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.

项目概要/摘要 胶质母细胞瘤（GBM）是一种破坏性脑肿瘤，缺乏有效的治疗方法，这主要是由于胶质母细胞瘤的侵袭。 GBM 细胞能够逃避切除并导致不可避免的复发，通常距离肿瘤 2 厘米 由于存在问题，开发治疗这一过程的进展缓慢。 首先，现有的研究未能认识到这一点。 延伸到肿瘤边缘之外的浸润性 GBM 细胞已经进化出一种独特的适应性细胞机制，这是由于 不幸的是，这些处于侵袭性肿瘤前沿的细胞通常不会受到微环境中的局部应激源的影响。 在分析储存的肿瘤组织的研究中取样的样本，这些组织通常是从容易获得的 另一个问题是大多数侵袭性研究都使用了两种方法。 二维（2D）培养系统涂有一层薄薄的 ECM 蛋白，无法捕获 为了解决这些局限性，我们的团队研究了 GBM 侵袭的维度、机制和异质性。 使用 GBM 定点活检开发了有趣的数据，并拥有组织工程平台可供研究 使用定点活检，我们发现 GBM 细胞侵袭性增加。 外部与内部促进侵袭的整合素和细胞外基质（ECM）剪接变体的表达 我们还从这些定点开发了患者来源的异种移植物（PDX）。 当从肿瘤边缘产生时表现出更大的侵袭性，我们的团队也成为其中之一。 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。 肿瘤周围侵袭下降，基于血管周围整合素的运动增加在这里，我们将基于这一有趣的现象进行研究。 通过研究我们的中心假设来获取数据，即当 GBM 细胞离开肿瘤核心时，与 微环境推动从肿瘤周围侵袭到血管周围侵袭的有针对性的转变。 通过三个目标来实现： 目标 1 – 定义促进肿瘤微环境的变化 肿瘤细胞从 GBM 的中央核心逸出至外缘时发生侵袭性变化；目标 2 - Refine 生物工程培养模型复制 GBM 外缘的微环境并确定其作用 TAM 在驱动该区域的侵袭中​​的作用；目标 3 - 定义整合素在 GBM 侵袭中的作用 细胞从肿瘤外边缘识别并识别该区域的可药物侵袭介质。 为了实现这些目标，我们将使用新型 PDX 和组织工程平台，以及 CRISPRi、单细胞 我们的研究将发现肿瘤细胞和定点活检的新机制。 当细胞迁移远离肿瘤核心时，它们的微环境发生改变，导致侵袭性增加。 这项工作将通过展示 GBM 如何整合不同的区域来挑战传统思维 在确定新的可药物靶点以破坏微环境时，我们将考虑这些区别。 GBM 细胞侵袭性，也可能适用于其他侵袭性癌症。