WDFY2 restrains matrix metalloproteinase secretion and cell invasion by controlling VAMP3-dependent recycling

Marte Sneeggen,Coen Campsteijn,Nina Marie Pedersen,Ellen Margrethe Haugsten,Kay Oliver Schink,Harald Stenmark

doi:10.1038/s41467-019-10794-w

Marte Sneeggen, Coen Campsteijn + Show 4 more

Open Access

https://doi.org/10.1038/s41467-019-10794-w

Copy DOI

Journal: Nature Communications	Publication Date: Jun 28, 2019
Citations: 30	License type: open-access

Affiliation: Oslo University Hospital, University of Oslo

Abstract

Cancer cells secrete matrix metalloproteinases to remodel the extracellular matrix, which enables them to overcome tissue barriers and form metastases. The membrane-bound matrix metalloproteinase MT1-MMP (MMP14) is internalized by endocytosis and recycled in endosomal compartments. It is largely unknown how endosomal sorting and recycling of MT1-MMP are controlled. Here, we show that the endosomal protein WDFY2 controls the recycling of MT1-MMP. WDFY2 localizes to endosomal tubules by binding to membranes enriched in phosphatidylinositol 3-phosphate (PtdIns3P). We identify the v-SNARE VAMP3 as an interaction partner of WDFY2. WDFY2 knockout causes a strong redistribution of VAMP3 into small vesicles near the plasma membrane. This is accompanied by increased, VAMP3-dependent secretion of MT1-MMP, enhanced degradation of extracellular matrix, and increased cell invasion. WDFY2 is frequently lost in metastatic cancers, most predominantly in ovarian and prostate cancer. We propose that WDFY2 acts as a tumor suppressor by serving as a gatekeeper for VAMP3 recycling.

Highlights

Cancer cells secrete matrix metalloproteinases to remodel the extracellular matrix, which enables them to overcome tissue barriers and form metastases
To define the localization of WDFY2 in the endocytic pathway, we transiently transfected hTERT-RPE1 cells with green flourescent protein (GFP)-WDFY2 and performed structured illumination microscopy (SIM) together with APPL1 and EEA1 visualized with antibodies (Fig. 1a)
We provide evidence that tumor suppressors can act by restricting endocytic recycling of Matrix metalloproteinases (MMPs) and thereby preventing cell invasion

Summary

Introduction

Cancer cells secrete matrix metalloproteinases to remodel the extracellular matrix, which enables them to overcome tissue barriers and form metastases. WDFY2 knockout causes a strong redistribution of VAMP3 into small vesicles near the plasma membrane This is accompanied by increased, VAMP3-dependent secretion of MT1-MMP, enhanced degradation of extracellular matrix, and increased cell invasion. We have studied the function of WDFY2, a cytosolic protein that has been described to reside on endocytic vesicles close to the plasma membrane[6] It contains a lipid-binding FYVE domain and seven WD40 repeats which can form a β-propeller. WDFY2 localizes to actin-stabilized endosome tubules positive for the small GTPase RAB4 and shows a preference for highly curved membranes enriched for the lipid phosphatidylinositol 3phosphate (PtdIns3P). It interacts with VAMP3, which directs secretion of endosome-derived cargos, including MT1MMP. We show that loss of WDFY2 leads to enhanced secretion of MT1-MMP and allows cells to actively invade into ECM

Methods

Results

Conclusion