CRN2 binds to TIMP4 and MMP14 and promotes perivascular invasion of glioblastoma cells

Abstract

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.

Comprehensive analysis of expression, prognosis and immune infiltration for TIMPs in glioblastoma

“…Regarding the relationship between TIMP4 and glioma progression, the directivity of the available findings is inconsistent. TIMP4 binds CRN2 (an actin filament binding protein) and promotes perivascular invasion of GBM cells [ 84 ]. High TIMP4/CD63 coexpression contributes to the poor prognosis of GBM patients [ 85 ].…”

Tumor-Associated Microglia and Macrophages in the Glioblastoma Microenvironment and Their Implications for Therapy

“…Similarly, MMP2 can also be expressed by microglia themselves [106]. Moreover, M2-like macrophages express MMP9 and MMP14, which are also associated with ECM degradation and facilitate cancer cell migration through the TME [107][108][109][110]. Microglia also secrete TGF-β, which has been shown to have an invasion-promoting role in glioblastoma cells both in vitro and in vivo in mice [111].…”

Coronin 1C inhibits melanoma metastasis through regulation of MT1-MMP-containing extracellular vesicle secretion

“…Additionally, Coronin 1C was shown to enhance MT1-MMP trafficking to the cell surface in an orthotopic injection model of breast cancer 87 , which is again seemingly at odds with our results. However, recent data using genetic deletion of Coronin 1C in a GEM model of glioblastoma showed no change in time of survival with or without Coronin 1C 88 , further supporting the idea that Coronin 1C impacts tumor progression differently between cancer types. We postulate that the unique metastatic nature of melanoma and the underlying biology of melanocytes may help explain the context-dependent nature of Coronin 1C function in disease progression and metastasis.…”

“…It is possible that Coronin 1C's influence on the actin cytoskeleton at the PM contributes to the increased exosome secretion in our cells either through interactions with Rab27a, cortactin, and/or F-actin itself. Coronin 1C has even been shown to bind to MT1-MMP directly in glioblastoma cells 88 , but the function of this interaction has not been defined. Future studies will be directed at dissecting the precise mechanisms involved in this process.…”