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

• 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
• 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

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 尽管胶质母细胞瘤 (GBM) 中 PD1 阻断的随机试验结果为阴性，2 其中一部分患者 对免疫治疗有反应。3 确定反应的基础将使免疫治疗有效 用于某些患者。我们最近报告了对 66 名接受过治疗的复发性 GBM 患者的分析 PD1 阻断。1 虽然反应是根据影像学标准定义的，但有反应的患者表现出 生存率显着提高，且与临床特征或额外治疗无关。 30%的 反应性患者的肿瘤富含 BRAF 和 PTPN11 激活突变，可刺激 MAPK 通路信号传导。1 因此，大多数患者对 PD1 阻断反应的分子指标 仍然身份不明。我们对 p38 和 ERK（两个效应子）的磷酸化进行了免疫组织化学分析。 这条途径。这些标志物的阳性程度可预测 PD1 阻断后的总生存期。的 与我们的患者肿瘤研究相关，我们模拟了 CD8 T 细胞耗竭对肿瘤进展的影响 从头发育的小鼠转基因神经胶质瘤。在这种情况下，CD8 T 细胞耗竭导致免疫原性 移植后，优先植入 CD8 T 细胞耗尽的受体中的肿瘤，而不是 免疫能力强的宿主。在缺乏 CD8 T 细胞的情况下产生的胶质瘤显示 Braf 和 Ptpn11 以及 p38 和 Erk 磷酸化表明的相关 MAPK 激活。 CD8 T 细胞耗竭 在神经胶质瘤发展过程中，还导致肿瘤相关巨噬细胞/小胶质细胞（TAM）的强劲增加，并且 促炎 TAM 的基因表达。转基因小鼠中 MAPK 活性与 TAM 标记相关 神经胶质瘤和人类 GBM，并具有多种促进促炎 TAM 的趋化因子。 CRISPR敲除—— 颅内神经胶质瘤激酶组的筛选也表明 MAPK 参与了 T 细胞对肿瘤细胞的识别。这些 研究结果表明肿瘤细胞中的 MAPK 活性（在神经胶质瘤进展过程中被 CD8 T 细胞抑制）， 促进促炎 TAM。我们假设 MAPK 信号传导升高的 GBM 更容易受到影响 CD8 T 细胞识别，因为该途径可促进抗肿瘤 TAM。因此，具有 MAPK 的 GBM 子集 信号传导对 PD1 阻断和 CAR T 细胞免疫疗法敏感。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 MAPK 信号传导对 PD1 阻断和 CAR T 细胞治疗的反应。我们将使用转基因模型，其中 肿瘤从头发育，细胞系具有不同水平的 MAPK 激活，以及遗传 MAPK 操作 因为临床上可用的药物会抑制和/或促进穿过血脑屏障的 MAPK 信号传导。成功的 这些研究的执行将确定 MAPK 信号在神经胶质瘤中的新的免疫相关作用。我们将 确定肿瘤细胞的 MAPK 信号传导是否影响对 PD1 阻断和 CAR T 细胞治疗的反应。