Unbiased electrothermal flow-enhanced identification of antigen-specific T cells in lung cancer

无偏电热流增强肺癌抗原特异性 T 细胞的鉴定

• 批准号: 10723218
• 负责人: Peter B Lillehoj
• 金额: $ 41.28万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10723218
• 关键词: Acceleration; Affinity; Anterior; Antigen-Antibody Reactions; Antigens; Antitumor Response; Avidity; Bacterial Antigens; Binding; Biological Assay; Biological Markers; Cell Communication; Cell Separation; Cells; Coculture Techniques; Complex; Comprehension; Development; Devices; Epitopes; Exhibits; Foundations; Geometry; Goals; Histocompatibility; Histocompatibility Antigens Class I; Hour; Human; Immunotherapy; In Vitro; Kinetics; Knowledge; Location; Malignant neoplasm of lung; Methods; Microfluidic Microchips; Microfluidics; Modeling; Mutation; OX40; Outcome; Peptides; Performance; Peripheral Blood Mononuclear Cell; Phenotype; Procedures; Property; Recovery; Research; Scanning; Sensitivity and Specificity; Solid Neoplasm; Specificity; T cell response; T-Cell Receptor; T-Lymphocyte; Technology; Testing; Therapeutic; Tumor Antigens; Tumor-Infiltrating Lymphocytes; Validation; Viral Antigens; Work; antigen binding; antigen-specific T cells; biomarker identification; cancer cell; clinical application; cohort; comparative; cost; cytotoxic; cytotoxicity; design and construction; experimental study; grasp; human model; immunogenic; improved; interest; mouse model; neoantigens; neoplastic cell; novel; particle; patient derived xenograft model; predict clinical outcome; programmed cell death protein 1; programs; receptor binding; shear stress; success; transcriptome sequencing; tumor

Project Abstract/Summary Despite their success, immunotherapies fail to engage T cells on an antigen-dependent basis rather opting to reactivate a broad spectrum of T cells of unknown specificity based on their location or phenotype. Markers such as PD-1 have been posited to delineate anti-tumor T cells but are also expressed on those recognizing non- tumor antigens. Likewise, tumor-resident T cells are surmised to exhibit tumor antigen recognition though they have been shown by us and others to also recognize viral and bacterial antigens (ie. bystander T cells). MHC multimers present a potent alternative to identify anti-tumor T cells but are limited by a need for anterior identification of the antigen(s) of interest. This limitation is compounded by a lack of understanding of the antigens that are immunogenic and the T cells that recognize them. Recent discoveries into the impact of tumor mutational burden (TMB) have bolstered our grasp of the origins of tumor antigens, despite TMB failing to consistently predict clinical outcomes. This is perhaps best epitomized by the dismal antigen validation rates which hover at ~1%. Hence, our understanding of which T cells exhibit therapeutic anti-tumor potential in solid tumors remains problematic. Therefore, there is a critical need for unbiased approaches capable of identifying anti- tumor T cell responses, a lack of which will substantially impede the progress of immunotherapies in solid tumors. We previously developed ATTACH (Assessment of T cells Tethered to Antigen Class I/II Histocompatibility), a powerful microfluidics assay which allows direct isolation of anti-tumor T cells by leveraging HLA/peptide binding avidity of T cell receptors (TCR) to matched tumor cells serving as de facto “tetramer pools”. Here, we propose the development of an ATTACHER (ATTACH via Electrothermal flow-enhanced Recovery), a microfluidic device capable of streamlining this assay while increasing sensitivity, specificity, and antigen-specific T cell recovery. Our central hypothesis is that T cells recognizing multiple tumor antigens can be directly isolated in vitro using a T cell ATTACHER, and that these cells harbor increased anti-tumor cytotoxic potential compared to bulk T cells. We have formulated this hypothesis on the basis of preliminary studies outlining the ability of ATTACH to enrich for tumor-infiltrating lymphocytes (TILs) with increased cytotoxic potential. The rationale for the proposed research is that applying electrothermal flow to ATTACH via development of a T cell ATTACHER will promote T cell/tumor cell contact and allow T cells to scan multiple MHC/antigen complexes in order to encounter their cognate antigen. In Aim 1, we will design and construct the T cell ATTACHER and evaluate its ability to recover antigen-specific T cells across a set of 20 different human antigens and TCRs of different affinities developed in our lab. In Aim 2, we will characterize TILs isolated using the T cell ATTACHER both phenotypically and functionally using paired human patient-derived xenografts and tumor-infiltrating lymphocytes from the ICON cohort. We anticipate our work will allow us to demonstrate the versatility of the T cell ATTACHER and its ability to improve anti-tumor responses in vitro. Overall, our work will allow direct, accurate and unbiased identification of anti-tumor T cells in solid tumors within hours rather than months as is currently required, laying the foundation for subsequent studies into the predictive and therapeutic potential of this method in solid tumors.

