Cancer Immune-Interception in a Spontaneous Non-Human Primate Model of Lynch Syndrome

自发性非人类灵长类动物林奇综合征模型中的癌症免疫拦截

• 批准号: 10536878
• 负责人: STANTON BRADLEY GRAY
• 金额: $ 64.63万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-22 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536878
• 关键词: Adaptive Immune System; Adjuvant; Adrenal Glands; Affect; Agonist; American; Animal Model; Antigen-Presenting Cells; Antigens; Biological Assay; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Vaccines; Catalogs; Cell Line; Cells; Chemopreventive Agent; Clinical Chemoprevention; Clinical Trials; Coculture Techniques; Code; Colorectal; Colorectal Cancer; Combined Vaccines; Coupled; Cytotoxic T-Lymphocytes; DNA; Development; Endometrium; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Familial colorectal cancer; Fostering; Genes; Genomics; Germ-Line Mutation; Goals; Hereditary Nonpolyposis Colorectal Neoplasms; Human; Immune; Immune system; Immunologic Monitoring; Immunologics; In Vitro; Incidence; Industry Collaboration; Intercept; Knowledge; Life; MHC Class I Genes; MLH1 gene; Macaca mulatta; Malignant Neoplasms; Mass Spectrum Analysis; Microsatellite Repeats; Mismatch Repair; Mission; Modeling; Mucous Membrane; Naproxen; Non-Steroidal Anti-Inflammatory Agents; Normal Cell; Outcome; Ovary; Patients; Pattern; Penetration; Peptides; Peripheral Blood Mononuclear Cell; Permeability; Phase; Physiological; Play; Population; Population Genetics; Prevention; Preventive vaccine; Public Health; Publishing; Recurrence; Reporting; Research; Rhesus; Safety; Small Intestines; Stomach; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Tissues; Toll-like receptors; Translations; United States National Institutes of Health; Vaccination; Vaccines; Validation; base; bioinformatics pipeline; cancer cell; cancer prevention; cancer type; cell killing; co-clinical trial; cohort; cytokine; cytotoxic; cytotoxic CD8 T cells; cytotoxicity; first-in-human; gene repair; high risk; immune activation; immune stimulant; immunogenic; immunogenicity; innovation; insertion/deletion mutation; neoantigens; next generation sequencing; nonhuman primate; novel; response; tool; translation to humans; tumor; vaccine development; vaccine platform

ABSTRACT Lynch Syndrome (LS) is the most common cause of hereditary colorectal cancer (CRC) affecting >1 million Americans. LS is caused by germline mutations in one of four DNA mismatch repair (MMR) genes. Normal colorectal epithelial cells in LS patients become MMR deficient after a somatic ‘second’ hit generating the accu- mulation of hundreds of insertion-deletion mutations (indels) in microsatellite sequences. These indels generate frameshift peptides (FSP) that become neoantigens (neoAg) and stimulate the adaptive immune system. We have previously reported that adaptive immune genes are highly expressed in LS pre-cancers, and we have generated a detailed neoAg catalog with >1,000 FSP neoAg from a cohort of LS pre-cancers and early-stage CRCs using next-generation sequencing tools coupled with a state-of-the-art bioinformatics pipeline. In addition, we have published the results of a phase Ib chemoprevention clinical trial in LS patients using naproxen showing immune-activation of colorectal mucosa resident cells. Taken together, these results point strongly towards the development of a vaccine for ‘recurrent’ and ‘shared’ LS-associated tumor neoAg combined with naproxen for pan-cancer prevention in the LS population. However, the main knowledge gap remains to select the most opti- mal neoAg peptides and to establish the efficacy and safety of the vaccination in a reliable animal model that allows immediate human translation. Rhesus macaques are a promising non-human primate (NHP) model dis- playing the closest genomic resemblance to humans. Our research team has reported the first colony of spon- taneous LS in rhesus carrying a germline mutation in MLH1. In addition, we are partnering with industry collab- orators in AMAL Therapeutics that have developed an innovative vaccination platform called KISIMA, integrating several selected FSP in tandem with a cell-permeable peptide fostering cell penetration, and a toll-like receptor agonist that acts as a self-adjuvant. Our central hypothesis is that our state-of-the-art bioinformatics pipeline for neoAg prediction will lead to the identification of the most immunogenic, recurrent across tumors, and shared among LS-associated tumor types FSP neoAg to be integrated in the KISIMA self-adjuvant vaccine platform, which will render a strong immunogenicity in combination with naproxen. To explore this hypothesis, we propose three specific aims: 1. To validate in vitro the immunogenicity of the top 150 recurrent neoAg shared by LS non-colorectal tumors using ELISpot, ELISA, and cytokine assays using PBMCs and CD8+ T cells from healthy human donors; 2. To develop artificial antigen-presenting cells (aAPC) expressing human LS neoAg to validate the cytotoxicity of neoAg-enriched T cells; and 3. To assess the immunogenicity of a neoAg combination using the novel self-adjuvant vaccine platform KISIMA alone and in combination with naproxen in a co-clinical trial in LS rhesus. The proposal is highly innovative because is developing a novel self-adjuavnt vaccine platform in a unique spontaneous NHP model of LS. The proposed research will significantly impact the field because it is a stepping stone to develop a Phase I first-in-human clinical trial to test a novel CRC vaccine for LS patients.

