Nano and biomolecular engineered technologies for neoantigen-specific T cell capture and characterization

用于新抗原特异性 T 细胞捕获和表征的纳米和生物分子工程技术

• 批准号: 10297588
• 负责人: James R. Heath
• 金额: $ 59.56万
• 依托单位: INSTITUTE FOR SYSTEMS BIOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-16 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10297588
• 关键词: Address; Algorithms; Antigen Presentation; Antigens; Antitumor Response; Artificial nanoparticles; Binding; Blood; Bypass; CCNE1 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; COVID-19 patient; Cells; Clinic; Clinical; Clone Cells; Community Clinical Oncology Program; Cost Savings; Data; Data Set; Databases; Disadvantaged; Engineering; Exhibits; Funding; Genes; Genetic; Grant; Haplotypes; Heart; Histocompatibility Antigens Class II; Immunooncology; Immunotherapy; Infusion procedures; KRAS2 gene; Length; Libraries; MHC Class II Genes; MHC antigen; Malignant neoplasm of ovary; Methodology; Methods; Mutate; Mutation; Nanotechnology; Oncogenes; Patients; Peptides; Peripheral Blood Mononuclear Cell; Population; Preparation; Protein Engineering; Proteins; Protocols documentation; Reagent; Reporting; Resources; Specificity; T cell response; T cell therapy; T-Cell Receptor; T-Cell Receptor Genes; T-Lymphocyte; T-Lymphocyte Subsets; TP53 gene; Technology; Therapeutic; Time; Tumor Antigens; Validation; Work; antigen-specific T cells; antitumor effect; base; cancer cell; cancer immunotherapy; colon cancer patients; cost; design; design and construction; innovation; invention; microchip; mutant; nano; neoantigens; pandemic disease; prediction algorithm; tool; tumor

Project summary Much of cancer immunotherapy is focused on engineering or activating tumor-antigen specific CD8+ T cells and, to a lesser extent, CD4+ T cells. In particular, neoantigen-specific T cells are attractive because they can kill cancer cells with high specificity. 1 A general approach starts with identifying T cells that recognize neoantigens broadly expressed within the tumor, isolating the T cells and determining their T cell receptor (TCR) sequences. These TCRs can then be transfected into patient T cells, perhaps with additional genetic engineering2 to promote more durable anti-tumor effects, and expanded into an infusion product for patient treatment. 3 In fact, this approach has recently entered the clinic, with one trial (NCT03970382) drawing from inventions from an NCI- funded CCNE U54 grant led by the PI of this proposal. 4,5 However, there are still a number of fundamental and technological challenges associated with advancing neoantigen-specific TCR-engineered therapies. First, the discovery of neoantigen-specific TCRs relies on guidance from algorithms, such as NET MHCpan 4.1, to predict antigen/MHC presentation (based upon binding and other considerations), and many neoantigens arising from truncal mutations, such as mutant KRAS or mutant TP53, are predicted as unlikely, yet have been reported as clinically effective targets. 6,7,8 Second, neoantigen-specific CD4+ T cells and their class II restricted neoantigens, while identified as important for immunotherapy-induced anti-tumor responses, 9,10 remain a largely untapped therapeutic resource, with prediction algorithms 11,12 for Class II antigen/MHC binding less developed. A third challenge is that analysis of a patient blood for neoantigen-specific T cells typically requires upwards of 20M peripheral blood mononuclear cells (PBMCs), and so isn't particularly efficient. Here we propose 3 specific Aims designed to address these outstanding issues. At the heart of the technology solutions are combinations of engineered nanoparticles (NPs) and biomolecular engineered constructs designed for efficient and selective capture, analysis, and validation of truncal neoantigen-specific CD4+ and CD8+ T cell populations. Significant preliminary data is presented for all 3 Aims, some of which uses COVID-19 patient data generated by our work during the current pandemic. 13,14 The result of this work will be a powerful toolset designed for a minimally-biased search for CD4+ and CD8+ T cell populations against truncal neoantigens (independent of patient HLA haplotype), a toolset designed for the rapid validation and characterization of those neoantigen-specific T cell clonotypes, and a public data base of Class I and Class II truncal neoantigens and T cell receptor genes specific to those neoantigens.

项目摘要 大部分癌症免疫疗法都集中在工程或激活肿瘤抗原特异性CD8+ T细胞上，以及 在较小程度上，CD4+ T细胞。特别是，新抗原特异性的T细胞很有吸引力，因为它们可以杀死 具有高特异性的癌细胞。 1一种一般方法开始识别识别新抗原的T细胞 在肿瘤内广泛表达，分离T细胞并确定其T细胞受体（TCR）序列。 然后可以将这些TCR转染到患者T细胞中，也许可以使用其他基因工程2来促进 更耐用的抗肿瘤作用，并扩展到输注产品以进行患者治疗。 3实际上，这个 最近，方法进入了诊所，一项试验（NCT03970382）从NCI-发明中绘制 由本提案的PI领导的资助CCNE U54赠款。 4,5但是，仍然有许多基本和 与推进新抗原特异性TCR工程疗法有关的技术挑战。首先， 新抗原特异性TCR的发现依赖于算法的指导，例如净MHCPAN 4.1 抗原/MHC演示（基于结合和其他考虑因素），许多新抗原是由 截短突变，例如突变的KRAS或突变体TP53，被预测为不太可能，但已报道为 临床有效目标。 6,7,8秒，新抗原特异性CD4+ T细胞及其II类限制的新抗原， 虽然认为对于免疫疗法引起的抗肿瘤反应很重要，但9,10在很大程度上尚未开发 治疗资源，具有II类抗原/MHC结合的预测算法11,12。第三 挑战是对新抗原特异性T细胞的患者血液的分析通常需要超过20m 外周血单核细胞（PBMC），因此不是特别有效。在这里，我们提出了3个特定目标 旨在解决这些杰出问题。技术解决方案的核心是 工程纳米颗粒（NP）和生物分子工程结构，旨在高效和选择性 截断新抗原特异性CD4+和CD8+ T细胞种群的捕获，分析和验证。重要的 提供了所有3个目标的初步数据，其中一些使用我们工作产生的Covid-19患者数据 在当前的大流行期间。 13,14这项工作的结果将是一个强大的工具集，专为最小偏见而设计 搜索针对截短新抗原的CD4+和CD8+ T细胞种群（与患者HLA无关 单倍型），一种旨在快速验证和表征这些新抗原特异性T细胞的工具集 clonotypes，以及I类和II类Nuncal Neoantigen和T细胞受体基因的公共数据库特异性 给那些新抗原。