Defining tumor-infiltrating B cell signatures associated with T cell mediated antitumor immunity in human lung adenocarcinoma

肺癌是最常见的恶性肿瘤，是全球癌症相关死亡的主要原因。 免疫疗法正在改变治疗方案，在肺癌中进行的试验已经 在长期生存结果方面有所改善。然而，现有的治疗方法并不是对所有人都有效。 患者，强调需要更好地表征相关的复杂分子和免疫过程 与治疗反应和疾病进展有关。虽然T细胞一直是主要的目标， 随着T细胞介导的抗肿瘤免疫在肺癌中的应用，人们越来越认识到T细胞介导的抗肿瘤免疫可以 受肿瘤微环境（TME）内其他免疫细胞的影响。肿瘤浸润 (TI)B细胞已被证明在肿瘤进展中具有双重作用，但在非小细胞肺癌中， 癌症（NSCLC），特别是腺癌（LUAD），特别是B细胞亚群已经显示出对两种癌症都有 促进T细胞抗肿瘤反应，并对长期生存结果产生积极影响。作用的指示 最近也报道了LUAD中的特异性抗体（Ab）反应，包括肿瘤集中的证据 抗原特异性反应。然而，抗肿瘤B细胞及其功能特征的表征已经被证明是有效的。 在相对少量的研究中，仅对少量预先选择的表面标记进行了研究， 患者尽管如此，很明显B细胞介导的LUAD肿瘤进展中的作用是可变的， 患者，并表明一个更全面和综合的表征TI B细胞表型 变异及其对抗肿瘤T细胞应答的相关影响具有很大的潜力，可以为我们提供信息。 了解疾病进展，以及开发新的，潜在的个性化治疗。 该提案的总体目标是定义与T细胞介导的免疫应答相关的TI B细胞特征。 人肺癌的抗肿瘤反应。为了实现这一目标，我们将利用几个尖端技术， 首先全面分析肿瘤和配对正常组织中淋巴细胞（B和T细胞）的技术， 早期LUAD患者的大型队列，包括此类数据与Ab库的首次配对 测序作为询问LUAD B细胞中关键功能性Ab应答特征的手段。重要的是， 所提出的单细胞分析方法将利用已经确定与以下疾病相关的B细胞标志物： 长期LUAD生存结果，因此不同的功能性B细胞谱及其与TI T细胞的关联 可以在疾病进展的背景下具体确定（目标1）。这些大型统计 协会将得到验证，然后作为一个框架进行功能性体外培养和血清- 基于分析来评估所鉴定的免疫学上有利的B细胞亚群是否具有不同的功能 表型，与肿瘤抗原特异性抗体反应相关，并对T细胞活化有不同的影响 和效应器功能（目标2）。该项目的成功完成将促进新的发现和生物标志物 这对于开发人类肺癌的新疗法至关重要。

Lung cancers are the most common malignancies and are the leading cause of cancer-related deaths worldwide. Immunotherapies are transforming treatment regimes, and trials conducted in lung cancer have already demonstrated improvements in long-term survival outcomes. However, existing therapies are not effective in all patients, stressing a need to better characterize the complex molecular and immunological processes associated with treatment responsiveness and disease progression. Although T cells have been the primary target of immunotherapies in lung cancer, there is a growing appreciation that T cell-mediated anti-tumor immunity can be influenced by actions of other immune cells within the tumor microenvironment (TME). Tumor-infiltrating (TI) B cells have been shown to have dual roles in tumor progression generally, but in non-small cell lung cancers (NSCLC), specifically adenocarcinomas (LUAD), particular B cell subsets have been shown to both promote T cell anti-tumor responses and positively impact long-term survival outcomes. Indications of a role for specific antibody (Ab) responses in LUAD have also recently been reported, including evidence of focused tumor antigen-specific responses. However, characterizations of anti-tumor B cells and their functional signatures have been limited by the study of only a small number of pre-selected surface markers in relatively small numbers of patients. Nonetheless, it is evident that B cell-mediated roles in LUAD tumor progression are variable among patients, and indicates that a more comprehensive and integrated characterization of TI B cell phenotypic variation and the associated effects on the anti-tumor T cell response has great potential to inform our understanding of disease progression, as well as the development of novel, potentially personalized, therapies. The overall goal of this proposal is to define TI B cell signatures that are associated with the T cell-mediated anti-tumor response in human lung cancer. To achieve this goal, we will leverage several cutting-edge techniques to first comprehensively profile lymphocytes (B and T cells) in tumor and paired normal tissues from a large cohort of early-stage LUAD patients, including the first pairing of this type of data with Ab repertoire sequencing as a means to interrogate critical functional Ab response signatures in LUAD B cells. Importantly, the single-cell profiling approach proposed will leverage B cell markers already determined to be associated with long-term LUAD survival outcomes, so that distinct functional B cell profiles and their associations with TI T cells can be identified specifically in the context of disease progression (Aim 1). These large-scale statistical associations will be validated and then serve as a framework for conducting functional in vitro culture and serum- based analyses to assess whether prognostically favorable B cell subsets identified have distinct functional phenotypes, associate with tumor antigen-specific Ab responses, and have differential effects on T cell activation and effector function (Aim 2). Successful completion of this project will facilitate novel discovery and biomarkers critical for the development of novel therapeutics in human lung cancer.