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/摘要 尽管它们成功了，但免疫疗法无法以抗原依赖性的基础与T细胞接合，而是选择 基于其位置或表型，重新激活一大群特异性的T细胞。这样的标记 由于PD-1已被提倡描绘抗肿瘤T细胞，但也以认识非 - 同样，抑制肿瘤的T细胞以排气 我们和其他人也证明了还识别病毒和细菌抗原（即旁观者T细胞）。 MHC 多聚体提出了识别抗肿瘤T细胞的潜在替代方案，但受到前部需要的限制 识别抗原感兴趣的抗原。由于缺乏对抗原的了解，这种限制更加复杂 免疫原性和识别它们的T细胞。最近发现肿瘤突变的影响 Burnen（TMB）增强了我们对肿瘤抗原起源的掌握，Dospite TMB无法始终如一地 预测临床结果。这可能是最好由悬停在 〜1％。因此，我们对哪些T细胞在实体瘤中暴露了治疗性抗肿瘤电位的理解 仍然有问题。因此，对能够识别抗抗衡的公正方法有迫切需要 肿瘤T细胞反应，缺乏会大大阻碍实体瘤中免疫疗法的进展。 我们以前开发了附着（评估T细胞束缚在抗原I/II类组织相容性上），A 强大的微流体分析，该测定允许通过利用HLA/肽结合直接隔离抗肿瘤T细胞 T细胞受体（TCR）与匹配的肿瘤细胞的亲和力，事实上是“四聚体池”。在这里，我们建议 固定器的开发（通过电热增强恢复固定），一种微流体设备 能够在提高灵敏度，特异性和抗原特异性T细胞恢复的同时简化该测定法。 我们的中心假设是，T细胞识别可以在体外直接分离多种肿瘤抗原 T细胞的固定器，这些细胞携带的抗肿瘤细胞毒性潜力增加了 细胞。我们已经根据初步研究提出了这一假设 富含细胞毒性潜力增加的肿瘤浸润淋巴细胞（TILS）。提议的理由 研究是，通过开发T细胞固定器，应用电热流以附加将促进T 细胞/肿瘤细胞接触，并允许T细胞扫描多个MHC/抗原复合物以遇到它们 同源抗原。在AIM 1中，我们将设计和构建T细胞固定器并评估其恢复的能力 跨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。在AIM 2中，我们将在表型和 在功能上使用配对的人类衍生的Xenographtics和来自图标的肿瘤浸润淋巴细胞 队列。我们预计我们的工作将使我们能够证明T细胞固定器及其能力的多功能性 在体外改善抗肿瘤反应。总体而言，我们的工作将允许直接，准确和公正的身份证明 在数小时内而不是目前所需的几个月内，抗肿瘤T细胞的抗肿瘤T细胞。 随后研究该方法在实体瘤中的预测和治疗潜力的基础。