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，也是全球癌症相关死亡的主要原因。 免疫疗法正在改变治疗机制，在肺癌方面进行的试验已经 显示出长期生存结果的改善。然而，现有的治疗方法并不是所有的都有效。 患者，强调需要更好地描述相关的复杂分子和免疫过程 与治疗反应和疾病进展有关。尽管T细胞一直是人类免疫缺陷病毒的主要目标 肺癌的免疫治疗，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)内其他免疫细胞活动的影响。肿瘤浸润性 一般情况下，B细胞在肿瘤进展中具有双重作用，但在非小细胞肺中 癌症(NSCLC)，特别是腺癌(LUAD)，特别是B细胞亚群 促进T细胞抗肿瘤反应，并对长期生存结果产生积极影响。关于以下角色的指示 最近还报道了LUAD中的特异性抗体(Ab)反应，包括肿瘤病灶的证据 抗原特异性反应。然而，抗肿瘤B细胞的特征及其功能特征具有 仅限于对相对较少数量的预选表面标记的研究 病人。然而，很明显，B细胞在LUAD肿瘤进展中的作用是不同的。 患者，并表明更全面和完整的特征的TI B细胞表型 抗肿瘤T细胞反应的变异和相关效应有很大的潜力告诉我们 了解疾病的进展，以及开发新的、潜在的个性化治疗方法。 这项提案的总体目标是定义与T细胞介导的T细胞相关的TI B细胞签名 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 首次全面分析肿瘤及其配对正常组织中淋巴细胞(B和T细胞)的技术 早期LUAD患者的大量队列，包括这种类型的数据与抗体谱系的第一次配对 测序作为询问LUAD B细胞关键功能抗体反应特征的一种手段。重要的是 建议的单细胞分析方法将利用已经确定与 LUAD的长期生存结果，使不同的功能B细胞类型及其与TI T细胞的相关性 可在疾病进展(目标1)的背景下具体确定。这些大规模的统计数据 相关性将得到验证，然后作为进行功能性体外培养和血清培养的框架- 基于分析以评估确定的预后良好的B细胞亚群是否具有不同的功能 与肿瘤抗原特异性抗体反应相关的表型，并对T细胞激活有不同的影响 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记物 对人类肺癌新疗法的发展至关重要。

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.