Novel Approach to High-Throughput Identification and Characterization of Neoantigen-Specific T-Cell Receptors to Guide Immunotherapy Development.

项目总结 尽管取得了巨大的进步，但很大一部分癌症患者将不会从最新的 由于其肿瘤的特点，在免疫治疗方面的发展取得了突破。T细胞受体 靶向肿瘤特异性新抗原的工程化T细胞(TCRT细胞)已作为一种有希望的方法出现 提高疗效，降低毒性，克服获得性耐药性。然而，高度个性化的性质 仅依赖于识别特定癌症中表达的抗原的这种治疗方法 相应的T细胞受体(TCR)使得它们的开发尤其具有挑战性。身份识别和 免疫原性新抗原靶标及其相关的TCR的特征已被认为是主要的 由于癌症突变的多样性和缺乏高通量的鉴定方法而带来的挑战 新抗原特异性TCRs。因此，目前的大多数临床进展都集中在肿瘤上。 在癌症患者中发病率很高的突变，使许多患者没有治疗选择。 为了增加临床相关靶点和候选对象的数量，从而增加符合条件的患者池，灵活组学 正在开发下一代筛查平台，致力于高通量发现和 抗原特异性TCRs的特性。我们的方法结合了活细胞分析、黄金标准 免疫分析和最先进的单细胞基因组学在一个专门为 新抗原/T细胞相互作用的筛选。我们的专有平台旨在识别T细胞在 同时捕获100,000个T细胞对单细胞水平特异性抗原暴露的反应 关键标记基因和全长TCR表达形式的表型和基因信息 序列标识。基于我们前景看好的可行性结果，我们建议演示可伸缩性 和我们方法的功能相关性：(1)通过询问多个 抗原，(2)集成、简化和自动化我们过程的所有不同组件，(3)验证 我们的工作流程，并为100个选定的癌症靶点生成概念验证数据。在我们的第二阶段拨款结束时， 我们将处于与外部客户、战略合作伙伴和投资者接触的理想位置，以推动 采用我们的平台，并将产生的数据转化为治疗开发。

PROJECT SUMMARY Despite tremendous progress, a significant fraction of cancer patients will not benefit from the latest breakthroughs in immunotherapy development due to the characteristics of their tumor. T-cell Receptor engineered T-cells (TCR T-cells) targeting tumor specific neoantigens has arisen as a promising approach to improve efficacy, decrease toxicity and overcome acquired resistance. However, the highly personalized nature of such treatments that relies solely on the recognition of antigens expressed in specific cancers by corresponding T-Cell Receptors (TCRs) makes their development especially challenging. The identification and characterization of immunogenic neoantigen targets and their associated TCRs has been recognized as a major challenge due to the multiplicity of mutations in cancer and the lack of high-throughput methods to identify neoantigen-specific TCRs. Consequently, most current clinical developments have been focused on tumor mutations that have high prevalence among cancer patients, leaving many patients without therapeutic options. To increase the number of clinically relevant targets and candidates, thus the pool of eligible patients, Flexomics is developing a next generation of screening platform dedicated to the high-throughput discovery and characterization of antigen-specific TCRs. Our approach combines live cell analysis, gold-standard immunoassays and state-of-the-art single cell genomics in a unique platform specifically designed for the screening of neoantigen/T-cell interactions. Our proprietary platform is designed to identify T-cell activation in response to specific antigen exposure at single cell level for 100,000s T-cells while simultaneously capturing phenotypic and genotypic information in the form of expression of key marker genes and full-length TCR sequence identification. Building on our promising feasibility result, we propose to demonstrate the scalability and functional relevance of our method by: (1) increasing our screening capabilities by interrogating multiple antigens, (2) integrating, streamlining and automating all the different components of our process, (3) validate our workflow and generate proof-of-concept data for 100 select cancer targets. At the end of our Phase II grant, we will be ideally positioned to start engaging with external customers, strategic partners and investors to drive the adoption of our platform and the translation of the data generated into therapeutic developments.