Immunoepigenetic targeting of MHC regulators in FAP

FAP 中 MHC 调节因子的免疫表观遗传学靶向

基本信息

批准号：
10677375
负责人：
Roderick H Dashwood
金额：
$ 60.54万
依托单位：
TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677375
关键词：
Adenomatous Polyps Animals Antibodies Antigen Presentation Antineoplastic Agents ApcMin/+ mice Apoptosis Autologous Binding Biological Assay Biological Markers Biopsy CD4 Positive T Lymphocytes CD8B1 gene Cell Culture Techniques Cell Proliferation Cell Survival Cell surface Cells Cellular Structures Clinic Clinical Clinical Research Clinical Trials Coculture Techniques Colectomy Colon Colonic Neoplasms Colonic Polyps Colorectal Cancer Complex Coupled Critiques Data Diet Dose Drug Combinations Duodenal Neoplasms Duodenum Duodenum Cancer EGFR gene Ectoderm Effectiveness Embryo Epidermal Growth Factor Receptor Epigenetic Process Epithelium Erlotinib Eye Familial Adenomatous Polyposis Syndrome Female Fostering Future Gene Expression Genes Genetic Granzyme Histocompatibility Antigens Class II Histones Human Image Immune Immune Evasion Immune Targeting Immune response Immune system Immunologic Surveillance Interferons Intervention Knockout Mice Lesion Major Histocompatibility Complex Malignant Neoplasms Modeling Modification Mus Non-Steroidal Anti-Inflammatory Agents Normal Cell Operative Surgical Procedures Oral Oral Administration Organoids Patients Peptides Pharmaceutical Preparations Pharmacodynamics Pharmacotherapy Phase Polycomb Polyps Pre-Clinical Model Proteins Rattus Reader Receptor Inhibition Recommendation Regulation Research Resistance Risk Risk Reduction Safety Signal Transduction Sulindac T-Lymphocyte Testing Therapeutic Equivalency Time Tissues Titrations Toxic effect Translating Tumor Escape Up-Regulation Western Blotting Woman adaptive immune response adenoma anti-cancer cancer cell cell killing chemokine clinical care clinical translation cytokine drug candidate efficacy study epigenetic drug histone methylation human subject inhibitor innovation insight male member men mutant neoplastic next generation novel novel strategies overexpression perforin pharmacologic polyposis pre-clinical premalignant recruit response response biomarker standard of care stem transcriptomics tumor

项目摘要

Major histocompatibility complex class I and class II (MHC-I/II) members are critical for antigen presentation and adaptive immune responses but are downregulated in sporadic and genetic cases of duodenal and colorectal cancer (CRC), such as Familial Adenomatous Polyposis (FAP). There is an urgent need for novel approaches to reverse MHC-I/II silencing and overcome tumor immune evasion, starting at the adenoma stage. Based on the prior Critique, standard-of-care sulindac and erlotinib (SUL, ERL), and a new histone methyl `reader' inhibitor MAK683 that shows clinical promise, will be repurposed for immune-based interception of FAP. From the rigor of prior research and extensive preliminary data, the revised CENTRAL HYPOTHESIS is that low-dose ERL+SUL combinations along with MAK683 upregulate MHC-I/II components to enhance immune surveillance via CD8 and CD4 T cell recruitment into preneoplastic and later stages of FAP. Low-dose, non-cytotoxic test agents and their combinations augment MHC complexes at the cell surface to re-engage the host immune system for anticancer efficacy, and will be translatable to FAP patients via oral administration with a good safety margin. ✓Aim 1 MECHANISMS Aim 1A Perform dose-response and time-course studies with ERL, SUL, MAK683 and selected combinations for MHC-I/II upregulation in human and murine CRC, duodenal cancer, and adenoma cells compared with normal cells. Aim 1B Corroborate for ERL, SUL and MAK683 the increased cell surface occupancy of MHC-I/II complexes, coupled to enhanced cancer cell killing in CD8+ and CD4+ T cell co-cultures. Aim 1C Overexpress, silence, and pharmacologically inhibit mechanistic targets (e.g., EED, NLRC5, CIITA) to identify key roles in MHC-I/II regulation and immune evasion in adenoma and later stages relevant to FAP. ✓Aim 2 PRECLINICAL Aim 2A In the polyposis in rat colon (Pirc) model, perform biomarker analyses with MAK683 via i.g. drug administration and corroborate histone methylation changes and immune target modulation in colon and duodenal polyps for up to 14 days. Aim 2B Test the antitumor efficacy and safety of MAK683 alone and combined with ERL or SUL in the Pirc model. Mechanistic targets will be validated via RT-qPCR, immunoblot and multiplex CyTOF as predictors of antitumor efficacy. Aim 2C Test selected drug combinations in ApcMin/+, ApcMin/+Nlrc5–/– and ApcMin/+Ciita–/– mice for insights into Nlrc5- and Ciita-dependent/independent mechanisms. ✓Aim 3 TRANSLATIONAL Aim 3A In colon and duodenal biopsies from FAP patients, corroborate the predicted interrelationships between MHC-I/II members (B2M, CD74), coactivators (NLRC5, CIITA), mechanistic targets (EED) and epigenetic readouts (histone H3K27me3) in epithelial and immune cell components, via CyTOF. Aim 3B In FAP patient organoids, define MAK683, ERL and SUL doses that increase MHC-I/II-dependent gene expression, and examine autologous effector functions and the capacity to kill tumor organoids in T cell co- cultures. Aim 3C In organoids from Aim 3B, test the hypothesis that MAK683, ERL and SUL increase MHC-I/II- bound peptides using MS-based peptidomic approaches to aid in precision immunoepigenetic clinical strategies.

