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.

项目摘要 低级别胶质瘤（LGG）是一组异质性的原发性中枢神经系统肿瘤，是神经系统肿瘤中的一种。 青少年和年轻成人（AYA）人群中最常见的实体瘤。LGG可以发展到高- 恶性胶质瘤（HGG）通过一个称为恶性转化（MT）的过程，导致令人沮丧的脑胶质瘤。 驱动LGG MT的机制仍然知之甚少。与LGG相比，我们和其他人已经表明， HGG的骨髓源性髓样细胞（BMDM）和骨髓源性细胞水平增加 抑制细胞（MDSC）在外周循环中，肿瘤内富集肿瘤相关的 巨噬细胞（TAM）和缺乏CD 8 +T和自然杀伤（NK）细胞。虽然骨髓细胞已知 尽管这些细胞在胶质瘤进展期间积累，但仍不清楚这些细胞是否在驱动MT中具有因果作用。中央 假设：抗肿瘤反应性CD 8 +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，这些减少由以下因素决定： 肿瘤浸润性BMDM的浸润和免疫抑制活性增加。我们已经确定了两个 独立的髓系相关途径，负责这些BMDM表型，我们建议， 通过以下两个目标来检验中心假设：目标1。描述BMDM单元的依赖性 DNA结合蛋白2（ID 2）抑制剂的分化。使用AYA RCAS/tv-a胶质瘤小鼠模型，我们将 在MT之前和期间在BMDM中敲低ID 2。我们将研究对前肿瘤骨髓细胞的影响， 评估肿瘤内T和NK细胞以及血液、脾脏和骨髓中不同时间的动员， scRNAseq、质谱细胞术和荧光激活细胞分选。我们还将利用数字空间 在ID 2调节的肿瘤中使用转录组学（DSP），并在人配对LGG /HGG样品中验证ID 2信号传导 剖析骨髓、NK、T细胞标志物和检查点分子的表达，以阐明对免疫功能至关重要的感兴趣区域。 山结果将阐明BMDM细胞分化为前肿瘤骨髓细胞的ID 2机制。目标2. 确定CD 74/巨噬细胞移动抑制因子（MIF）轴是否调节BMDM细胞分化 LGG恶性转化的过程中。我们将研究CD 74在MT中的作用， 或在鼠RCAS/tv-a神经胶质瘤模型中移植野生型或CD 74 KD/KO骨髓细胞。我们将分析 外周血和胶质瘤TME使用scRNAseq、质谱细胞术和FACS。我们将验证调查结果， AYA患者配对LGG/HGG的批量RNA测序数据，并研究CD 74/MIF信号转导 驱动支持LGG MT的减少的T和NK细胞。我们还将在CD 74/MIF调制中利用DSP 肿瘤中，并验证人配对LGG/HGG样品中的CD 74信号传导， NK、T细胞标志物和检查点分子，以阐明对MT至关重要的ROI， 有针对性的漏洞。总体影响：我们的研究将阐明新的见解，使我们能够翻译 增强免疫监视方法，以延迟和/或预防AYA LGG患者的MT。