Modeling and druggable-genome screening of glioblastoma invasion using regional biopsy-guided biomaterials systems

使用区域活检引导的生物材料系统对胶质母细胞瘤侵袭进行建模和药物基因组筛选

• 批准号: 10237253
• 负责人: Manish Aghi
• 金额: $ 41.4万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-17 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10237253
• 关键词: 3-Dimensional; Address; Automobile Driving; Biocompatible Materials; Biological Models; Biomedical Engineering; Biopsy; Blood capillaries; Brain Neoplasms; CD44 gene; CRISPR interference; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen Type IV; Data; Diagnosis; Dimensions; Engineering; Excision; Exhibits; Extracellular Matrix Proteins; Failure; Fibronectins; Generations; Genome; Glioblastoma; Goals; Heterogeneity; Hyaluronic Acid; Hydrogels; In Vitro; Integrins; Knock-out; Laminin; Libraries; Ligands; Location; Magnetic Resonance Imaging; Malignant Neoplasms; Malignant neoplasm of brain; Mechanics; Mediating; Mediator of activation protein; Methods; Modality; Modeling; Nuclear; Patients; Process; Prognosis; Property; RNA Splicing; Recurrence; Role; Route; Sampling; Site; Structure; System; Technology; Thinking; Thinness; Tissue Engineering; Tumor Bank; Tumor Tissue; Tumor-associated macrophages; Variant; Work; base; cancer invasiveness; cell motility; combat; druggable target; effective therapy; falls; gray matter; high throughput screening; in vivo; loss of function; neoplastic cell; new therapeutic target; novel; patient derived xenograft model; screening; single cell technology; stressor; therapy development; three-dimensional modeling; tumor; tumor microenvironment; two-dimensional; white matter

PROJECT SUMMARY/ABSTRACT Glioblastoma (GBM) is a devastating brain tumor lacking effective treatments. This is largely due to invasion of GBM cells, which enables escape from resection and drives inevitable recurrence, typically 2 cm from the location at diagnosis. Progress in developing therapies to combat this process has been slow due to problems with the cells being studied and the methods of analysis. First, existing studies have failed to recognize that infiltrating GBM cells extending beyond the tumor edge have evolved a unique adaptive cellular machinery due to local stressors in their microenvironment. Unfortunately, these cells at the invasive tumor front are often not the ones sampled in studies analyzing banked tumor tissue, which is typically procured from the readily accessible central portions of the tumor. Another problem is that most studies of invasiveness have used two- dimensional (2D) culture systems coated with a thin layer of ECM proteins, which fail to capture the dimensionality, mechanics, and heterogeneity of GBM invasion. To address these limitations, our team has developed intriguing data using site-directed biopsies from GBM and has tissue engineered platforms to study invasion in vitro. Using site-directed biopsies, we have shown increased GBM cell invasiveness and increased expression of invasion-promoting integrins and extracellular matrix (ECM) splice variants outside versus inside enhancing MRI regions. We have also developed patient-derived xenografts (PDXs) from these site-directed biopsies that exhibit more invasiveness when arising from the tumor edge. Our team also became among the first to bioengineer 3D hydrogel systems as a discovery platform In GBM. We found that, as CD44-mediated peritumoral invasion falls, perivascular integrin-based motility increases. Here, we will build upon this intriguing data by investigating our central hypothesis that as GBM cells exit the tumor core, reciprocal interactions with the microenvironment drive a targetable transition from peritumoral to perivascular invasion. These goals will be accomplished through three aims: Aim 1 – Define changes in the tumor microenvironment promoting invasive change as tumor cells egress away from the central core to the outer edge of GBM; Aim 2 - Refine bioengineered culture models to replicate the microenvironment of the outer edge of GBM and identify the role of TAMs in driving invasion in this region; and Aim 3 - Define the role of integrins in the invasiveness of GBM cells from the outer tumor edge and identify druggable mediators of invasion in this region. To accomplish these goals, we will use novel PDXs and tissue engineered platforms, along with CRISPRi, single-cell technology, and site-directed biopsies. Our studies will discover novel mechanisms by which tumor cells and their microenvironment are altered to drive increased invasiveness as cells migrate away from the tumor core. This work will challenge conventional thinking by showing how GBM integrates distinct regional microenvironments. We will account for these distinctions when identifying novel druggable targets to disrupt GBM cell invasiveness, with potential applicability to other invasive cancers as well.

项目摘要/摘要 胶质母细胞瘤（GBM）是缺乏有效治疗的毁灭性脑肿瘤。这主要是由于入侵 GBM细胞，可以逃脱切除并不可避免地复发，通常从距离 诊断处的位置。由于问题而开发打击这一过程的疗法的进展一直很慢。 随着细胞的研究和分析方法。首先，现有研究未能意识到 渗透到肿瘤边缘超出肿瘤边缘的GBM细胞已发展出独特的自适应细胞机械。 在其微环境中向当地压力源。不幸的是，这些细胞在侵入性肿瘤方面通常不是 在分析银行肿瘤组织的研究中取样的研究，通常是从该组织中获取的 肿瘤的无障碍中央部分。另一个问题是，大多数侵入性研究都使用了两种 涂有薄层ECM蛋白的尺寸（2D）培养系统，无法捕获 GBM入侵的维度，力学和异质性。为了解决这些限制，我们的团队有 使用来自GBM的站点定向的活检开发了有趣的数据，并具有组织工程平台来研究 体外入侵。使用定向的活检，我们显示GBM细胞的侵入性增加并增加 在外部与内部的侵袭性整合素和细胞外基质（ECM）剪接变体的表达 增强MRI区域。我们还从这些站点定向的 从肿瘤边缘产生时暴露出更多侵入性的活检。我们的团队也成为 首先将生物工程3D水凝胶系统作为GBM的发现平台。我们发现，作为CD44介导的 周围侵袭下降，基于血管内整联蛋白的运动增加。在这里，我们将基于这个有趣的 通过研究我们的中心假设来数据，当GBM细胞退出肿瘤核心时，与之相互作用 微环境驱动从周围到血管周围侵袭的目标过渡。这些目标将 通过三个目标完成：目标1 - 定义促进肿瘤微环境的变化 当肿瘤细胞从中心核心流入GBM的外边缘时，侵入性变化；目标2-精炼 生物工程培养模型以复制GBM外边缘的微环境并确定角色 在该地区驾驶入侵的TAM；目标3-定义整联蛋白在GBM侵入性中的作用 来自外部肿瘤边缘的细胞，并鉴定该区域的可药物浸润介质。完成 这些目标，我们将使用新颖的PDX和组织工程平台，以及CRISPRI，单细胞 技术和现场指导的活检。我们的研究将发现肿瘤细胞和 随着细胞从肿瘤核心迁移，它们的微环境改变了以提高侵入性。 这项工作将通过展示GBM如何整合不同的区域来挑战常规思维 微环境。在识别新型可药物目标破坏时，我们将考虑这些区别 GBM细胞的侵入性，也可能适用于其他侵入性癌症。