Bone marrow-derived myeloid cell dysregulation in malignant progression of glioma

胶质瘤恶性进展中骨髓源性骨髓细胞失调

• 批准号: 10730970
• 负责人: Prajwal Rajappa
• 金额: $ 47.03万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730970
• 关键词: Adolescent and Young Adult; Affect; Automobile Driving; Blood; Bone Marrow; Bone Marrow Cells; CD8B1 gene; Cell Differentiation process; Cell Separation; Cells; Central Nervous System Neoplasms; Circulation; Cytometry; Data; Dependence; Flow Cytometry; Genes; Glioma; Goals; Human; ID2 gene; Immunologic Surveillance; Immunosuppression; Impairment; Infiltration; Knock-out; Macrophage; Malignant - descriptor; Malignant Glioma; Methods; Migration Inhibitory Factor; Modeling; Mus; Myelogenous; Myeloid Cells; Myeloid-derived suppressor cells; Natural Killer Cells; Outcome; Pathway interactions; Patients; Peripheral; Pharmacological Treatment; Phenotype; Phosphotransferases; Population; Process; Prognosis; Risk; Role; Sampling; Signal Transduction; Solid Neoplasm; Spatial Distribution; Spleen; T-Lymphocyte; Testing; Translating; Transplantation; Treatment Failure; Tumor Immunity; Tumor Promotion; Tumor-associated macrophages; Up-Regulation; angiogenesis; clinically significant; differential expression; digital; experience; immune checkpoint; immunosuppressive macrophages; in vivo; insight; interest; knock-down; mouse model; mutant; novel; overexpression; peripheral blood; prevent; receptor; single-cell RNA sequencing; transcriptome sequencing; transcriptomics; tumor; tumor growth; tumor microenvironment

Project Summary Low-grade gliomas (LGGs), a heterogeneous group of primary central nervous system tumors, are one of the most common solid tumors in the adolescent and young adult (AYA) population. LGGs can progress to high- grade gliomas (HGGs) via a process known as malignant transformation (MT), resulting in dismal prognoses. The mechanisms driving MT of LGG remain poorly understood. In contrast to LGGs, we and others have shown that HGGs have increased levels of bone marrow-derived myeloid cells (BMDMs) and myeloid-derived suppressor cells (MDSCs) in peripheral circulation, with an intra-tumoral enrichment of tumor-associated macrophages (TAMs) and a paucity of CD8+T and natural killer (NK) cells. Although myeloid cells are known to accumulate during glioma progression, it remains unclear if these cells have a causal role in driving MT. Central hypothesis: Reductions in anti-tumor reactive CD8+T and NK cell-dependent immune surveillance are responsible for LGG to HGG malignant transformation in AYA patients and these reductions are dictated by increased infiltration and immune suppressive activity of tumor infiltrating BMDMs. We have identified two independent myeloid associated pathways that are responsible for these BMDM phenotypes and we propose to test the central hypothesis through the following two aims: Aim 1. Delineate the dependency of BMDM cell differentiation on inhibitor of DNA binding protein 2 (ID2). Using AYA RCAS/tv-a glioma mouse models, we will knockdown ID2 in BMDMs before and during MT. We will examine effects on pro-tumoral myeloid cells and evaluate intra-tumoral T and NK cells and mobilization in blood, spleen, and bone marrow at various times with scRNAseq, mass cytometry, and fluorescent-activated cell sorting. We will also leverage digital spatial transcriptomics (DSP) in ID2 modulated tumors and validate ID2 signaling in human paired LGG /HGG samples to dissect the expression of myeloid, NK, T cell markers, and checkpoint molecules to illuminate ROIs critical to MT. Results will illuminate ID2 mechanisms of BMDM cell differentiation to pro-tumoral myeloid cells. Aim 2. Determine if the CD74/macrophage migration inhibitory factor (MIF) axis regulates BMDM cell differentiation during LGG malignant transformation. We will investigate CD74’s role in MT using pharmacological treatments or transplant wild-type or CD74 KD/KO bone marrow cells in a murine RCAS/tv-a glioma model. We will analyze peripheral blood and the glioma TME using scRNAseq, mass cytometry, and FACS. We will validate findings with bulk RNA sequencing data on paired LGG/HGGs from AYA patients and investigate how CD74/MIF signaling drives reduced T and NK cells which supports LGG MT. We will also leverage DSP in CD74/MIF modulated tumors and validate CD74 signaling in human paired LGG/HGG samples to dissect the expression of myeloid, NK, T cell markers, and checkpoint molecules to illuminate ROIs critical to MT that will indicate potentially targetable vulnerabilities. Overall Impact: Our studies will illuminate novel insights that may enable us to translate into enhanced immune surveillance approaches to delay and/or prevent MT for AYA LGG patients.

