Investigation of Immune Modulation and Its Correlation to Response in Early Stage Lung Cancer Patients Receiving Neo-adjuvant Immunotherapy

肺癌是全球癌症相关死亡的主要原因。非小细胞肺癌(NSCLC)是其主要亚型。大多数患者是在疾病的晚期被诊断出来的，预后很差。最近癌症免疫学的突破表明，免疫检查点阻断(ICB)可以激活免疫反应。使用抗PD-1或抗PD-L1的ICB显示了良好的疗效，并与提高NSCLC的总体生存率有关。然而，只有15%-20%的患者显示出长期受益。ICB除影响抗肿瘤反应外，还可诱发炎性免疫相关不良反应(IrAEs)。肿瘤引流淋巴结在肿瘤的发生、发展和免疫中起着至关重要的作用。肺淋巴引流自肺内淋巴结(LN)至肺门和纵隔LN。ICB如何影响免疫细胞的激活，以及TDL、血液和肿瘤微环境中的免疫表型转变，目前还不完全清楚。詹姆斯胸科中心目前正在领导一项由研究人员发起的多中心第二阶段试验，以调查新辅助免疫疗法(NIT)与抗PD-L1制剂阿特唑珠单抗在可切除和未治疗的非小细胞肺癌患者中的有效性和安全性。作为这项临床试验的一部分，LN将在NIT之前和手术时进行采样。中心假设是，抗PD-L1的ICB将促进肿瘤特异性T细胞的激活和克隆性增殖，以及TDL、血液和肿瘤微环境的免疫表型转变，从而导致非小细胞肺癌的抗肿瘤免疫反应。为了验证这一假设，我们将进行三个特定目标的研究：AIM1使用治疗前后的纵隔和肺门LN样本(外科和EBUS-TBNA)定量分析NSCLC患者肿瘤浸润性和非肿瘤浸润性LN中的淋巴细胞数量，以评估免疫微环境的变化。AIM2定量分析治疗前后血样中循环细胞因子和循环细胞免疫表型，跟踪T细胞活化/克隆性增殖，并与结节免疫表型变化相关。AIM3将来自AIMs1和2的数据与对肿瘤浸润性免疫细胞群体的定量分析以及由合作机构获得的全面的肿瘤基因组学相关联，并与患者的反应和irAEs相关联。该项目将研究接受新辅助抗PD-L1治疗的早期非小细胞肺癌患者的免疫调节及其与疗效的相关性。这将有助于更好地了解肺癌的免疫反应机制，满足开发免疫相关生物标志物和优化肺癌免疫治疗的迫切需要。本研究还将从总体上阐明肺免疫调节机制和病理生理学，为以后研究和翻译肺部良恶性疾病的免疫相关研究奠定基础。

Lung cancer is the leading cause of cancer-related death worldwide. Non-Small Cell Lung Cancer (NSCLC) is its major subtype. Most patients are diagnosed at an advanced stage of disease and have a poor prognosis. Recent breakthroughs in cancer immunology suggest that immune-checkpoint blockade (ICB) can activate the immune response. ICB with anti-PD-1 or anti-PD-L1 shows promising efficacy and is associated with increased overall survival in NSCLC. However, only 15-20% of patients show a long-term benefit. Besides influencing the anti-tumor response, ICB could also induces inflammatory immune-related adverse events (irAEs). Tumor-draining lymph nodes (TDL) play a crucial role in cancer development, progression and immunity. The lung lymphatic drainage spans from intrapulmonary lymph nodes (LN) to hilar and mediastinal LN. How ICB impacts immune cell activation, as well as immunophenotype shifting in TDL, blood, and tumor microenvironment is not fully understood. The James Thoracic Center is currently leading a multicenter, investigator-initiated, Phase II trial to investigate the efficacy and safety of a neoadjuvant immunotherapy (NIT) with anti-PD-L1 agent atezolizumab in patients with resectable and untreated NSCLC. As part of this clinical trial, LN will be sampled prior to NIT and at the time of surgery. Blood and tumor samples will be taken as well.The central hypothesis is that ICB with anti-PD-L1 will promote tumor-specific T-cell activation and clonal expansion, as well as immunophenotype shifting in TDL, blood, and tumor microenvironment, leading to an anti-tumor immune response in NSCLC. To test this hypothesis, we will conduct research with three specific aims: Aim1 Quantitatively analyze lymphocyte populations in tumor-infiltrated and non-tumor infiltrated LN of NSCLC patients using pre- and post-treatment mediastinal and hilar LN samples (surgical and EBUS-TBNA) to evaluate for shifts in the immune microenvironment. Aim2 Quantitatively analyze circulating cytokines and circulating cell immunophenotypes in pre- and post-treatment blood samples, track T-cell activation/clonal expansion, and correlate with nodal immunophenotype changes. Aim3 Correlate data from Aims1 and 2 with quantitative analysis of tumor-infiltrating immune cell populations and comprehensive tumor genomics acquired by collaborating institutions, and with response and irAEs in patients. This project will investigate immune modulation and its correlation to response in early stage NSCLC patients receiving neo-adjuvant anti-PD-L1 therapy. This will help to better understand the immune response mechanism in lung cancer and to meet the crucial need for developing immune-related biomarkers and optimizing immunotherapy in lung cancer. This research will also shed light on lung immune modulation mechanism and pathophysiology in general, laying a foundation for future immunity related investigation and translation in both malignant and benign lung diseases.