CRN2 binds to TIMP4 and MMP14 and promotes perivascular invasion of glioblastoma cells.
CRN2 binds to TIMP4 and MMP14 and promotes perivascular invasion of glioblastoma cells.
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DOI：
10.1016/j.ejcb.2019.151046
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发表时间：
2019-10
影响因子：
6.6
通讯作者：
Roxana Solga;J. Behrens;Anja Ziemann;A. Riou;C. Berwanger;L. Becker;L. Garrett;M. D. de Angelis;L. Fischer;R. Coras;Katalin Barkovits;K. Marcus;E. Mahabir;L. Eichinger;R. Schröder;A. Noegel;C. Clemen
中科院分区：
文献类型：
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作者：
Roxana Solga;J. Behrens;Anja Ziemann;A. Riou;C. Berwanger;L. Becker;L. Garrett;M. D. de Angelis;L. Fischer;R. Coras;Katalin Barkovits;K. Marcus;E. Mahabir;L. Eichinger;R. Schröder;A. Noegel;C. Clemen
CRN2 是一种肌动蛋白丝结合蛋白，参与多种细胞过程的调节，包括细胞迁移和侵袭。 CRN2 与不同类型人类癌症的恶性进展有关。我们使用 CRN2 敲除小鼠进行分析，并与 Tp53/Pten 敲除胶质母细胞瘤小鼠模型进行杂交。 CRN2 敲除小鼠在德国小鼠诊所进行了表型筛选。通过免疫组织化学、免疫荧光和细胞生物学实验对小鼠胶质母细胞瘤组织标本以及培养的小鼠脑切片和胶质母细胞瘤细胞系进行研究。通过免疫沉淀、pull-down 和酶活性测定研究蛋白质相互作用。 CRN2 敲除小鼠表现出神经和行为改变，例如听力敏感性降低、听觉惊吓反应降低、活动减退和排尿次数减少。虽然有或没有额外的 CRN2 敲除等位基因的胶质母细胞瘤小鼠的存活率没有显着差异，但移植的胶质母细胞瘤细胞中 CRN2 水平的增加导致小鼠脑切片毛细血管中肿瘤细胞的包裹量更高。我们确定了肿瘤微环境的两个重要因素，即基质金属蛋白酶 4 (TIMP4) 的组织抑制剂和基质金属蛋白酶 14 (MMP14，同义词：MT1-MMP)，作为 CRN2 的新型结合伴侣。所有三种蛋白质相互作用并共定位于板状伪足的前端，并且在外泌体中新检测到了 CRN2。在功能水平上，我们证明CRN2增加了TIMP4的分泌以及MMP14的催化活性。我们的结果表明，CRN2 代表多形性胶质母细胞瘤肿瘤细胞微环境中的促侵袭效应器。
CRN2 is an actin filament binding protein involved in the regulation of various cellular processes including cell migration and invasion. CRN2 has been implicated in the malignant progression of different types of human cancer. We used CRN2 knock-out mice for analyses as well as for crossbreeding with a Tp53/Pten knock-out glioblastoma mouse model. CRN2 knock-out mice were subjected to a phenotyping screen at the German Mouse Clinic. Murine glioblastoma tissue specimens as well as cultured murine brain slices and glioblastoma cell lines were investigated by immunohistochemistry, immunofluorescence, and cell biological experiments. Protein interactions were studied by immunoprecipitation, pull-down, and enzyme activity assays. CRN2 knock-out mice displayed neurological and behavioural alterations, e.g. reduced hearing sensitivity, reduced acoustic startle response, hypoactivity, and less frequent urination. While glioblastoma mice with or without the additional CRN2 knock-out allele exhibited no significant difference in their survival rates, the increased levels of CRN2 in transplanted glioblastoma cells caused a higher tumour cell encasement of murine brain slice capillaries. We identified two important factors of the tumour microenvironment, the tissue inhibitor of matrix metalloproteinase 4 (TIMP4) and the matrix metalloproteinase 14 (MMP14, synonym: MT1-MMP), as novel binding partners of CRN2. All three proteins mutually interacted and co-localised at the front of lamellipodia, and CRN2 was newly detected in exosomes. On the functional level, we demonstrate that CRN2 increased the secretion of TIMP4 as well as the catalytic activity of MMP14. Our results imply that CRN2 represents a pro-invasive effector within the tumour cell microenvironment of glioblastoma multiforme.