Defining invasion dynamics of GBM reveals axon guidance genes drive invasion

定义 GBM 的侵袭动力学揭示轴突引导基因驱动侵袭

• 批准号: 10219201
• 负责人: Emmet James Eugene Huang-Hobbs
• 金额: $ 4.32万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10219201
• 关键词: 3-Dimensional; Adult; Adult Glioblastoma; Appearance; Automobile Driving; Bar Codes; Behavior; Biological Assay; Brain; Brain region; CRISPR/Cas technology; Cell physiology; Cells; Chemotaxis; Chemotherapy and/or radiation; Contralateral; Corpus Callosum; Data; Diagnosis; Disease; Disease Progression; Electroporation; Environment; Event; Evolution; Gene Expression; Gene Expression Profile; Genes; Genetic; Glioblastoma; Goals; Heterogeneity; Human; Immunocompetent; Immunocompromised Host; Immunohistochemistry; In Vitro; Infiltration; Injections; Invaded; Knowledge; Light; Location; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Maps; Measures; Microscopy; Modeling; Molecular; Mus; Neoplasm Metastasis; Neuraxis; Operative Surgical Procedures; Outcome; Patients; Pattern; Phenotype; Population; Population Heterogeneity; Primary Neoplasm; Process; Proteins; Recurrence; Regimen; Relapse; Role; Sampling; Sampling Studies; Seeds; Site; Survival Rate; System; Testing; Time; TimeLine; Tissue imaging; Tissues; Tumor Cell Invasion; Tumor Cell Migration; Up-Regulation; Xenograft procedure; axon guidance; base; cancer cell; gain of function; in utero; in vitro Model; in vivo; in vivo Model; insight; loss of function; migration; millimeter; mouse model; neoplastic cell; novel strategies; overexpression; patient derived xenograft model; protein expression; spatial relationship; transcriptome sequencing; tumor; tumor heterogeneity; tumor initiation; tumor progression; tumorigenesis; virtual

PROJECT SUMMARY Adult glioblastoma multiforme (GBM) is the most common and deadly form of malignant brain cancer. Tumor cells are infiltrative, permeating surrounding tissue, providing the seeds for recurrence, and evading therapy. The cellular and molecular heterogeneity of GBM further complicate its treatment. However, how tumor cell signatures change during early tumorigenesis, progression, and migration and how they vary at different locations in the brain remain unknown. Additionally, the mechanisms responsible for the invasive phenotype observed in this disease are poorly understood. Therefore, the overarching goal of this proposal is to define how tumor cells invade surrounding tissues over time and identify the mechanism driving this process. Recent studies sampling the leading edges of invading GBM have found diverse populations within millimeters of one another, illustrating the importance of microenvironment when analyzing tumor cell function. To map the evolution of GBM through different regions of the brain over time, we developed an endogenous immunocompetent mouse model that allows us to study cell invasion in a native tumor context. Our preliminary data indicate that tumor cell expression varies by tumor location and correlates with function; however, when and where these changes arise during disease progression remains unknown. Therefore, the first aim of the proposed study is to define the dynamics of invasion and associated gene and protein expression changes in GBM within the context of our native tumor model. We will use 3D mapping of the tumor and RNA-Seq studies of three distinct locations (primary tumor, secondary tumor, corpus callosum) in the tumor at multiple time points in order to define the invasion dynamics of GBM. In preliminary studies, we found axon guidance genes to be enriched in the secondary tumor site and corpus callosum. Because of their established role in migration within the central nervous system and specifically in cancer, we performed a barcoded gain of function (GOF) screen of axon guidance genes. We found that EphA7 was specifically enriched outside of the primary tumor, suggesting it contributes to GBM migration. While EphA7 expression is associated with worse outcomes in GBM patients, its role in invasion is unknown. Thus, our second aim is to define the role of EphA7 in GBM invasion and migration through gain and loss of function studies in the context of our native tumor model and in human patient derived xenografts. Overall, this proposal aims to expand upon our preliminary data to generate a complete timeline and 3D map of the genetic events and corresponding expression changes that occur between tumor initiation and invasion through the brain. We will then validate those changes first in our native tumor model and then will highlight the parallels with human systems in vitro and in vivo.

项目概要 成人多形性胶质母细胞瘤（GBM）是最常见和致命的恶性脑癌。瘤 细胞是浸润性的，渗透到周围组织，为复发提供种子并逃避治疗。 GBM 的细胞和分子异质性进一步使其治疗变得复杂。然而，肿瘤细胞如何 特征在早期肿瘤发生、进展和迁移过程中发生变化，以及它们在不同时期的变化方式 大脑中的位置仍然未知。此外，造成侵袭表型的机制 人们对这种疾病的观察知之甚少。因此，该提案的总体目标是定义如何 随着时间的推移，肿瘤细胞侵入周围组织并确定驱动这一过程的机制。 最近对入侵 GBM 前沿进行采样的研究发现，在毫米范围内存在不同的种群 一方面，说明了微环境在分析肿瘤细胞功能时的重要性。绘制地图 随着时间的推移，GBM 通过大脑不同区域的进化，我们开发了一种内源性 免疫活性小鼠模型使我们能够研究天然肿瘤环境中的细胞侵袭。我们的初步 数据表明，肿瘤细胞的表达随肿瘤位置的不同而变化，并与功能相关；然而，当 在疾病进展过程中这些变化是在哪里发生的仍然未知。因此，该计划的首要目标是 拟议的研究是为了定义入侵的动态以及相关基因和蛋白质表达的变化 GBM 在我们的天然肿瘤模型的背景下。我们将使用肿瘤的 3D 绘图和 RNA 测序研究 多个时间点肿瘤中三个不同位置（原发肿瘤、继发肿瘤、胼胝体）的 为了定义 GBM 的侵袭动力学。 在初步研究中，我们发现轴突导向基因在继发性肿瘤部位和语料库中富集 胼胝体。由于它们在中枢神经系统内迁移中的既定作用，特别是在 癌症方面，我们对轴突引导基因进行了条形码功能获得（GOF）筛选。我们发现EphA7 在原发肿瘤之外特别富集，表明它有助于 GBM 迁移。而EphA7 表达与 GBM 患者的较差预后相关，但其在侵袭中的作用尚不清楚。因此，我们的第二个 目的是通过功能获得和丧失的研究来确定 EphA7 在 GBM 侵袭和迁移中的作用 我们的天然肿瘤模型和人类患者衍生的异种移植物的背景。 总体而言，该提案旨在扩展我们的初步数据以生成完整的时间线和 3D 地图 肿瘤发生和侵袭之间发生的遗传事件和相应的表达变化 通过大脑。然后，我们将首先在我们的天然肿瘤模型中验证这些变化，然后突出显示 与体外和体内的人体系统相似。