Tissue-Engineered Models of Glioblastoma for Evaluating Treatment Response

胶质母细胞瘤(GBM)是一种对治疗反应差且几乎不可避免的致命性脑癌。 复发。基底膜肿瘤不会转移。相反，它们会积极地渗透到大脑中。肿瘤边界是 作为促进GBM细胞存活的关键利基环境的微血管高度血管化， 治疗耐药和迁移。目前，人们认为一个耐治疗的干细胞亚群- 像GBM细胞(GSCs)一样，从原发肿瘤迁出--与局部微血管保持密切联系 -和种子复发肿瘤-这些肿瘤富含GSC。然而，这种影响背后的机制 基底膜细胞移离原发肿瘤，导致复发的微血管生态位在很大程度上仍然存在 未知。为了解决这一知识差距，拟议的研究将调查1)特定线索在 肿瘤微环境对肾小球系膜细胞迁移表型的影响及2)这些信号对细胞迁移表型的影响 异质肿瘤内不同的细胞亚群，包括GSCs。我们的基本假设是 肿瘤微血管周围的微环境通过两种机械途径促进GBM进展 以及化学手段。目标1将考察机械线索的效应，目标2将考察生化线索的效应 存在于GBM迁移表型的微血管壁龛中。为了做到这一点，我们将使用一种创新的组织- 用于3D培养患者来源的GBM细胞的工程平台，以量化如何特定的特征 微环境影响迁徙行为。我们建议模块化地调整机械和化学性能 培养平台，以确定其对GBM迁移表型的独立影响。对一些人来说 实验，平台将在定义的微环境之间建立接口，通过该接口 GBM细胞可以迁移，使迁移和非迁移细胞能够分离，以便进行下游分析和 提供关于单个肿瘤内的细胞亚群，如GSCs如何反应的信息 与微环境线索不同。由于治疗耐药，侵袭和复发是相互关联的。 事件，我们将调查暴露在常规临床治疗下如何改变GBM的迁移反应 微环境线索。利用单细胞RNA测序，我们将研究潜在的机制 对微血管壁龛的特定特征的反应。总的来说，拟议的研究将使 我们对GBM与微血管壁龛的相互作用如何促进肿瘤的理解取得了进展 复发的特征：1)基底膜肿瘤和微血管的生物力学特性(目标1)，2)迁移性 GBM细胞对生化和生物力学提示的反应(目标1和2)和3)支配机制 对生化和生物力学提示的迁移反应(目标1和2)。从长远来看，我们预计 这些见解将导致针对GBM复发的新疗法的开发。 好了！

Glioblastoma (GBM) is a uniformly fatal brain cancer with poor response to treatment and virtually inevitable recurrence. GBM tumors do not metastasize. Instead, they aggressively infiltrate the brain. Tumor borders are highly vascularized by microvessels that serve as key niche environments promoting GBM cell survival, treatment resistance and migration. Currently, it is believed that a subpopulation of treatment-resistant, stem- like GBM cells (GSCs) migrate away from primary tumors — remaining in close contact with local microvessels — and seed recurrent tumors — which are enriched in GSCs. However, the mechanisms underlying the influence of the microvessel niche on GBM cell migration away from primary tumors, leading to recurrence, remain largely unknown. To address this gap in knowledge, the proposed studies will investigate 1) the role of specific cues in the tumor microenvironment on the migratory phenotype of GBM cells and 2) how these cues differentially affect distinct subpopulations of cells within heterogeneous tumors, including GSCs. Our fundamental hypothesis is that the microenvironment surrounding tumor microvessels promotes GBM progression through both mechanical and chemical means. Aim 1 will investigate effects of mechanical cues and Aim 2 effects of biochemical cues present in the microvessel niche on migratory phenotype of GBM. To do this, we will use an innovative, tissue- engineered platform for 3D culture of patient-derived GBM cells to quantify how specific features of the microenvironment affect migratory behavior. We propose to modularly tune mechanical and chemical properties of culture platforms to characterize their independent effects on migratory phenotype of GBM. For some experiments, platforms will be constructed with an interface between defined microenvironments through which GBM cells can migrate, enabling separation of migratory and non-migratory cells for downstream analysis and providing information about how subpopulations of cells within a single tumor, such as GSCs, may respond differentially to microenvironmental cues. As treatment resistance, invasion and recurrence are inter-related events, we will investigate how exposure to routine clinical therapies may alter migratory responses of GBM to microenvironmental cues. Using single-cell RNA sequencing, we will investigate the mechanisms underlying responses to specific features of the microvessel niche. Collectively, the proposed studies will make significant strides towards our understanding of how interactions of GBM with the microvessel niche promote tumor recurrence by characterizing: 1) biomechanical properties of GBM tumors and microvessels (Aim 1), 2) migratory response of GBM cells to biochemical and biomechanical cues (Aims 1 and 2) and 3) mechanisms governing the migratory responses to biochemical and biomechanical cues (Aims 1 and 2). In the long term, we expect these insights will lead to the development of new therapies targeting GBM recurrence. !