Cancer Immune-Interception for Lynch Syndrome

林奇综合征的癌症免疫拦截

• 批准号: 10706565
• 负责人: Steven M Lipkin
• 金额: $ 66.01万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-21 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706565
• 关键词: Adaptive Immune System; Affect; Affinity; Alleles; American; Antigen Presentation; Applications Grants; Aspirin; Autologous; Binding; Biological Assay; Biological Markers; CD8-Positive T-Lymphocytes; Catalogs; Cell-Mediated Cytolysis; Cells; Chemoprevention; Chemopreventive Agent; Chronic; Clinical; Clinical Trials; Clone Cells; Coculture Techniques; Colorectal; Colorectal Cancer; Coupled; Cytometry; DNA; Development; Double-Blind Method; Endometrial; Epithelial Cells; Exposure to; Familial colorectal cancer; Foundations; Frequencies; Genes; Genomics; Genotype; Germ-Line Mutation; Goals; Hereditary Malignant Neoplasm; Hereditary Nonpolyposis Colorectal Neoplasms; Image; Immune; Immune response; Immunologic Monitoring; Immunology; Immunomodulators; Immunoprevention; Immunotherapeutic agent; Incidence; Knowledge; Lesion; Life; Malignant Neoplasms; Microsatellite Repeats; Mismatch Repair; Mismatch Repair Deficiency; Mission; Mucous Membrane; Mutation; Naproxen; Non-Steroidal Anti-Inflammatory Agents; Organoids; Outcome; Ovarian; Patients; Peptide Vaccines; Peptides; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Phase Ib Clinical Trial; Population; Prevention; Prevention strategy; Prevention trial; Public Health; Randomized; Recurrence; Reporting; Research; Sampling; Screening for cancer; Small Intestines; Stains; Systems Biology; T cell receptor repertoire sequencing; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Time; Tissues; United States National Institutes of Health; Urothelium; Vaccination; Validation; adenoma; base; bioinformatics pipeline; cell killing; cell type; clinical trial implementation; co-clinical trial; cohort; colorectal cancer prevention; cytotoxicity; exome sequencing; experimental study; gene repair; genomic data; immunogenic; immunogenicity; in silico; innovation; insertion/deletion mutation; mRNA sequencing; magnetic beads; mouse model; neoantigens; neoplastic cell; novel; overexpression; peptide vaccination; premalignant; prevent; single-cell RNA sequencing; synergism; tumor; vaccination strategy

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 the DNA mismatch repair (MMR) genes. Normal colorectal epithelial cells in LS patients become MMR deficient upon acquisition of a ‘second’ somatic hit in the alternative allele of the same MMR gene that harbors the germline mutation, thus triggering the accumulation of hundreds to thousands of base-to-base mismatches and insertion-deletion mutations (indels) in microsatellite sequences. These mutations generate frameshift peptides (FSP) that become neoantigens (neoAg) and stimulate the adaptive immune system. We have reported that LS pre-cancers are immune activated and present strikingly high levels of expression of adaptive immune genes. Therefore, LS patients constitute a well-defined and prevalent population that has the potential to benefit from immune-interception strategies to prevent CRC. We have acquired a substantial amount of genomic data from LS colorectal pre-cancers and tumors to catalog and to identify the most frequent recurrent neoAg present in these lesions. In addition, we have been studying chemopreventive strategies that could augment the immune response and observed increased activation of the resident immune cells in the colorectal mucosa upon exposure to naproxen, a non-steroidal anti-inflammatory drug (NSAID), from our biomarker analysis of our NCI-sponsored Phase Ib clinical in LS patients. Furthermore, we have performed a co-clinical trial in a humanized LS mouse model that has observed that peptide vaccination with neoAg is highly effective in preventing LS CRC with the activity that is further enhanced by its combination with naproxen, thus laying the foundations for this grant proposal. The central hypothesis of this proposal is that naproxen is an immune-modulator that activates resident immune cells in the colorectal mucosa, and these will increase the recognition of NeoAg and activation of resident T-cells eliciting tumor cell killing. To explore this hypothesis, we propose three specific aims: 1. To characterize the immune cell types that are regulated after the administration of chemopreventive naproxen and aspirin in LS patients using single-cell genomics and imaging mass cytometry within a randomized phase II clinical trial; 2. To assess the immunogenicity of candidate shared neoAg identified LS patients pre-cancers and tumors for personalized immunoprevention using tetramer bound to magnetic beads in ELISpots, Tetramer stain, and cytotoxicity assays of co-cultured patient-derived organoids and autologous CD8+ T cells; 3. To profile the T cell Receptor (TCR) of neoantigen-specific CD8+ T cell clones for tracking tumor immunogenicity in LS patients. The proposed research will significantly impact the field by developing a combination of a peptide vaccination and an NSAID for immune-interception in hereditary cancers for the first time. The proposal is highly innovative by combining a chemoprevention trial using imaging mass cytometry, single-cell genomics, and systems biology to assess trial endpoints, and using tetramers bound to magnetic beads for positive selections of clones in immunology experiments.

