Biophysical Interrogation of Signals that Drive GBM Invasion

驱动 GBM 侵袭的信号的生物物理询问

基本信息

批准号：
10449770
负责人：
Adam J Engler
金额：
$ 6.14万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10449770
关键词：
ATAC-seq Address Adhesions Adhesives Adult Affect Astrocytes Behavior Biochemical Biological Assay Biological Models Biophysics Brain Cell Adhesion Cells ChIP-seq Chromatin Clinical Co-Immunoprecipitations Coculture Techniques Color Combined Modality Therapy Complex Contralateral Data Dependence Disease Distal Education Enhancers Environment Epidermal Growth Factor Receptor Epigenetic Process Excision Exons Exposure to Extracellular Matrix Extracellular Matrix Proteins Fibronectins Fluorescence-Activated Cell Sorting Focal Adhesions Fostering Gene Expression Gene Expression Profile Gene Silencing Genes Genetic Genetic Diseases Genetic Engineering Genetic Heterogeneity Genotype Glioblastoma Glioma Heterogeneity High-Throughput Nucleotide Sequencing Histologic Human In Vitro Injections Integrins Intervention Invaded Lead Lesion Malignant Glioma Measurement Mediating Methods Modeling Molecular Mus Mutate Mutation Neurologic Operative Surgical Procedures Pathogenicity Pathway Analysis Pathway interactions Patients Pharmacology Phenotype Phosphotransferases Phosphotyrosine Population Primary Brain Neoplasms Process Production Prognosis Proteins Radiation Receptor Protein-Tyrosine Kinases Role Sampling Serial Passage Signal Pathway Signal Transduction Signaling Protein Sorting - Cell Movement Stains System Technology Testing Time Tyrosine Kinase Inhibitor Variant adhesion receptor base brain parenchyma chemotherapy comparative cytokine disease prognosis epidermal growth factor receptor VIII extracellular improved in vivo induced pluripotent stem cell migration mutant neoplastic cell novel therapeutic intervention paracrine promoter public database receptor recruit transcriptome sequencing tumor

项目摘要

One major issue confounding successful treatment of glioblastoma multiforme (GBM) is the presence of highly invasive cells disseminating into the brain parenchyma. These cells evade surgical resection and often spread distally in brain parenchyma. Multiple and spatially distinct heterotypic populations exist within a single GBM, giving rise to the disease’s genetic heterogeneity and leading to complex cell intrinsic and extrinsic mechanisms of invasion. Amplification of the epidermal growth factor receptor (EGFR), a hallmark mutation present in 60% of cases, most often occurs in a heterogeneous manner and is frequently associated with deletion of exons 2-7, creating a constitutively active mutant, EGFRvIII. While significant focus has been placed on its kinase activity, comparatively little is known about EGFRvIII’s ability to enhance migration via interaction with adhesion receptors. Our preliminary data supports a dual role for EGFRvIII where it interferes with intrinsic adhesion receptors and also recruits non-transformed counterparts via extrinsic signaling to reduce adhesion of a mixed population. Based on our findings, we hypothesize that this difference in adhesive activity is due to differential signaling associated with EGFRvIII, and that this receptor conveys this phenotype to non-transformed counterparts through cytokine production (Inda, Genes & Dev, 2010; Zanca, Genes & Dev, 2017) to cooperatively invade parenchyma. With this hypothesis, we will use adhesion measurement technologies to dissect cell intrinsic EGFR-mediated invasion mechanisms; given the heterogeneity within tumors, we will also combine newly developed adhesion sorting technologies with high throughput sequencing technologies to identify cell extrinsic mechanisms and targets for subsequent intervention. The following lines of experimentation will be carried out: 1) implementation of biophysical assays and signaling pathway analyses to interrogate how cell intrinsic activity of EGFRvIII leads to labile adhesion and an invasive phenotype; 2) biochemical and functional analysis of the EGFRvIII cell extrinsic, secretome-mediated education of wtEGFR cell adhesive phenotype; 3) expression and epigenetic analyses on adhesion-sorted populations will be used to define a migratome signature, its stability in wtEGFR cells after exposure to the EGFRvIII secretome, and the ability of “educated” wtEGFR to propagate that epigenetic signature to naïve wtEGFR cells.

阻碍多形性胶质母细胞瘤 (GBM) 成功治疗的一个主要问题是存在高度侵袭性细胞播散到脑实质中，这些细胞逃避手术切除并经常扩散。在脑实质的远端，单个 GBM 内存在多个且空间上不同的异型群体，引起疾病的遗传异质性并导致复杂的细胞内在和外在表皮生长因子受体（EGFR）的扩增，这是一种标志性突变。出现在 60% 的病例中，最常以异质方式发生，并且经常与外显子 2-7 的缺失，产生了一个组成型活性突变体 EGFRvIII。就其激酶活性而言，人们对 EGFRvIII 通过以下方式增强迁移的能力知之甚少：我们的初步数据支持 EGFRvIII 具有干扰作用的双重作用。具有内在粘附受体，并且还通过外在信号传导招募未转化的供体根据我们的研究结果，我们发现粘合剂的这种差异。活性是由于与 EGFRvIII 相关的差异信号传导，并且该受体传达这种表型通过细胞因子产生转化为非转化单位（Inda，Genes & Dev，2010；Zanca，Genes & Dev， 2017）合作侵入薄壁组织根据这个假设，我们将使用粘附测量。考虑到内部的异质性，剖析细胞内在 EGFR 介导的侵袭机制的技术；肿瘤，我们还将把新开发的粘附分选技术与高通量测序相结合识别细胞外在机制和后续干预目标的技术。将进行的实验：1）实施生物物理测定和信号通路分析探究 EGFRvIII 的细胞内在活性如何导致不稳定的粘附和侵袭性表型 2) EGFRvIII 细胞的生化和功能分析 wtEGFR 的外在、分泌组介导的教育细胞粘附表型；3) 将使用粘附分选群体的表达和表观遗传学分析定义迁移组特征、其在暴露于 EGFRvIII 分泌组后在 wtEGFR 细胞中的稳定性，以及 “受过教育的”wtEGFR 将表观遗传特征传播到初始 wtEGFR 细胞的能力。