项目概要 低级别胶质瘤 (LGG) 是一组异质性原发性中枢神经系统肿瘤，是其中之一 青少年和年轻人 (AYA) 人群中最常见的实体瘤可进展为高危型。 级别胶质瘤（HGG）通过称为恶性转化（MT）的过程，导致预后不良。 与 LGG 相比，我们和其他人已经表明，驱动 LGG MT 的机制仍然知之甚少。 HGG 增加了骨髓源性骨髓细胞 (BMDM) 和骨髓源性骨髓细胞的水平 外周循环中的抑制细胞（MDSC），肿瘤内富集肿瘤相关细胞 巨噬细胞 (TAM) 和缺乏 CD8+T 细胞和自然杀伤 (NK) 细胞，但已知骨髓细胞。 尽管这些细胞在神经胶质瘤进展过程中积累，但尚不清楚这些细胞是否在驱动 MT 中起因果作用。 假设：抗肿瘤反应性 CD8+T 和 NK 细胞依赖性免疫监视的减少是 导致 AYA 患者 LGG 向 HGG 恶性转化，这些减少取决于 我们已经确定了两种肿瘤浸润性 BMDM 的浸润和免疫抑制活性。 独立的骨髓相关途径负责这些 BMDM 表型，我们建议 通过以下两个目标检验中心假设： 目标 1. 描绘 BMDM 细胞的依赖性 使用 AYA RCAS/tv-a 神经胶质瘤小鼠模型对 DNA 结合蛋白 2 (ID2) 抑制剂进行分化。 我们将检查 MT 之前和期间 BMDM 中 ID2 的敲低对促肿瘤骨髓细胞和骨髓细胞的影响。 评估肿瘤内 T 和 NK 细胞以及不同时间在血液、脾脏和骨髓中的动员情况 scRNAseq、质谱流式分析和荧光激活细胞分选我们还将利用数字空间。 ID2 调节肿瘤中的转录组学 (DSP) 并验证人类配对 LGG /HGG 样本中的 ID2 信号传导 剖析骨髓、NK、T 细胞标记物和检查点分子的表达，以阐明对治疗至关重要的 ROI MT。结果将阐明 BMDM 细胞分化为促肿瘤骨髓细胞的 ID2 机制。 确定 CD74/巨噬细胞迁移抑制因子 (MIF) 轴是否调节 BMDM 细胞分化 在 LGG 恶性转化过程中，我们将使用药物治疗研究 CD74 在 MT 中的作用。 或在小鼠 RCAS/tv-a 神经胶质瘤模型中移植野生型或 CD74 KD/KO 骨髓细胞我们将进行分析。 我们将使用 scRNAseq、质谱流式细胞仪和 FACS 来验证外周血和神经胶质瘤 TME 的结果。 来自 AYA 患者的配对 LGG/HGG 的大量 RNA 测序数据，并研究 CD74/MIF 信号如何传导 驱动减少的 T 和 NK 细胞，支持 LGG MT 我们还将在 CD74/MIF 调制中利用 DSP。 肿瘤并验证人类配对 LGG/HGG 样本中的 CD74 信号传导以剖析骨髓的表达， NK、T 细胞标记和检查点分子可照亮对 MT 至关重要的 ROI，这可能表明 总体影响：我们的研究将阐明新的见解，使我们能够转化 强化免疫监测方法，以延迟和/或预防 AYA LGG 患者的 MT。