抽象的 林奇综合征 (LS) 是遗传性结直肠癌 (CRC) 的最常见原因，影响超过 100 万人 美国人，LS 是由 DNA 错配修复 (MMR) 基因的种系突变引起的。 LS 患者的上皮细胞在获得“第二次”体细胞打击后变得 MMR 缺陷 具有种系突变的同一 MMR 基因的等位基因，从而引发数百个的积累 微卫星序列中数以千计的碱基对碱基错配和插入缺失突变 (indels)。 这些突变产生移码肽（FSP），成为新抗原（neoAg）并刺激 我们已经报道LS癌前期是激活的免疫系统并且表现得引人注目。 因此，LS 患者具有高水平的适应性免疫基因表达。 有可能受益于预防结直肠癌的免疫拦截策略的流行人群。 已从 LS 结直肠癌前期和肿瘤中获取了大量基因组数据，以进行分类和分类 以确定这些病变中最常见的复发性新抗原。此外，我们一直在研究。 化学预防策略可以增强免疫反应，并观察到免疫反应的激活增加 接触萘普生（一种非甾体抗炎药）后，结直肠粘膜中的常驻免疫细胞 药物 (NSAID)，来自我们对 NCI 赞助的 LS 患者 Ib 期临床的生物标志物分析。此外， 我们在人源化 LS 小鼠模型中进行了一项联合临床试验，观察到肽疫苗接种 与 neoAg 联合使用可非常有效地预防 LS CRC，其活性进一步增强 与萘普生，从而为这项拨款提案奠定了基础 该提案的中心假设是。 萘普生是一种免疫调节剂，可激活结直肠粘膜中的常驻免疫细胞，而这些 将增加对 NeoAg 的识别并激活驻留 T 细胞，从而引发肿瘤细胞杀伤。 根据假设，我们提出了三个具体目标： 1. 表征在 使用单细胞基因组学对 LS 患者给予化学预防性萘普生和阿司匹林 随机 II 期临床试验中的成像质谱流式细胞仪；2. 评估候选药物的免疫原性； 共享的 neoAg 鉴定了 LS 患者的癌前病变和肿瘤，使用四聚体进行个性化免疫预防 在 ELISpots、四聚体染色和共培养的患者来源的细胞毒性测定中与磁珠结合 3. 分析新抗原特异性 CD8+ T 的 T 细胞受体 (TCR)； 细胞克隆用于追踪 LS 患者的肿瘤免疫原性。这项研究将对 LS 患者的肿瘤免疫原性产生重大影响。 通过开发肽疫苗接种和 NSAID 的组合来用于遗传性免疫拦截 该提案通过结合使用成像的化学预防试验而具有高度创新性。 质谱流式分析、单细胞基因组学和系统生物学来评估试验终点，并使用四聚体结合 磁珠用于免疫学实验中克隆的阳性选择。