抽象的 林奇综合征（LS）是遗传结肠癌（CRC）的最常见原因，影响> 100万 美国人。 LS是由四个DNA不匹配修复（MMR）基因之一中的种系突变引起的。普通的 LS患者的结直肠上皮细胞在体细胞“第二次命中”后变为MMR缺乏 微卫星序列中数百个插入 - 缺失突变（indels）的插入。这些indels会产生 移植肽（FSP）变成新抗原（NEOAG）并刺激适应性免疫学系统。我们 以前曾报道过，自适应免疫基因在LS前癌中高度表达，我们有 从LS前癌症和早期阶段产生了一个详细的NEOAG目录，其中> 1,000 FSP NEOAG。 CRC使用下一代测序工具，再加上最先进的生物信息学管道。此外， 我们已经使用萘普生显示了LS患者的IB期化学预防临床试验的结果 结直肠粘膜居民细胞的免疫激活。综上所述，这些结果强烈指向 开发用于“复发”和“共享” LS相关肿瘤NEOAG与萘普生的疫苗的开发 LS人群中的泛伴侣预防。但是，主要知识差距仍然是选择最选择的 mal neog petides并在可靠的动物模型中建立疫苗接种的效率和安全性 允许直接的人类翻译。恒河猕猴是一种有希望的非人类灵长类动物（NHP）模型 与人类最接近基因组相似。我们的研究团队报告了赞助商的第一个殖民地 - 在MLH1中携带种系突变的恒河中的ls。此外，我们正在与行业合作合作 - Amal Therapeutics的演说家开发了一种创新的疫苗平台，称为Kisima，整合 与细胞渗透的细胞培养细胞渗透的串联中有几个选定的FSP，并且类似收费的接收器 充当自动的激动剂。我们的中心假设是我们最先进的生物信息学管道 对于NEOAG预测，将导致识别最免疫原性，跨肿瘤的复发，并共享 在与LS相关的肿瘤类型中，FSP NEOAG集成在Kisima自动疫苗平台中， 这将与萘普生结合使用强大的免疫原性。为了探讨这一假设，我们提出了 三个具体的目的：1。验证在体外LS共享的前150个复发性NEOAG的免疫原性 使用ELISPOT，ELISA和细胞因子测定的非色直肠肿瘤，使用PBMC和CD8+ T细胞来自健康 人类捐助者； 2。开发表达人LS NEOAG的人工抗原细胞（AAPC）以验证 富含NEOAg的T细胞的细胞毒性；和3。使用使用NEOAG组合的免疫原性 新型的自我辅助疫苗平台Kisima单独并与Naproxen结合在一项共同临床试验中 LS恒河。该提案具有很高的创新性 LS的独特赞助NHP模型。拟议的研究将显着影响该领域，因为它是 垫脚石开发了I期第一阶段的人类临床试验，以测试LS患者的新型CRC疫苗。