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.

项目概要/摘要多形性胶质母细胞瘤 (GBM) 是一种侵袭性、遗传多样性、普遍致命的疾病 GBM 中观察到的侵袭性特征和遗传多样性。与具有干细胞样特征的细胞群（称为胶质母细胞瘤干细胞）有关（GSC）与 Rich 实验室合作，我们证明了 EYA2（眼睛缺失转录）。与分化的胶质母细胞瘤相比，共激活剂和磷酸酶 2) 在 GSC 中高表达细胞（DGC）和神经干细胞（NSC），其高度定位于中心体。 GSC 的增殖特别依赖于 EYA2 的酪氨酸磷酸酶 (Tyr Ptase) 活性，因为 EYA2 Tyr Ptase 活性的药理学抑制或 EYA2 Tyr Ptase dead 的过度表达突变体导致细胞周期停滞、单极和多极纺锤体异常以及细胞凋亡与 DGC 和 NSC 相比，这些数据共同提供了酪氨酸酶活性的证据。 EYA2 在中心体引发 GSC 增殖所需的新功能。我们之前已经证明，几种癌症重新表达 EYA 蛋白以促进 Tyr 通过多种细胞质和核功能产生致瘤和转移表型。 EYA 蛋白的 Ptase 活性与多种癌症的细胞周期进展有关，但有趣的是，这些表型的分子原因大多未知。之前已显示其在中心体发挥作用，我的数据强调 EYA2 定位于中心体。 GSC 中的中心体，并且抑制 EYA2 的酪氨酸酶活性会导致中心体有丝分裂 GSC 中的碎片，表明 EYA2 在癌症中具有新的有丝分裂调节作用。此外，使用 EYA2 底物捕获实验，我已经鉴定了几个中心体相关蛋白和有丝分裂调节因子作为 EYA2 酪氨酸酶活性的潜在底物。因为 EYA2 在中心体和有丝分裂纺锤体中酪氨酸酶活性的精确机制形成仍然难以捉摸，在这个提案中，我将测试 EYA2 的酪氨酸的假设中心体成熟直接需要磷酸酶活性有丝分裂纺锤体的形成，并且 EYA2 Tyr Ptase 活性的直接目标可以是确定对有丝分裂进展至关重要。