（发明专利已公开），检测15例鼻咽癌患者癌与癌旁活检组织中的T细胞TCR免疫组库；检测全部献血者的外周血TCR免疫组库。采用生物信息学方法分析高通量测序数据，通过与健康人、疾病对照组TCR库对比，结合LASSO回归分析筛选出差异主成分TRBV基因，建立鼻咽癌特征性TCR 谱：随机选择20名NPC患者、19名疾病对照和17名健康对照作为训练集，剩余一半组成验证集。对训练集进行LASSO分析挑选出15个TRBV基因通过所得公式计算TRBV得分可区分NPC患者与对照。ROC曲线分析训练集的AUC值为0.96,验证集为0.741。对15位鼻咽癌患者癌与癌旁组织样本进行TCR免疫组库分析。因鼻咽解剖部位的特殊性，人与人之间存在高度异质性，故以癌旁组织为参考基线比较癌组织间的差异。结果发现癌组织TCR库多样性低于癌旁组织预示较差的预后。鼻咽癌患者外周血TCR库多样性显著高于健康对照和疾病对照组，且与不良预后正相关，该指标独立于TNM分期和EBV水平，TCR库多样性指数U/T与TNM分期和EBV水平结合可显著提升预测效能。. 项目以鼻咽癌作为切入点，建立了能区分鼻咽癌与非癌人群的鼻咽癌特征性TCR 谱。并将TCR免疫组库数据与患者临床资料进行关联性分析，探讨了TCR免疫组库分析在鼻咽癌诊断、预后中的应用价值。同时，还开展了TCR免疫组库分析在肝癌、乳腺癌、胃癌等其他肿瘤预后中的临床意义研究。项目不仅为TCR免疫组库分析辅助鼻咽癌临床诊断和预后提供依据，也为其他T细胞免疫相关疾病的同类研究提供模式和平台。