PIKfyve regulation of IL-12 signaling in dendritic cells and cancer immunity

PIKfyve 对树突状细胞中 IL-12 信号传导和癌症免疫的调节

基本信息

批准号：
10451543
负责人：
Jae Eun Choi
金额：
$ 2.02万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10451543
关键词：
Ablation Affect Antigens Antiviral Agents Autoimmune Automobile Driving Biological Biological Response Modifiers Biology Bone Marrow CD8-Positive T-Lymphocytes Cancer Model Cancer Patient Cells Cessation of life Clinic Clinical Clinical Trials Communicable Diseases Complex Data Dendritic Cells Disease Endosomes Enzyme-Linked Immunosorbent Assay Feedback Flow Cytometry Gene Proteins Genetic Genetic Transcription Goals IL12B gene Immune checkpoint inhibitor Immune system Immunotherapy In Vitro Interleukin-10 Interleukin-12 Knock-out Knowledge Link Lipid III Lipids MC38 Malignant Neoplasms Measures Mediating Methods Modeling Monitor Mus Nature Oral Pathway interactions Patients Pharmaceutical Preparations Pharmacology Phase II Clinical Trials Phosphatidylinositols Phosphotransferases Protein Subunits Proteins Regulation Research Proposals Role Signal Pathway Signal Transduction Signaling Molecule T cell response T-Cell Activation T-Lymphocyte Transcriptional Regulation Translating Tumor Immunity Vaccines Vacuole Viral Wild Type Mouse Work adaptive immunity anti-PD-1 anti-PD1 therapy anti-cancer anti-tumor immune response antitumor effect cancer clinical trial cancer immunotherapy cancer therapy conditional knockout cytokine cytotoxic draining lymph node drug efficacy effector T cell enzyme linked immunospot assay experimental study improved in vivo inhibition of autophagy inhibitor inorganic phosphate insight kinase inhibitor knock-down late endosome mouse model neoantigens novel pathogen phosphatidylinositol 3-phosphate pre-clinical preclinical study response success targeted treatment therapeutic target transcriptome sequencing tumor tumor growth tumor immunology tumor microenvironment tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT Since the discovery of the IL-12 cytokine, it has been considered a “master regulator” of the immune system. Primarily produced by dendritic cells, IL-12 is the key signaling molecule linking innate to adaptive immunity to drive IFN𝛾𝛾/Th1-mediated T cell responses against pathogens and cancer. Preclinical studies have long shown that enhancing IL-12 signaling improves vaccines. Recently, IL-12 in dendritic cells was shown to be required for anti-PD1 efficacy in mouse models of cancer. Despite the data nominating the IL-12 pathway as a therapeutic target in cancer, previous attempts to directly administer IL-12 in clinical trials have resulted in patient death from cytotoxic response or poor efficacy due to IL-10-mediated negative feedback. Subsequent efforts in cancer have largely focused on improving methods to deliver and release IL-12 with limited success. Therefore, there is a significant need to better understand the biology of IL-12 regulation in dendritic cells to identify novel, safe and effective strategies of targeting this pathway as an anti-cancer strategy. Our group previously identified PIKfyve lipid kinase as a novel target of the highly efficacious, well- tolerated and orally administered multi-kinase inhibitor, ESK981. We demonstrated the ability of this drug to reduce tumor growth in multiple syngeneic mouse models of cancer and enhance anti-PD1 therapy. Interestingly, drug efficacy was dependent on the presence of CD8+ T cells, and the IL-12 and IFN𝛾𝛾 and signaling pathways in vivo. Furthermore, ESK981 could directly enhance IL-12 signaling in primary dendritic cells in vitro and in vivo through transcriptional regulation of IL12B. Therefore, the overall goal of this project is to define the mechanism of IL-12 regulation by PIKfyve and demonstrate the nature of PIKfyve inhibition, as an anti-cancer therapy, in anti-tumor immune responses. The overarching hypothesis is that PIKfyve inhibition induces IL-12 signaling in dendritic cells to enhance antigen- specific, CD8+ T cell responses in cancer. Experiments in Aim 1 will investigate the effect of Pikfyve knockdown and conditional knock-out in mouse dendritic cells on the genes and protein subunits in the IL-12 signaling pathway in vitro and in vivo. To examine dendritic cell subsets, such as CD8α+ and CD103+ cDCs, mice with genetic knock-out of Batf3 will be included for in vitro and in vivo studies. Experiments in Aim 2 will study the potential anti-tumor effect of PIKfyve inhibitors, including ESK981and Apilimod, on dendritic cell Pikfyve conditional knock-out, Batf3 knock-out, and wild type mice. We will monitor tumor progression and examine T cell responses in the tumor draining lymph nodes and tumor microenvironment, including OVA model antigen and neoantigen specific CD8+ T cell activation using tetramers available for the MC38 tumor model. The proposed study will provide greater insight into the biology of IL-12 signaling in dendritic cells and could provide the rationale for selecting PIKfyve inhibitors as safe, effective methods of enhancing IL-12 alone and in combination with IL-12-dependent therapies, such as checkpoint inhibitors and vaccines in cancer.

