Crosstalk between EGFR and TEAD Activity Directs Migration in Human Glioblastoma

EGFR 和 TEAD 活性之间的串扰指导人胶质母细胞瘤的迁移

基本信息

批准号：
10079757
负责人：
Nadejda Mincheva Tsankova
金额：
$ 1.1万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-15 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10079757
关键词：
ATAC-seq Adhesions Affect Automobile Driving Behavior Binding Biochemical Biology Brain Brain Neoplasms CRISPR/Cas technology Carcinoma Cell Culture Techniques Cells ChIP-seq Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Complex DNA Data Dependence Diffuse Disease Progression Down-Regulation Elements Engraftment Epidermal Growth Factor Receptor Epigenetic Process Equipment Excision Extracellular Matrix FDA approved Family Feedback Funding Gene Expression Gene Targeting Genes Genetic Transcription Genomics Glioblastoma Glioma Goals Growth Histology Human Hypoxia Immunocompetent Immunofluorescence Immunologic In Vitro Infiltration Infiltrative Growth Knock-in Knock-out Libraries Link Malignant Neoplasms Measures Mediating Metabolic Metabolism Methods Modeling Molecular Morphologic artifacts Mus Nucleic Acid Regulatory Sequences Oncogenic Operative Surgical Procedures PDGFRA gene Pathway interactions Patients Penetrance Pharmacology Phenotype Preparation Proto-Oncogene Proteins c-akt RNA Recurrence Role SCID Mice Signal Transduction Slice Solid Neoplasm System Testing Time Toxic effect Treatment Efficacy Tumor Burden Tumor Cell Biology Verteporfin Xenograft procedure base cell growth cell motility chemotherapy cost experimental study fluorescence microscope genome-wide in vivo inhibitor/antagonist insight migration neoplastic cell neuroinflammation novel novel therapeutics overexpression prevent progenitor programs qubit stem cell population stressor synergism transcription factor transcriptome sequencing tumor tumor growth tumor progression

项目摘要

SUMMARY Glioblastoma (GBM) tumors manifest with a large, proliferative core and a diffusely infiltrative border, the latter contributing to incomplete surgical resection and inevitable tumor recurrence. A better understanding of the specific molecular pathways responsible for cell infiltration in GBM may thus provide novel and potentially more effective opportunities for therapy. The biology of tumor cell infiltration/migration is complex and intimately linked to its microenvironment, including adaptations for unique metabolic, hypoxic, motility, extracellular matrix, and neuroinflammatory stressors. The pathways are difficult to recapitulate in glioma models that rely on cell culturing. As cell fate decisions converge on the level of epigenetic control, one attractive strategy to study the intricate biology of migration is to use epigenetics to define the transcriptional regulators of migration in a system that closely resembles the native tumor cell state. In this proposal, we take advantage of our lab's epigenetic expertise to study cell fate states in glioma stem cell populations (GSCs), directly isolated from their tumor niche without pre-culture artifact [37-38,57]. Through the unique comparison of open chromatin in freshly derived GSCs and normal germinal matrix progenitors, we were able to distinguish the transcriptionally accessible regions in GSCs that specifically relate to cell migration (in final revisions). This allowed us to infer the most salient transcription factor (TF) motifs within tumor-specific regulatory regions linked to migration, where the TEAD family emerged as a top candidate. The TEAD TFs, along with their co- activators YAP/TAZ, are the main effectors of the Hippo pathway, and have been studied for their oncogenic regulatory role predominantly outside of the brain. To test the functional role of YAP-TEAD in GBM growth and migration, we generated TEAD1 knockout in patient-derived GSCs using CRISPR/Cas9 and employed the YAP-TEAD inhibitor, Verteporfin, to inhibit TEAD activity. Using both methods to inhibit TEAD1 activity, we detected reduced capacity for cell migration in vitro and robust downregulation of EGFR and pERK expression. EGFR knockout similarly displayed reduced migration in vitro, as well as decreased TEAD activity. Based on this, here we hypothesize that TEAD activity regulates infiltrative growth in GBM, mediated at least partially through an EGFR regulatory loop. To test this hypothesis, we aim to 1) systematically define TEAD occupancy at open chromatin and downstream TEAD-target genes across GBM subtypes; 2a) dissect mechanistically the relationship between Hippo-YAP/TAZ-TEAD and EGFR/RTK-ERK; 2b) define the role of TEAD1 in regulating migration and growth in vitro and in/ex vivo, and its synergy with EGFR; and 3) test the potential therapeutic efficacy of YAP-TEAD inhibitors, such as Verteporfin. We envision YAP/TAZ-TEAD as an important regulator of infiltrative growth in GBM, where pharmacologic inhibition of its activity alleviates tumor burden while also blocking oncogenic RTK effects, thus offering new therapeutic options for patients with GBM.

