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 低度神经胶质瘤（LGGS）是一组异质的原发性中枢神经系统肿瘤，是其中之一 青少年和年轻人（AYA）人口中最常见的实体瘤。 LGG可以发展到高 级胶质瘤（HGGS）通过称为恶性转化（MT）的过程，导致惨淡的进展。 驱动LGG的MT的机制仍然鲜为人知。与LGG相反，我们和其他人都表明 HGG的骨髓衍生的髓样细胞（BMDM）和髓样衍生的水平增加 抑制细胞（MDSC）在外周循环中，肿瘤相关的肿瘤内酶 巨噬细胞（TAM）和CD8+T和天然杀手（NK）细胞的稀少。尽管已知髓样细胞 在神经胶质瘤进展过程中积累，尚不清楚这些细胞在驱动MT中是否具有因果作用。中央 假设：抗肿瘤反应性CD8+T和NK细胞依赖性免疫保护率的减少是 负责AYA患者的LGG到HGG恶性转化，这些降低由 肿瘤浸润BMDM的浸润和免疫抑制活性增加。我们已经确定了两个 负责这些BMDM表型的独立髓样相关途径，我们建议 通过以下两个目的测试中心假设：目标1。描述BMDM单元的依赖性 DNA结合蛋白2的抑制剂分化（ID2）。使用AYA RCAS/TV-A Glioma鼠标模型，我们将 MT之前和期间BMDMS中的ID2敲低ID2。我们将检查对肌肌髓髓细胞的影响和 在不同时间评估肿瘤内T和NK细胞的血液，脾和骨髓的动员 SCRNASEQ，质量细胞仪和荧光激活的细胞分选。我们还将利用数字空间 ID2调制肿瘤中的转录组学（DSP），并在人配对LGG /HGG样品中验证ID2信号传导 剖析髓样，NK，T细胞标记物和检查点分子的表达，以照明对ROI至关重要的ROI 公吨。结果将阐明BMDM细胞分化与肌髓髓样细胞的ID2机制。目标2。 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恶性转化期间。我们将使用药物治疗研究CD74在MT中的作用 或在鼠RCAS/TV-A神经瘤模型中的移植野生型或CD74 KD/KO骨髓细胞。我们将分析 使用SCRNASEQ，质量细胞术和FACS，外周血和神经胶质瘤TME。我们将通过 来自AYA患者配对的LGG/HGG的大量RNA测序数据，并研究CD74/MIF信号如何传导 驱动降低的T和NK细胞支持LGG MT。我们还将利用CD74/MIF调制的DSP 肿瘤和验证人配对的LGG/HGG样品中的CD74信号传导以剖析髓样的表达， NK，T细胞标记物和检查点分子点亮对MT至关重要的ROI，这可能表明可能 可目标漏洞。总体影响：我们的研究将阐明新颖的见解，使我们能够翻译 进入增强的免疫外科手术方法，以延迟和/或预防AYA LGG患者MT。