Elucidating the role of EYA2 at centrosomes in glioblastoma stem cells

阐明 EYA2 在胶质母细胞瘤干细胞中心体的作用

基本信息

批准号：
10751806
负责人：
Arthur R Wolin
金额：
$ 3.59万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751806
关键词：
2-tyrosine 3&apos Untranslated Regions AKAP9 gene Adult Antibodies Apoptosis Binding Biological Assay Biology Brain Cell Cycle Arrest Cell Cycle Progression Cells Centrosome Characteristics Chromosome Segregation Co-Immunoprecipitations Collaborations Cytoplasm Data Defect Development Disease Exhibits Eye Foundations G2/M Arrest Genetic Variation Glioblastoma In Vitro Link Malignant Neoplasms Mass Spectrum Analysis Mitosis Mitotic Mitotic spindle Molecular Nuclear PLK1 gene Peptides Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphotyrosine Population Proliferating Protein Tyrosine Phosphatase Proteins Recurrent tumor Role Testing Time Transcription Coactivator Trimethoprim-Sulfamethoxazole Western Blotting Work efficacious treatment experimental study immunocytochemistry inhibitor knock-down mimetics mutant nerve stem cell novel overexpression pharmacologic progenitor radioresistant small hairpin RNA stem stem cell proliferation stem cells survival outcome therapeutic target tumorigenic

项目摘要

PROJECT SUMMARY/ABSTRACT Glioblastoma multiforme (GBM) is an aggressive, genetically diverse, and universally lethal malignancy of the brain. The aggressive characteristics and genetic diversity observed in GBM have been linked to a population of cells with stem-like characteristics known as glioblastoma stem cells (GSCs). In collaboration with the Rich lab, we have shown that EYA2 (eyes absent transcriptional coactivator and phosphatase 2) is highly expressed in GSCs compared to differentiated glioblastoma cells (DGCs) and neural stem cells (NSCs), where it is highly localized to centrosomes. Importantly, GSCs are uniquely reliant on EYA2’s tyrosine phosphatase (Tyr Ptase) activity for proliferation, as pharmacological inhibition of EYA2’s Tyr Ptase activity or overexpression of an EYA2 Tyr Ptase dead mutant causes cell cycle arrest, mono- and multipolar spindle abnormalities, and apoptosis when compared to DGCs and NSCs. Together, these data provide evidence that the Tyr Ptase activity of EYA2 elicits a novel function at centrosomes that is required for GSC proliferation. We have previously demonstrated that several cancers re-express EYA proteins to promote tumorigenic and metastatic phenotypes through multiple cytoplasmic and nuclear functions. The Tyr Ptase activity of EYA proteins is implicated in cell cycle progression in several cancers, but the molecular cause of these phenotypes is mostly unknown. Intriguingly, EYA proteins have not previously been shown to function at centrosomes. My data highlights that EYA2 is localized to centrosomes in GSCs and that inhibition of EYA2’s Tyr Ptase activity leads to centrosome fragmentation in mitotic GSCs, suggesting a novel, mitotic regulatory role for EYA2 in cancer. Furthermore, using EYA2 substrate-trapping experiments I have identified several centrosome- associated proteins and mitotic regulators as potential substrates of EYA2’s Tyr Ptase activity. Because the precise mechanism for EYA2’s Tyr Ptase activity at centrosomes and in mitotic spindle formation remains elusive, in this proposal I will test the hypothesis that EYA2’s tyrosine phosphatase activity is directly required at centrosomes for centrosome maturation and mitotic spindle formation, and that direct targets of the EYA2 Tyr Ptase activity can be identified that are critical for mitotic progression.

项目摘要/摘要胶质母细胞瘤多形（GBM）是一种侵略性，遗传多样性，普遍致命的大脑的恶性肿瘤。在GBM中观察到的积极特征和遗传多样性具有与具有胶质母细胞瘤干细胞的干细胞群有关（GSC）。与富实验室合作，我们表明了EYA2（眼睛没有转录与分化的胶质母细胞瘤相比细胞（DGC）和神经元干细胞（NSC），在该干细胞中高度局限于中心体。重要的是， GSC独特地依赖于Eya2的酪氨酸磷酸酶（Tyr PTase）活性，如 EYA2的Tyr PTase活性或EYA2 Tyr PTase死亡的药理抑制作用突变体引起细胞周期停滞，单极纺锤体异常和凋亡与DGC和NSC相比。这些数据共同提供了证据表明 EYA2引起了GSC增殖所需的中心体的新功能。我们以前已经证明了几种癌症重新表达EYA蛋白以促进通过多种细胞质和核功能的致瘤和转移表型。 tyr EYA蛋白的PTase活性在几种癌症的细胞周期进程中暗示，但是这些表型的分子原因大多是未知的。有趣的是，Eya蛋白没有以前已显示在中心体处发挥作用。我的数据强调了Eya2本地化 GSC中的中心体和抑制Eya2的Tyr PTase活性会导致中心体有丝分裂GSC的碎片，表明EYA2在癌症中的新型有丝分裂调节作用。此外，使用EYA2底物捕捞实验，我已经确定了几个中心体 - 相关的蛋白质和有丝分裂调节剂是EYA2的Tyr PTase活性的潜在底物。因为Eya2在中心体和有丝分裂纺锤体中的Tyr PTase活性的精确机制形成仍然难以捉摸，在此提案中，我将检验以下假设：Eya2的酪氨酸磷酸酶活性直接需要在中心体中进行中心体成熟和有丝分裂主轴形成，以及Eya2 Tyr PTase活性的直接靶标可以是确定对有丝分裂进展至关重要。