Immunoepigenetic targeting of MHC regulators in FAP

FAP 中 MHC 调节因子的免疫表观遗传学靶向

• 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
• 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

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.

主要组织相容性复合体I类和II类（MHC-I/II）成员对于抗原呈递和免疫应答是关键的。 获得性免疫应答，但在散发性和遗传性十二指肠和结直肠癌病例中下调 癌症（CRC），如家族性腺瘤性息肉病（FAP）。迫切需要新的方法 逆转MHC-I/II沉默和克服肿瘤免疫逃避，从腺瘤阶段开始。基于 先前的批评，标准护理舒林酸和厄洛替尼（SUL，ERL），和一种新的组蛋白甲基“阅读器”抑制剂 显示出临床前景的MAK 683将被重新用于基于免疫的FAP拦截。从尸体僵硬的程度来看 根据先前的研究和广泛的初步数据，修订后的中心假设是，低剂量ERL+SUL 与MAK 683的组合沿着上调MHC-I/II组分以增强通过CD 8 和CD 4 T细胞募集到FAP的癌前和晚期。低剂量、无细胞毒性试验药物和 它们的组合增加了细胞表面的MHC复合物，以重新参与宿主免疫系统， 因此，本发明的组合物具有抗癌功效，并且将可经由口服施用以良好的安全裕度转移至FAP患者。 目的1A用ERL、SUL、MAK 683和MAK 684进行剂量反应和时间过程研究。 用于人和鼠CRC、十二指肠癌和腺瘤中MHC-I/II上调的选定组合 与正常细胞相比。目的1B Coroborate对ERL、SUL和MAK 683细胞表面的增加 MHC-I/II复合物的占据，与CD 8+和CD 4 + T细胞共培养物中增强的癌细胞杀伤偶联。 目的1C过表达、沉默和抑制机制靶点（例如，EED、NLRC 5、CIITA）至 确定MHC-I/II调节和免疫逃避在腺瘤中的关键作用，以及与FAP相关的后期阶段。 目的2临床前目的2A在大鼠结肠息肉病（Pirc）模型中，使用以下进行生物标志物分析： MAK 683通过i.g.药物给药和证实组蛋白甲基化变化和免疫靶点调节 在结肠和十二指肠息肉中长达14天。目的2B检测MAK 683单药抗肿瘤的有效性和安全性 并在Pirc模型中与ERL或SUL相结合。机制靶标将通过RT-qPCR、免疫印迹进行验证 和多重CyTOF作为抗肿瘤功效的预测因子。目标2C在ApcMin/+中测试选定的药物组合， ApcMin/+ Nlrc 5-/-和ApcMin/+Ciita-/-小鼠，以深入了解Nlrc 5-和Ciita-依赖性/非依赖性机制。 目标3目标3A在FAP患者的结肠和十二指肠活检中，证实了预测的 MHC-I/II成员（B2 M、CD 74）、辅激活因子（NLRC 5、CIITA）、机制靶点之间的相互关系 (EED)和上皮和免疫细胞组分中的表观遗传读数（组蛋白H3 K27 me 3）。 目的3B在FAP患者类器官中，确定增加MHC-I/II依赖性基因表达的MAK 683、ERL和SUL剂量。 表达，并检查自体效应子功能和T细胞共刺激中杀死肿瘤类器官的能力。 cultures.在来自目标3B的类器官中，检验MAK 683、ERL和SUL增加MHC-I/II的假设。 使用基于MS的肽组学方法结合肽，以帮助精确的免疫表观遗传临床策略。