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: 9974991
• 负责人: Emmet James Eugene Huang-Hobbs
• 金额: $ 4.27万
• 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/9974991
• 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 Model; 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; 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.

项目总结 成人多形性胶质母细胞瘤(GBM)是最常见和最致命的恶性脑癌。肿瘤 细胞是浸润性的，渗透到周围组织，为复发和逃避治疗提供种子。 GBM的细胞和分子异质性使其治疗进一步复杂化。然而，肿瘤细胞如何 签名在早期肿瘤发生、进展和迁移过程中的变化，以及它们在不同阶段的变化 大脑中的位置仍然未知。此外，侵袭性表型的机制 在这种疾病中观察到的情况知之甚少。因此，这项提案的首要目标是界定如何 随着时间的推移，肿瘤细胞会侵入周围组织，并确定推动这一过程的机制。 最近对入侵的GBM前沿进行抽样的研究发现，在毫米范围内有不同的种群 说明了在分析肿瘤细胞功能时微环境的重要性。要映射 随着时间的推移，GBM在大脑不同区域的演变，我们发展了一种内源性 免疫活性小鼠模型，使我们能够研究自然肿瘤背景下的细胞侵袭。我们的预赛 数据表明，肿瘤细胞的表达随肿瘤位置的不同而不同，并与功能相关；然而，当 而在疾病发展过程中，这些变化发生在哪里仍不清楚。因此，该计划的首要目标是 建议的研究是确定侵袭的动力学以及相关基因和蛋白的表达变化。 在我们的天然肿瘤模型的背景下。我们将使用肿瘤的3D映射和RNA-Seq研究 在多个时间点肿瘤中三个不同的位置(原发肿瘤、继发肿瘤、胼胝体) 以明确基底膜的侵袭动态。 在初步研究中，我们发现轴突引导基因在继发性肿瘤部位和体部丰富。 老茧。因为它们在中枢神经系统内的迁移中起着既定的作用，特别是在 对于癌症，我们进行了轴突引导基因的编码功能(GOF)筛查。我们发现EphA7 在原发灶外特异性地浓集，提示它有助于基底膜的迁移。而EphA7 在GBM患者中，表达与较差的预后相关，其在侵袭中的作用尚不清楚。因此，我们的第二个 目的：通过对肾小管上皮细胞功能的获得和丧失的研究，明确EphA7在基底膜侵袭和迁移中的作用 我们的天然肿瘤模型和人类患者来源的异种移植的背景。 总体而言，这项建议旨在扩展我们的初步数据，以生成完整的时间表和3D地图 在肿瘤起始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本地肿瘤模型中验证这些更改，然后突出显示 在体外和体内与人类系统相似。