主要的组织相容性复杂I类和II类（MHC-I/II）成员对于抗原表现至关重要自适应免疫调查，但在十二指肠和结直肠癌的零星和遗传病例中被下调癌症（CRC），例如家族性腺瘤性息肉病（FAP）。迫切需要新颖的方法从腺瘤阶段开始，逆转MHC-I/II的沉默和克服肿瘤免疫进化。基于先前的评论，护理标准的苏琳克和厄洛替尼（SUL，ERL）以及一种新的组蛋白甲基“读取器”抑制剂 Mak683显示出临床承诺，将用于基于免疫的FAP拦截。从严格在先前的研究和广泛的初步数据中，修订的中心假设是低剂量ERL+SUL MAK683的组合上调上调MHC-I/II组件，以通过CD8增强免疫监视和CD4 T细胞募集到FAP的前肿瘤和后期阶段。低剂量，非毒性测试剂和它们的组合增加了细胞表面的MHC复合物，以重新接合宿主免疫系统抗癌效率，并可以通过口服给药良好的安全余量来翻译为FAP患者。 ✓IAM1机制AIM 1A对ERL，SUL，MAK683和人和鼠CRC，十二指肠癌和腺瘤中MHC-I/II上调的选定组合细胞与正常细胞相比。 AIM 1B证实了ERL，SUL和MAK683细胞表面增加 MHC-I/II复合物的占用率，与CD8+和CD4+ T细胞共培养中的癌细胞杀死相结合。 AIM 1C过表达，沉默和药物抑制机械靶标（例如EED，NLRC5，CIITA）确定在MHC-I/II/II调节中的关键作用和腺瘤中的免疫远程和与FAP相关的阶段的关键作用。 ✓aim 2大鼠结肠（PIRC）模型中多型临床前AIM 2A，使用生物标志物分析 Mak683通过I.G。药物给药和佐证组蛋白甲基化变化和免疫目标调节在结肠和十二指肠息肉中长达14天。 AIM 2B仅MAK683的抗肿瘤效率和安全性测试并与PIRC模型中的ERL或SUL结合。机械目标将通过RT-QPCR验证和多重细胞作为抗肿瘤效率的预测指标。 AIM 2C测试在APCMIN/+中选出的药物组合， apcmin/+nlrc5 - / - 以及apcmin/+ciita - / - 小鼠洞察NLRC5-和CIITA依赖/独立机制。 ✓IAM3在结肠和十二指肠活检中的翻译目标3A证实了预测的 MHC-I/II成员（B2M，CD74），共激活因子（NLRC5，CIITA）之间的相互关系，机械目标（EED）和表观遗传学读数（组蛋白H3K27ME3）在上皮和免疫细胞成分中通过Cytof。在FAP患者器官中的AIM 3B定义MAK683，ERL和SUL剂量增加了MHC-I/II依赖性基因表达，并检查自体效应子功能以及杀死T细胞中肿瘤器官的能力文化。 AIM 3B的类器官中的AIM 3C，检验Mak683，ERL和SUL增加MHC-I/II-的假设使用基于MS的Pepperidomic方法结束的胡椒体，以帮助精确免疫毛皮临床策略。