Small-molecule Inhibition of Glycogen Synthase Kinase-3 Sensitizes Colorectal Cancer Cells and Stimulates Immune Cells to Bolster Immune Cell-mediated Tumor Cytotoxicity

糖原合酶激酶 3 的小分子抑制使结直肠癌细胞敏感并刺激免疫细胞增强免疫细胞介导的肿瘤细胞毒性

• 批准号: 10594422
• 负责人: Kelsey Elizabeth Huntington
• 金额: $ 2.39万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10594422
• 关键词: Address; Biological Assay; Biological Markers; CT26; Cancer Biology; Cancer Center; Cancer Model; Cell-Mediated Cytolysis; Cells; Cessation of life; Coculture Techniques; Colon Carcinoma; Colorectal Cancer; Combination immunotherapy; Combined Modality Therapy; Communication; Disease; Educational process of instructing; Environment; Evaluation; Experimental Designs; Fellowship; Flow Cytometry; Fluorescence Microscopy; Genomics; Glycogen Synthase Kinase 3; Goals; Group Meetings; Immune; Immunocompetent; Immunohistochemistry; Immunologic Cytotoxicity; Immunologic Monitoring; Immunologic Stimulation; In Vitro; Inbred BALB C Mice; Incidence; Inflammation Mediators; Invaded; Journals; Knowledge; Ligands; Literature; Mediating; Mentorship; Microsatellite Instability; Microsatellite Repeats; Microscopy; Monitor; Mouse Strains; Mus; Patients; Property; Receptor Signaling; Research; Research Design; Role; Signal Pathway; Signal Transduction; Solid Neoplasm; Technical Expertise; Therapeutic; Therapeutic Effect; Toxic effect; Training; Translational Research; Tumor Burden; Tumor Cell Line; Tumor Immunity; Universities; Western Blotting; Work; anti-PD-1; anti-PD-L1; anti-tumor immune response; bioimaging; cancer cell; career development; cell killing; checkpoint receptors; clinical translation; comparison control; cytokine; cytotoxicity; experimental study; immune checkpoint; immune checkpoint blockade; immunoregulation; improved; in vivo; inhibitor; insight; meetings; mortality; mouse model; neoplastic cell; novel; programs; receptor expression; response; skills; small molecule; synergism; transcriptome sequencing; tumor; tumor immunology

PROJECT SUMMARY Globally, colorectal cancer (CRC) ranks third in incidence and second in mortality. Immune checkpoint blockade (ICB) has demonstrated impressive efficacy in microsatellite instability (MSI)-positive CRC, however, there remains a substantial unmet need for 96% of patients with microsatellite stable (MSS) advanced CRC who don’t respond. A growing literature supports an immunomodulatory role of glycogen synthase kinase-3 (GSK-3) in the context of anti-tumor immunity. The proposed research focuses on GSK-3 inhibitor 9-ING-41. We hypothesize that GSK-3 inhibition can improve efficacy of ICB by (1) activating immune cells via increased inflammatory mediators and decreased checkpoint receptor expression and by (2) sensitizing tumor cells to immune cell-mediated tumor cell killing via increased death receptor expression and signaling, increased checkpoint ligand expression, and decreased survival pathway signaling. In year one, Aim 1 experiments will examine the potential of 9-ING-41 to promote anti-tumor immunity, modify NF-κB signaling, and regulate immune checkpoint expression in vitro. In our preliminary evaluation of 9-ING-41 in a co-culture assay we observed increased immune cell-mediated tumor cell killing with 9-ING-41, compared to control. We will analyze cytokine profiles of tumor and immune cells treated with 9-ING-41 to make predictions about therapeutic impact on anti-tumor immunity. We plan to evaluate checkpoint ligand expression and downstream targets of NF-κB signaling using western blots, RNA-seq, and flow cytometry in 9-ING-41-treated cells. In year two, Aim 2 experiments will utilize an immunocompetent, syngeneic murine CRC model to monitor immune cell invasion and activation, and tumor cell killing in response to 9-ING-41 monotherapy or combination ICB therapy with αPD- 1 or αPD-L1. This work will be carried out in Dr. Wafik El-Deiry’s lab, which has expertise in CRC, cytokine profiling, in vivo experimental design, biomarkers, and clinical translation. Because the El-Deiry Lab is located at Brown University, we have access to state-of-the-art bioimaging, genomics, microscopy, and flow cytometry facilities. The Cancer Center at Brown, located within the same building as the lab, offers the applicant access to invited speaker seminars, research program meetings, and translational research disease group meetings, while the Pathobiology Program offers invited speaker seminars and journal clubs. This environment enables the training plan by facilitating the applicant’s new technical skills, cancer biology and immunology expertise, and through individualized mentorship, teaching, and scientific communication opportunities. The project will lead to the applicant’s mastery of new technical and research design skills in addition to career development included within the fellowship training plan. This novel research will offer mechanistic insights into the immunomodulatory mechanisms of GSK-3 inhibitors and will address the significant unmet need of effective immunotherapy combinations for the vast majority of patients with CRC who don’t respond to ICB therapy.

