CLIC3 controls recycling of late endosomal MT1-MMP and dictates invasion and metastasis in breast cancer

Abstract

Chloride intracellular channel 3 (CLIC3) drives invasiveness of pancreatic and ovarian cancer by acting in concert with Rab25 to regulate the recycling of a5b1 integrin from late endosomes to the plasma membrane. Here, we show that in two estrogen receptor (ER)-negative breast cancer cell lines, CLIC3 has little influence on integrin recycling, but controls trafficking of the pro-invasive matrix metalloproteinase MT1-MMP (also known as MMP14). In MDA-MB-231 cells, MT1-MMP and CLIC3 are localized primarily to late endosomal/lysosomal compartments located above the plane of adhesion and near the nucleus. MT1-MMP is transferred from these late endosomes to sites of cell-matrix adhesion in a CLIC3-dependent fashion. Correspondingly, CLIC3-knockdown opposes MT1-MMP-dependent invasive processes. These include the disruption of the basement membrane as acini formed from MCF10DCIS.com cells acquire invasive characteristics in 3D culture, and the invasion of MDA-MB-231 cells into Matrigel or organotypic plugs of type I collagen. Consistent with this, expression of CLIC3 predicts poor prognosis in ER-negative breast cancer. The identification of MT1-MMP as a cargo of a CLIC3-regulated pathway that drives invasion highlights the importance of late endosomal sorting and trafficking in breast cancer.

Mechanisms and functions of lysosome positioning

“…Secreted acid hydrolases are thought to degrade the extracellular matrix, facilitating cell migration and invasion (Mohamed and Sloane, 2006;Dykes et al, 2016). Transport of the transmembrane type 1 matrix metalloproteinase (MT1-MMP, also known as MMP14) from late endosomes to plasma membrane invadopodia also contributes to cancer cell invasiveness and metastasis (Monteiro et al, 2013;Macpherson et al, 2014). Furthermore, some integrins that regulate cell adhesion and migration by attachment to the extracellular matrix traffic through late endosomes and lysosomes (Dozynkiewicz et al, 2012), possibly depending on lysosome exocytosis for their delivery to the plasma membrane.…”

Microtubules in 3D cell motility

“…Moreover, MT1-MMP in conjunction with the physical properties of ECM is a determining factor in mesenchymal cell migration in 3D (Wolf et al, 2013). MT1-MMP trafficking and delivery to the plasma membrane are microtubule dependent, involve late endosomes and are required for mesenchymal invasion by cancer cells (Frittoli et al, 2014;Macpherson et al, 2014;Remacle et al, 2005;Rosse et al, 2014). It should be noted that delivery of MT1-MMP to the plasma membrane is also controlled by F-actin and its regulator N-WASP (Yu et al, 2012), and the respective contributions of microtubule-and actin-based mechanisms of MT1-MMP trafficking in 3D will thus have to be further investigated.…”

Role of cholesterol in SNARE-mediated trafficking on intracellular membranes

“…Chavrier and co-workers have established that VAMP7 and the exocyst complex deliver a late endosomal pool of membrane type 1 metalloproteinase (MT1-MMP, also known as MMP14) to the cell surface for ECM proteolysis, which is crucial for invasive migration (Steffen et al, 2008). Earlier steps in this pathway might involve CLIC3 (Macpherson et al, 2014). Later trafficking events probably require the actin cytoskeleton, as following its transit from Rab7-positive endosomes to the plasma membrane, MT1-MMP is retained in invasive protrusions by direct tethering to F-actin (Yu et al, 2012).…”