MAPK as target of glioma immunoediting by CD8 T-cells, and predictor of response to immunotherapy

MAPK 作为 CD8 T 细胞神经胶质瘤免疫编辑的靶点以及免疫治疗反应的预测因子

基本信息

批准号：
10542819
负责人：
Adam M Sonabend
金额：
$ 37.6万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-15 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10542819
关键词：
BRAF gene CAR T cell therapy CD8-Positive T-Lymphocytes Cd68 Cell Line Cells Characteristics Clinical Clustered Regularly Interspaced Short Palindromic Repeats Development Engraftment Exhibits Flow Cytometry Gene Expression Genetic Glioblastoma Glioma Human ITGAM gene Image Immune Immunocompetent Immunohistochemistry Immunotherapy Implant Infiltration Inflammatory Knock-out Knockout Mice MAP Kinase Gene MAPK Signaling Pathway Pathway Macrophage Microglia Modeling Molecular Mus Mutation Neoplasm Transplantation PD-1 blockade PTPN11 gene Pathway interactions Patients Pharmaceutical Preparations Phenotype Phosphorylation Platelet-Derived Growth Factor Predisposition Process Recurrence Reporting Resistance Role Signal Transduction Specimen Stains T cell response T-Cell Depletion T-Lymphocyte Therapeutic Transgenic Mice Transgenic Model Transplantation Tumor Immunity Tumor-associated macrophages Up-Regulation blood-brain barrier crossing cancer infiltrating T cells chemokine chimeric antigen receptor T cells genetic approach glioma cell line immunogenic neoplastic cell novel p38 Mitogen Activated Protein Kinase patient response patient subsets predicting response programmed cell death protein 1 randomized trial response tumor tumor progression

项目摘要

SUMMARY Whereas randomized trials for PD1 blockade in glioblastoma (GBM) are negative,2 a subset of these patients do respond to immunotherapy.3 Determining the basis of response will result in immunotherapy being effectively used for some patients. We recently reported the analysis of 66 recurrent GBM patients that were treated with PD1 blockade.1 Whereas response was defined based on imaging criteria, responsive patients exhibited significant increased survival that was independent of clinical characteristics or additional treatments. 30% of tumors of responsive patients had enrichment of BRAF and PTPN11 activating mutations, which stimulate MAPK pathway signaling.1 Therefore, molecular indicators of response to PD1 blockade for the majority of patients remain unidentified. We performed immunohistochemistry for phosphorylation of p38 and ERK, two effectors of this pathway. Extent of positivity for these markers was predictive of overall survival following PD1 blockade. Of relevance to our patient tumor studies, we modeled the effect of CD8 T-cell depletion on tumor progression on mouse transgenic gliomas that develop de novo. In this setting, CD8 T-cell depletion resulted in immunogenic tumors that upon transplantation, preferentially engrafted in recipients with CD8 T-cell depletion as opposed to immunocompetent hosts. Gliomas generated in the absence of CD8 T-cells showed upregulation of Braf and Ptpn11, and associated MAPK activation indicated by phosphorylation of p38 and Erk. CD8 T-cell depletion during glioma development also led to robust increase in tumor associated macrophages/microglia (TAM), and gene expression of pro-inflammatory TAM. MAPK activity correlated with TAM markers in mouse transgenic gliomas and human GBM, and with several chemokines that promote pro-inflammatory TAM. A CRISPR knock- out screen for the kinome on intracranial gliomas also implicated MAPK in T-cell recognition of tumor cells. These findings suggest that MAPK activity in tumor cells (that is suppressed by CD8 T-cells during glioma progression), promotes pro-inflammatory TAM. We hypothesize that GBM with elevated MAPK signaling, are more susceptible to CD8 T-cell recognition, as this pathway promotes anti-tumoral TAM. Thus, the subset of GBM that have MAPK signaling are susceptible to PD1 blockade and CAR T-cell immunotherapy. To investigate this, we will SA1) Determine the effects of CD8 T-cell depletion during glioma development on TAM phenotype. SA 2) Determine whether MAPK signaling promotes anti-tumoral glioma TAM. SA 3) Determine the effect of modulating glioma MAPK signaling on response to PD1 blockade and CAR T-cell therapy. We will use transgenic models in which tumors develop de novo, cell lines with varying levels of MAPK activation, genetic MAPK manipulation as well as clinically available drugs inhibit and/or promote MAPK signaling that cross the blood-brain barrier. Successful execution of these studies will establish a novel immune-related role of MAPK signaling in glioma. We will determine whether MAPK signaling by tumor cells influences response to PD1 blockade, and CAR T-cell therapy.

概括而胶质母细胞瘤（GBM）中PD1阻断的随机试验为阴性，2这些患者的一部分对免疫疗法做出反应。3确定反应的基础将有效地导致免疫疗法用于某些患者。我们最近报道了对接受治疗的66例复发性GBM患者的分析 PD1封锁。1基于成像标准定义了反应，而反应敏感的患者表现出与临床特征或其他治疗无关的生存率显着增加。 30％反应敏感患者的肿瘤具有BRAF和PTPN11激活突变的富集，这会刺激MAPK 1因此，大多数患者对PD1阻断的反应的分子指标保持身份不明。我们对p38和ERK的磷酸化进行了免疫组织化学，两个这条路。这些标记的阳性程度可预测PD1阻滞后的总生存率。的与我们的患者肿瘤研究相关，我们建模了CD8 T细胞耗竭对肿瘤进展的影响小鼠转基因神经胶质瘤，从头发展。在这种情况下，CD8 T细胞耗竭导致免疫原性移植后的肿瘤，优先植入CD8 T细胞耗竭的受体中免疫能力的宿主。在没有CD8 T细胞的情况下产生的神经膜瘤显示BRAF和 PTPN11以及相关的MAPK激活，用p38和ERK的磷酸化表示。 CD8 T细胞耗竭在神经胶质瘤发育过程中，还导致肿瘤相关巨噬细胞/小胶质细胞（TAM）和促炎TAM的基因表达。 MAPK活动与小鼠转基因中的TAM标记相关神经胶质瘤和人类GBM，并带有几种促进促炎性TAM的趋化因子。 crispr敲门颅内神经胶质瘤上的Kinome的外屏还暗示了MAPK在肿瘤细胞的T细胞识别中。这些研究结果表明，MAPK活性在肿瘤细胞中（在神经胶质瘤进展过程中被CD8 T细胞抑制），促进促炎性TAM。我们假设具有升高MAPK信号的GBM更易感随着该途径促进抗肿瘤TAM，CD8 T细胞识别。因此，具有MAPK的GBM的子集信号传导容易受到PD1封锁和CAR T细胞免疫疗法的影响。为了调查这一点，我们将SA1）确定神经胶质瘤发育过程中CD8 T细胞耗竭对TAM表型的影响。 SA 2）确定 MAPK信号传导是否促进抗肿瘤神经胶质瘤TAM。 SA 3）确定调节神经胶质瘤的效果对PD1封锁和CAR T细胞治疗的反应的MAPK信号传导。我们将使用转基因模型肿瘤发展起来，MAPK激活水平不同的细胞系，基因MAPK操纵由于临床上可用的药物抑制和/或促进跨越血脑屏障的MAPK信号传导。成功的这些研究的执行将建立MAPK信号在神经胶质瘤中的新型免疫相关作用。我们将确定肿瘤细胞的MAPK信号是否影响对PD1阻滞的反应和CAR T细胞治疗。