项目摘要/摘要自从发现IL-12细胞因子以来，它被认为是免疫的“主调节器” 系统。 IL-12主要由树突状细胞产生，是将先天物与自适应联系起来的关键信号分子驱动IFN𝛾𝛾/Th1介导的T细胞反应的免疫力针对病原体和癌症。临床前研究已有长期表明，增强IL-12信号传导可改善疫苗。最近，树突状细胞中的IL-12显示为在癌症小鼠模型中抗PD1效率所必需。尽管数据提名了IL-12途径作为癌症的治疗靶标，先前在临床试验中直接管理IL-12的尝试导致因IL-10介导的负反馈而导致的细胞毒性反应或效率差的患者死亡。随后的癌症的努力主要集中在改善成功交付和释放IL-12的方法上，成功有限。因此，非常需要更好地了解树突状细胞中IL-12调节的生物学确定针对这种途径的新颖，安全有效的策略，作为一种反癌症策略。我们的小组先前将pikfyve脂质激酶确定为高效，良好的新目标耐受和口服的多激酶抑制剂ESK981。我们证明了这种药物的能力在多种癌症的多种合成小鼠模型中降低肿瘤的生长并增强抗PD1治疗。有趣的是，药物效率取决于CD8+ T细胞的存在，IL-12和IFN𝛾𝛾和体内信号通路。此外，ESK981可以直接增强一级树突中的IL-12信号传导通过IL12B的转录调节体外和体内细胞。因此，该项目的总体目标是定义Pikfyve和Pikfyve和在抗肿瘤免疫反应中证明了pikfyve抑制作用，作为一种抗癌治疗。总体假设是pikfyve抑制作用诱导树突状细胞中的IL-12信号传导，以增强抗原癌症中的特异性CD8+ T细胞反应。 AIM 1中的实验将研究Pikfyve的效果 IL-12中的基因和蛋白质亚基的小鼠树突状细胞中的敲低和条件敲除体外和体内信号通路。要检查树突状细胞亚群，例如CD8α+和CD103+ CDC，具有BATF3遗传敲除的小鼠将包括用于体外和体内研究。 AIM 2中的实验将研究pikfyve抑制剂（包括ESK981和apilimod）对树突状细胞的潜在抗肿瘤作用 Pikfyve有条件的淘汰赛，BATF3淘汰赛和野生型小鼠。我们将监测肿瘤的进展和检查肿瘤排出淋巴结和肿瘤微环境中的T细胞反应，包括OVA 使用可用于MC38肿瘤的四聚体的抗原和新抗原特异性CD8+ T细胞激活模型。拟议的研究将为树突状细胞和可以提供选择Pikfyve抑制剂作为安全，有效的方法的基本原理并结合IL-12依赖性疗法，例如检查点抑制剂和癌症中的疫苗。