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：
10819632
负责人：
Adam J Engler
金额：
$ 2.43万
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/10819632
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 Engineering Genetic Heterogeneity Genotype Glioblastoma Glioma Heterogeneity High-Throughput Nucleotide Sequencing Histologic Human In Vitro Injections Integrins Intervention Invaded Lesion Malignant Glioma Measurement Mediating Methods Modeling Molecular Mus Mutate Mutation Neurologic Operative Surgical Procedures Pathogenicity Pathway Analysis Pathway interactions Patients Phenotype Phosphotransferases Phosphotyrosine Population Primary Brain Neoplasms Process Production Prognosis Proteins Radiation therapy Receptor Cell Receptor Protein-Tyrosine Kinases Recurrence Role Sampling Serial Passage Signal Pathway Signal Transduction Sorting Stains System Technology Testing Time Tyrosine Kinase Inhibitor Variant adhesion receptor 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 pharmacologic promoter public database receptor recruit transcriptome sequencing transmission process 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.

一个主要的问题，即成功治疗多形胶质母细胞瘤（GBM）是高度的存在这些细胞逃避手术切除并经常扩散在脑实质方面的不同。多个和空间上不同的异型种群存在于单个GBM中，引起该疾病的遗传异质性，并导致复杂的细胞固有和外在入侵机制。表皮生长因子受体（EGFR）的扩增，标志性突变在60％的情况下存在，大多数情况通常以异质方式发生，并且经常与删除外显子2-7，创建一个组成型活跃的突变体Egfrviii。虽然重点是在其激酶活性上，关于EGFRVIII通过与粘合剂受体的相互作用。我们的初步数据支持EGFRVIII相互作用的双重作用具有固有的粘合剂受体，还通过外部信号传导招募非转化的对应物减少种群混合的粘合剂。根据我们的发现，我们假设这种粘合剂的差异活动是由于与EGFRVIII相关的差异信号引起的，并且该接收器传达了此表型通过细胞因子产生非转化的对应物（Inda，Genes＆Dev，2010； Zanca，Genes＆Dev， 2017年）合作入侵Paranchyma。有了这个假设，我们将使用粘合剂测量剖析细胞内在的EGFR介导的侵袭机制的技术；鉴于内部的异质性肿瘤，我们还将结合新开发的粘附分选技术与高吞吐量测序鉴定细胞外在机制和随后干预的靶标的技术。以下行实验将进行：1）实施生物物理测定和信号通路分析询问EGFRVIII的细胞内在活性如何导致标签粘附和侵入性表型； 2） EGFRVIII细胞外部，分泌介导的WTEGFR教育的生化和功能分析细胞粘合剂表型； 3）将使用对粘合剂分类种群的表达和表观遗传分析为了定义一个迁移符号，其在暴露于EGFRVIII分泌组的WTEGFR细胞中的稳定性，并且 “受过教育” WTEGFR传播这种表观遗传学签名到幼稚的WTEGFR细胞的能力。