项目摘要/总结 尽管免疫疗法取得了成功，但它未能在抗原依赖性基础上吸引 T 细胞，而是选择 根据其位置或表型等标记重新激活未知特异性的广谱 T 细胞。 因为 PD-1 已被定位为描绘抗肿瘤 T 细胞，但也表达在那些识别非肿瘤 T 细胞上。 同样，肿瘤驻留 T 细胞据推测也表现出肿瘤抗原识别能力。 我们和其他人已经证明它也能识别病毒和细菌抗原（即旁观者 T 细胞）。 多聚体提供了一种有效的替代方法来识别抗肿瘤 T 细胞，但受到前壁需求的限制 由于缺乏对抗原的了解，这一限制变得更加复杂。 具有免疫原性的 T 细胞以及识别它们的 T 细胞最近发现了肿瘤突变的影响。 负担（TMB）增强了我们对肿瘤抗原起源的掌握，尽管TMB未能始终如一地 预测临床结果可能是徘徊在令人沮丧的抗原验证率的最佳体现。 因此，我们了解哪些 T 细胞在实体瘤中具有治疗性抗肿瘤潜力。 因此，迫切需要能够识别反病毒的公正方法。 肿瘤 T 细胞反应，缺乏这种反应将严重阻碍实体瘤免疫治疗的进展。 我们之前开发了 ATTACH（评估与 I/II 类抗原相连的 T 细胞组织相容性） 强大的微流体检测，可利用 HLA/肽结合直接分离抗肿瘤 T 细胞 T 细胞受体 (TCR) 与匹配的肿瘤细胞的亲和力充当事实上的“四聚体池”。 开发一种微流体装置 ATTACHER（通过电热流增强恢复连接） 能够简化该测定，同时提高灵敏度、特异性和抗原特异性 T 细胞回收率。 我们的中心假设是识别多种肿瘤抗原的 T 细胞可以直接在体外分离 T 细胞附着物，并且与散装 T 细胞相比，这些细胞具有更高的抗肿瘤细胞毒性潜力 我们根据概述 ATTACH 能力的初步研究制定了这一假设。 丰富具有增加细胞毒性潜力的肿瘤浸润淋巴细胞（TIL）。 研究表明，通过开发 T 细胞 ATTACHER 将电热流应用于 ATTACH 将促进 T 细胞/肿瘤细胞接触，并允许 T 细胞扫描多个 MHC/抗原复合物，以便遇到它们 在目标1中，我们将设计和构建T细胞ATTACHER并评估其恢复能力。 跨越 20 种不同人类抗原和不同亲和力的 TCR 的抗原特异性 T 细胞 在我们的实验室的目标 2 中，我们将表征使用 T 细胞附着物分离的 TIL 的表型和特征。 功能性地使用来自 ICON 的配对人类患者异种移植物和肿瘤浸润淋巴细胞 我们预计我们的工作将使我们能够展示 T 细胞附着物的多功能性及其能力。 总体而言，我们的工作将实现直接、准确和公正的识别。 实体瘤中的抗肿瘤 T 细胞可在数小时内（而不是目前所需的数月内）产生，从而奠定了 为后续研究该方法在实体瘤中的预测和治疗潜力奠定了基础。