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

项目摘要 尽管取得了巨大的进展，但很大一部分癌症患者不会从最新的治疗中受益。 由于其肿瘤的特点，在免疫治疗发展方面取得了突破。t细胞受体 靶向肿瘤特异性新抗原的工程化T细胞（TCR T细胞）已经作为一种有希望的方法出现， 提高药效，降低毒性，克服获得性抗性。然而，高度个性化的性质 这种治疗方法仅仅依赖于识别特定癌症中表达的抗原， 相应的T细胞受体（TCR）使它们的发展特别具有挑战性。确定和 免疫原性新抗原靶标及其相关TCR的表征已被认为是免疫原性新抗原靶标的主要特征。 由于癌症中突变的多样性和缺乏高通量方法来识别 新抗原特异性TCR。因此，目前大多数临床发展都集中在肿瘤 这些突变在癌症患者中具有很高的流行率，使许多患者没有治疗选择。 为了增加临床相关目标和候选人的数量，从而增加合格患者的数量，Flexomics 正在开发下一代筛选平台，致力于高通量发现， 抗原特异性TCR的表征。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独特的平台，专为 筛选新抗原/T细胞相互作用。我们的专有平台旨在识别T细胞活化， 对100，000个T细胞在单细胞水平下暴露于特异性抗原的应答，同时捕获 以关键标记基因和全长TCR的表达形式的表型和基因型信息 序列鉴定基于我们令人鼓舞的可行性结果，我们建议展示可扩展性 我们的方法的功能相关性：（1）通过询问多个 抗原，（2）整合，简化和自动化我们的过程的所有不同组成部分，（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。在我们第二阶段的资助结束时， 我们将处于理想的位置，开始与外部客户、战略合作伙伴和投资者接触， 采用我们的平台并将生成的数据转化为治疗发展。

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.