项目摘要 在全球范围内，大肠癌（CRC）在发病率中排名第三，死亡率排名第二。免疫检查点封锁 （ICB）证明了微卫星不稳定性（MSI） - 阳性CRC的效率令人印象深刻 对于96％的微卫星稳定患者（MSS）晚期CRC，仍然没有满足的需求 回应。越来越多的文献支持糖原合酶激酶3（GSK-3）在 抗肿瘤免疫学的背景。拟议的研究重点是GSK-3抑制剂9-ING-41。我们假设 GSK-3抑制可以通过（1）通过增加激活免疫球来提高ICB的效率 炎症介质和改善的检查点受体表达以及（2）敏化肿瘤 通过增加的死亡受体表达和信号传导来进行免疫细胞介导的肿瘤细胞杀死的细胞， 增加了检查点配体表达，并增加了存活途径信号传导。在第一年，目标1 实验将检查9-ING-41的潜力，以促进抗肿瘤免疫史，修改NF-κB信号传导和 在体外调节免疫检查点表达。在我们对共培养测定中对9-ing-41的初步评估中 与对照相比，我们观察到使用9-ING-41的免疫细胞介导的肿瘤细胞杀死增加。我们将 分析用9-ING-41处理的肿瘤和免疫细胞的细胞因子谱，以预测治疗 对抗肿瘤免疫的影响。我们计划评估检查点配体的表达和下游目标 使用蛋白质印迹，RNA-seq和流式细胞仪中的NF-κB信号传导，在9-ENG-41处理的细胞中。在第二年，目标2 实验将利用免疫能力的，合成的鼠CRC模型来监测免疫球体入侵 和激活，以及响应9-ING-41单一疗法或ICB治疗的肿瘤细胞杀死 1或αPD-L1。这项工作将在Wafik El-Deiry博士的实验室中进行，该实验室在CRC中具有专业知识，Cytokine 分析，体内实验设计，生物标志物和临床翻译。因为El-Deiry实验室位于 在布朗大学，我们可以使用最先进的生物成像，基因组学，显微镜和流式细胞仪 设施。布朗的癌症中心与实验室在同一建筑物内，提供适用的访问 邀请演讲者的半手，研究计划会议和翻译研究疾病小组会议， 病理生物学计划提供受邀演讲者和期刊俱乐部。这个环境可以启用 培训计划通过促进申请人的新技术技能，癌症生物学和免疫学专业知识， 并通过个性化的训练，教学和科学沟通机会。该项目将 除了职业发展外，还导致申请人掌握新的技术和研究设计技能 包括在奖学金培训计划中。这项新颖的研究将提供有关机械的见解 GSK-3抑制剂的免疫调节机制，将满足有效的重要需求 绝大多数不对ICB治疗反应的CRC患者的免疫疗法组合。