概括胶质母细胞瘤 (GBM) 肿瘤表现为大的增殖性核心和弥漫性浸润性边界，后者导致手术切除不完全和不可避免的肿瘤复发。更好地理解因此，负责 GBM 细胞浸润的特定分子途径可能提供新颖且可能更多的信息。有效的治疗机会。肿瘤细胞浸润/迁移的生物学复杂且密切相关与其微环境有关，包括对独特的代谢、缺氧、运动、细胞外的适应基质和神经炎症应激源。这些途径很难在依赖于神经胶质瘤模型中重现关于细胞培养。随着细胞命运决定集中在表观遗传控制水平上，一种有吸引力的策略研究迁徙的复杂生物学就是利用表观遗传学来定义迁徙的转录调节因子在与天然肿瘤细胞状态非常相似的系统中迁移。在这个提案中，我们利用我们实验室的表观遗传学专业知识可直接研究神经胶质瘤干细胞群 (GSC) 的细胞命运状态从肿瘤生态位中分离出来，无需预培养工件[37-38,57]。通过独特的比较新鲜衍生的 GSC 和正常生发基质祖细胞中的开放染色质，我们能够区分 GSC 中与细胞迁移特别相关的转录可及区域（最终修订版）。这使我们能够推断出肿瘤特异性调控区域内最显着的转录因子（TF）基序与移民有关，TEAD 家族成为首选。 TEAD 特遣队及其合作伙伴激活剂 YAP/TAZ 是 Hippo 通路的主要效应子，并已对其致癌性进行了研究调节作用主要在大脑之外。测试 YAP-TEAD 在 GBM 生长中的功能作用迁移，我们使用 CRISPR/Cas9 在患者来源的 GSC 中产生 TEAD1 敲除，并采用 YAP-TEAD 抑制剂维替泊芬 (Verteporfin) 可抑制 TEAD 活性。使用这两种方法来抑制 TEAD1 活性，我们检测到细胞体外迁移能力下降以及 EGFR 和 pERK 表达的强烈下调。 EGFR 敲除同样表现出体外迁移减少以及 TEAD 活性降低。基于此，我们假设 TEAD 活性调节 GBM 的浸润性生长，介导至少部分通过 EGFR 调节环路。为了检验这个假设，我们的目标是 1）系统地定义 GBM 亚型开放染色质和下游 TEAD 靶基因的 TEAD 占据情况； 2a) 解剖 Hippo-YAP/TAZ-TEAD 与 EGFR/RTK-ERK 之间的机制关系； 2b) 定义角色 TEAD1在体内外调节迁移和生长及其与EGFR的协同作用； 3）测试 YAP-TEAD 抑制剂（例如维替泊芬）的潜在治疗功效。我们设想 YAP/TAZ-TEAD 作为 GBM 浸润性生长的重要调节因子，药物抑制其活性可减轻肿瘤减轻RTK的致癌作用，从而为GBM患者提供新的治疗选择。