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: 9981297
• 负责人: Adam M Sonabend
• 金额: $ 37.13万
• 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/9981297
• 关键词: BRAF gene; Blood - brain barrier anatomy; CAR T cell therapy; CD8-Positive T-Lymphocytes; Cell Line; Cells; Characteristics; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Development; 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; Microglia; Modeling; Molecular; Mus; Mutation; Neoplasm Transplantation; PD-1 blockade; PTPN11 gene; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Phosphorylation; Platelet-Derived Growth Factor; Process; Recurrence; Reporting; Resistance; Role; Signal Transduction; Specimen; Stains; Study models; T cell response; T-Cell Depletion; T-Lymphocyte; Therapeutic; Transgenic Mice; Transgenic Model; Transplantation; Tumor Immunity; Tumor-associated macrophages; Tumor-infiltrating immune cells; Up-Regulation; base; chemokine; chimeric antigen receptor T cells; genetic approach; glioma cell line; immunogenic; macrophage; 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.

总结 尽管在胶质母细胞瘤（GBM）中进行PD 1阻断的随机试验是阴性的，2这些患者的一个子集 确定反应的基础将导致免疫治疗有效。 用于一些患者。我们最近报告了66例复发性GBM患者的分析， PD 1阻断。1尽管根据成像标准定义缓解，但缓解患者表现出 生存率显著增加，与临床特征或额外治疗无关。30%的 应答患者的肿瘤具有BRAF和PTPN 11激活突变的富集，其刺激MAPK 因此，大多数患者对PD 1阻断反应的分子指标 仍然身份不明。我们对p38和ERK的磷酸化进行了免疫组化，这两种效应物是 这条路。这些标志物的阳性程度可预测PD 1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CD 8 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，CD 8 T细胞耗竭导致免疫原性免疫缺陷。 肿瘤在移植后，优先移植到CD 8 T细胞耗尽的受体中，而不是 免疫活性宿主。在不存在CD 8 T细胞的情况下产生的胶质瘤显示Braf上调， Ptpn 11和相关的MAPK活化，由p38和Erk的磷酸化指示。CD 8 T细胞耗竭 在神经胶质瘤发展过程中，也导致肿瘤相关巨噬细胞/小胶质细胞（TAM）的稳健增加， 促炎性TAM的基因表达。转基因小鼠MAPK活性与TAM标记的相关性 神经胶质瘤和人GBM，以及促进促炎性TAM的几种趋化因子。一个CRISPR敲- 对颅内神经胶质瘤的激酶组的筛选也涉及MAPK在肿瘤细胞的T细胞识别中的作用。这些 研究结果表明，肿瘤细胞中的MAPK活性（在神经胶质瘤进展期间被CD 8 T细胞抑制）， 促进促炎性TAM。我们推测，MAPK信号升高的GBM， CD 8 T细胞识别，因为该途径促进抗肿瘤TAM。因此，具有MAPK的GBM的子集 信号传导对PD 1阻断和CAR T细胞免疫疗法敏感。为了调查这一点，我们将SA 1） 确定胶质瘤发展过程中CD 8 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胶质瘤TAM。SA 3）确定调节胶质瘤的效果 MAPK信号传导对PD 1阻断和CAR T细胞疗法的响应。我们将使用转基因模型， 肿瘤从头发展，具有不同水平MAPK活化的细胞系，以及遗传MAPK操纵， 因为临床上可获得的药物抑制和/或促进穿过血脑屏障的MAPK信号传导。成功 这些研究的实施将确立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PD 1阻断和CAR T细胞治疗的反应。