NEDD9 Depletion Leads to MMP14 Inactivation by TIMP2 and Prevents Invasion and Metastasis

Abstract

The scaffolding protein NEDD9 is an established pro-metastatic marker in several cancers. Nevertheless, the molecular mechanisms of NEDD9 driven metastasis in cancers remain ill defined. Here, using a comprehensive breast cancer (BCa) tissue microarray, it was show that increased levels of NEDD9 protein significantly correlated with the transition from carcinoma in situ to invasive carcinoma. Similarly, it was shown that NEDD9 overexpression is a hallmark of highly invasive BCa cells. Moreover, NEDD9 expression is crucial for the protease-dependent mesenchymal invasion of cancer cells at the primary site but not at the metastatic site. Depletion of NEDD9 is sufficient to suppress invasion of tumor cells in vitro and in vivo, leading to decreased circulating tumor cells (CTCs) and lung metastases in xenograft models. Mechanistically, NEDD9 localized to invasive pseudopods and was required for local matrix degradation. Depletion of NEDD9 impaired invasion of cancer cells through inactivation of membrane-bound matrix metalloproteinase MMP14 by excess TIMP2 on the cell surface. Inactivation of MMP14 is accompanied by reduced collagenolytic activity of soluble metalloproteinases MMP2 and MMP9. Re-expression of NEDD9 is sufficient to restore the activity of MMP14 and the invasive properties of BCa cells in vitro and in vivo. Collectively, these findings uncover critical steps in NEDD9-dependent invasion of BCa cells. Implications This study provides a mechanistic basis for potential therapeutic interventions to prevent metastasis.

Prometastatic NEDD9 Regulates Individual Cell Migration via Caveolin-1–Dependent Trafficking of Integrins

“…Overexpression of NEDD9 in MDA-MB-453 and ZR75-1 BC cells with low endogenous expression of NEDD9 using a doxycycline inducible system led to a significant increase in migration (Fig.1K–L). In addition to the decrease in 2D migration, NEDD9-depleted cells exhibited up to 60% decrease in 3D migration/invasion through matrigel (Fig.S1C), confirming our previous observations [7]. The decrease in cell migration could be due to the impairment in cell adhesion to certain components of the ECM.…”

“…Overexpression of the NEDD9 protein is known to promote migration/invasion in multiple cancers [5, 7, 8]. We have recently reported that depletion of NEDD9 leads to a significant decrease in breast cancer metastasis due to inactivation of MMPs [7] and migration deficiency, but the molecular mechanisms of the phenomenon are not completely understood.…”

“…We have recently reported that depletion of NEDD9 leads to a significant decrease in breast cancer metastasis due to inactivation of MMPs [7] and migration deficiency, but the molecular mechanisms of the phenomenon are not completely understood. The NEDD9-dependent migration was studied downstream of NEDD9 and its role in the upstream integrin signaling was unclear, particularly for breast cancers.…”

“…Upregulation of NEDD9 promotes epithelial to mesenchymal transition, migration, and metastasis in melanoma, glioblastoma, colon cancer and head and neck squamous carcinoma [3, 4] [5–7]. Genetic ablation of Nedd9 in a Her2-driven spontaneous mammary tumor model decreases mammary tumor incidence, initiation rate and tumor burden [8], indicating a tumor promoting role of NEDD9.…”

Digging a little deeper: The stages of invadopodium formation and maturation

“…ILK recruits the scaffold protein IQGAP to invadopodia to induce membrane type-I MMP (MT1-MMP) exocytosis, while NEDD9 limits the accumulation of tissue inhibitor of metalloproteinase-2 (TIMP2), an endogenous MT1-MMP inhibitor, at invadopodia to promote MT1-MMP-mediated matrix degradation (Branch et al, 2012; McLaughlin et al, 2014). Taken together, adhesion proteins regulate invadopodium maturation by enhancing actin polymerization and MMP-mediated matrix degradation, while having limited effects on invadopodium precursor assembly.…”