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：
10543769
负责人：
Nadejda Mincheva Tsankova
金额：
$ 37.08万
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 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10543769
关键词：
ATAC-seq Adhesions Affect Attenuated Automobile Driving Binding Biochemical Biology Brain Brain Neoplasms CRISPR/Cas technology Carcinoma Cell Culture Techniques Cells ChIP-seq Chemoresistance Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Complex Data Dependence Development Diffuse Disease Progression Down-Regulation EGF gene Elements Epidermal Growth Factor Receptor Epigenetic Process Excision Extracellular Matrix FDA approved Family Feedback Gene Expression Gene Targeting Genes Genetic Transcription Glioblastoma Glioma Growth Histology Human Hypoxia Immunocompetent In Vitro Infiltration Infiltrative Growth Invaded Knock-in Knock-out Ligands Link Malignant Neoplasms Measures Mediating Mesenchymal Differentiation Metabolic Metabolism Methods Modeling Molecular Morphologic artifacts Mus Nucleic Acid Regulatory Sequences Oncogenes Oncogenic Operative Surgical Procedures PDGFRA gene Pathway interactions Patients Penetrance Proliferating Proto-Oncogene Proteins c-akt Recurrent tumor Role SCID Mice Sampling Signal Transduction Slice Solid Neoplasm System Testing Tissues Toxic effect Treatment Efficacy Tumor Burden Tumor Cell Biology Verteporfin Xenograft procedure cell growth cell motility chemotherapy genome-wide in vivo in vivo engraftment inhibitor insight migration neoplastic cell nerve stem cell neuroinflammation novel novel therapeutics overexpression pharmacologic prevent progenitor programs promoter prospective stem cell population stem cell proliferation stressor synergism tool transcription factor transcriptome sequencing tumor tumor growth tumor progression

项目摘要

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.

胶质母细胞瘤（GBM）肿瘤表现出较大的增殖性核心和一个扩散浸润的边界，后者导致不完整的手术切除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会提供新颖的，并可能提供更多有效的治疗机会。肿瘤细胞浸润/迁移的生物学很复杂且紧密与其微环境有关，包括适应独特的代谢，低氧，运动性，细胞外的基质和神经炎症应激源。在依赖的神经胶质瘤模型中，这些途径很难概括关于细胞培养。随着细胞命运的决定在表观遗传控制的水平上汇聚在一起，一种有吸引力的策略研究复杂的迁移生物学是使用表观遗传学来定义在与天然肿瘤细胞状态相似的系统中迁移。在此提案中，我们利用我们实验室的表观遗传学专业知识，用于研究神经胶质瘤干细胞种群（GSC）的细胞命运状态，直接从其肿瘤生态位分离而没有培养前伪影[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可转录区域（在最终修订中）。这允许我们推断肿瘤特异性调节区域内最显着的转录因子（TF）基序与移民有关，Tead家族成为顶级候选人。 Tead TFS，以及他们的共同激活剂yap/taz，是河马途径的主要效应子，已经对其致癌性进行了研究调节作用主要是在大脑之外。测试Yap-tead在GBM增长中的功能作用和迁移，我们使用CRISPR/CAS9在患者来源的GSC中产生了Tead1敲除，并采用了 yap-tead抑制剂verteporfin，以抑制tead活性。使用这两种方法抑制tead1活性，我们检测到细胞在体外迁移的能力降低，EGFR和PERK表达的稳健下调。 EGFR敲除同样显示出体外迁移的降低，并且tead活性降低。基于这一点，我们假设将活动调节GBM的浸润性增长，并介导通过EGFR调节循环最少部分。为了检验这一假设，我们的目的是1）系统定义跨GBM亚型的开放染色质和下游Tead-tagt靶基因的占用率； 2a）解剖从机械上讲，Hippo-YAP/TAZ-TEAD与EGFR/RTK-ERK之间的关系； 2b）定义 TEAD1在体外和/ex vivo中调节迁移和生长以及与EGFR的协同作用； 3）测试 YAP-TEAD抑制剂（例如Verteporfin）的潜在治疗功效。我们设想yap/taz-tead GBM中浸润性生长的重要调节剂，药理学抑制其活性会减轻肿瘤负担同时也阻止了致癌性RTK效应，从而为GBM患者提供了新的治疗选择。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Chromosomal instability in adult-type diffuse gliomas.

DOI：
10.1186/s40478-022-01420-w
发表时间：
2022-08-17
期刊：
Acta neuropathologica communications
影响因子：
7.1
作者：
通讯作者：

Pathophysiology of SARS-CoV-2: the Mount Sinai COVID-19 autopsy experience.

DOI：
10.1038/s41379-021-00793-y
发表时间：
2021-08
期刊：
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc
影响因子：
0
作者：
Bryce C;Grimes Z;Pujadas E;Ahuja S;Beasley MB;Albrecht R;Hernandez T;Stock A;Zhao Z;AlRasheed MR;Chen J;Li L;Wang D;Corben A;Haines GK 3rd;Westra WH;Umphlett M;Gordon RE;Reidy J;Petersen B;Salem F;Fiel MI;El Jamal SM;Tsankova NM;Houldsworth J;Mussa Z;Veremis B;Sordillo E;Gitman MR;Nowak M;Brody R;Harpaz N;Merad M;Gnjatic S;Liu WC;Schotsaert M;Miorin L;Aydillo Gomez TA;Ramos-Lopez I;Garcia-Sastre A;Donnelly R;Seigler P;Keys C;Cameron J;Moultrie I;Washington KL;Treatman J;Sebra R;Jhang J;Firpo A;Lednicky J;Paniz-Mondolfi A;Cordon-Cardo C;Fowkes ME
通讯作者：
Fowkes ME

IDH-mutant astrocytoma with EGFR amplification-Genomic profiling in four cases and review of literature.

DOI：
10.1093/noajnl/vdac067
发表时间：
2022-01
期刊：
NEURO-ONCOLOGY ADVANCES
影响因子：
0
作者：
Umphlett, Melissa;Bilal, Khawaja Hasan;Martini, Michael L.;Suwala, Abigail K.;Ahuja, Sadhna;Rashidipour, Omid;Germano, Isabelle;Snuderl, Matija;Morgenstern, Peter;Tsankova, Nadejda M.
通讯作者：
Tsankova, Nadejda M.

An atlas of late prenatal human neurodevelopment resolved by single-nucleus transcriptomics.

DOI：
10.1038/s41467-022-34975-2
发表时间：
2022-12-12
期刊：
NATURE COMMUNICATIONS
影响因子：
16.6
作者：
Ramos, Susana, I;Mussa, Zarmeen M.;Falk, Elisa N.;Pai, Balagopal;Giotti, Bruno;Allette, Kimaada;Cai, Peiwen;Dekio, Fumiko;Sebra, Robert;Beaumont, Kristin G.;Tsankov, Alexander M.;Tsankova, Nadejda M.
通讯作者：
Tsankova, Nadejda M.

FACS-based Isolation of Neural and Glioma Stem Cell Populations from Fresh Human Tissues Utilizing EGF Ligand.