Cancer Immune-Interception for Lynch Syndrome

林奇综合征的癌症免疫拦截

• 批准号: 10491665
• 负责人: Steven M Lipkin
• 金额: $ 66.19万
• 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/10491665
• 关键词: Adaptive Immune System; Affect; Affinity; Alleles; American; 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; 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; Intercept; Knowledge; Lesion; Life; Malignant Neoplasms; Microsatellite Repeats; Mismatch Repair; 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; Public Health; Randomized; Recurrence; Reporting; Research; Sampling; Screening for cancer; Small Intestines; Stains; Systems Biology; T cell receptor repertoire sequencing; 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; prevent; single-cell RNA sequencing; 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）是遗传结肠癌（CRC）的最常见原因，影响> 100万 美国人。 LS是由DNA不匹配修复（MMR）基因中的种系突变引起的。正常的结直肠癌 LS患者的上皮细胞在获取替代的“第二次”体细胞后变为MMR。 具有种系突变的同一MMR基因的等位基因，从而触发了数百个的积累 在微卫星序列中，成千上万的碱到基碱不匹配和插入缺失突变（indels）。 这些突变会产生变形（FSP）的移料器，后者变成新抗原（NeoAg）并刺激 自适应免疫系统。我们报告说，LS前癌者被免疫激活，并引起了惊人的表现 适应性免疫原的高水平表达。因此，LS患者构成了一个明确的，并且 普遍的人群有可能受益于防止CRC的免疫差异策略。我们 已经从LS结直肠癌和肿瘤到目录和 确定这些病变中最常复发的NEOAG。此外，我们一直在研究 化学预防策略可以增加免疫响应并观察到增加的激活 暴露于萘普生（一种非甾体类抗炎）后，属于结直肠粘膜中的居民免疫细胞 药物（NSAID），我们对LS患者的NCI赞助的IB临床临床的生物标志物分析。此外， 我们已经在人源化的LS小鼠模型中进行了共同临床试验，该试验观察到了胡椒疫苗接种 使用NeoAg具有高效的LS CRC，其活性通过其组合进一步增强 使用萘普生，因此为该赠款提案奠定了基础。该提议的核心假设是 那萘普生是一种免疫调节剂，可激活大肠粘膜中的居民免疫细胞，而这些 将增加对NEOAG的识别和居民的激活T细胞引发肿瘤细胞的杀戮。探索这个 假设，我们提出了三个具体目标：1。表征在此之后调节的免疫细胞类型 使用单细胞基因组学和 在随机II期临床试验中成像质量细胞仪； 2。评估候选人的免疫原性 共享的NEOAG鉴定了使用四聚体的LS患者预防效果和肿瘤的肿瘤 与共同培养的患者衍生 器官和自体CD8+ T细胞； 3。介绍新抗原特异性CD8+ T的T细胞受体（TCR） 用于跟踪LS患者肿瘤免疫原性的细胞克隆。拟议的研究将极大地影响 通过开发肽疫苗的组合和用于养生的免疫障碍的NSAID 癌症是第一次。通过使用成像结合化学预防试验，该提案具有很高的创新性 质量细胞仪，单细胞基因组学和系统生物学评估试验终点，并使用四聚体结合 在免疫学实验中选择克隆正选择的磁珠。