The interactome of metabolic enzyme carbonic anhydrase IX reveals novel roles in tumor cell migration and invadopodia/MMP14-mediated invasion

Abstract

Carbonic anhydrase IX (CAIX) is a hypoxia inducible factor 1-induced, cell surface pH regulating enzyme with an established role in tumor progression and clinical outcome. However, the molecular basis of CAIX-mediated tumor progression remains unclear. Here, we have utilized proximity dependent biotinylation (BioID) to map the CAIX ‘interactome’ in breast cancer cells in order to identify physiologically relevant CAIX-associating proteins with potential roles in tumor progression. High confidence proteins identified include metabolic transporters, β1 integrins, integrin-associated protein CD98hc and matrix metalloprotease 14 (MMP14). Biochemical studies validate the association of CAIX with α2β1 integrin, CD98hc and MMP14, and immunofluorescence microscopy demonstrates colocalization of CAIX with α2β1 integrin and MMP14 in F-actin/cofilin-positive lamellipodia/pseudopodia, and with MMP14 to cortactin/Tks5-positive invadopodia. Modulation of CAIX expression and activity results in significant changes in cell migration, collagen degradation and invasion. Mechanistically, we demonstrate that CAIX associates with MMP14 through potential phosphorylation residues within its intracellular domain, and that CAIX enhances MMP14-mediated collagen degradation by directly contributing hydrogen ions required for MMP14 catalytic activity. These findings establish hypoxia-induced CAIX as a novel metabolic component of cellular migration and invasion structures, and provide new mechanistic insights into its role in tumor cell biology.

TMZ regulates GBM stemness via MMP14‐DLL4‐Notch3 pathway

“…Second, we observed a MMP14‐dependent release of a battery of cellular proteins, some, such as UPAR, CA9 and DKK‐3, have been described in context of MMP14; others, such as IGFBP1, IL1 F7, Prostasin, FUCA1, S100A13 or PlexinB3, have not yet been shown to be linked to MMP14 production. We focused our study on DLL4 as most abundant protein detected in ELISA‐based array and because of strong direct impact of DLL4 on survival of GBM patients .…”

Carbonic Anhydrase IX—Mouse versus Human

“…Numerous studies have confirmed the prominent role of human CA IX in pH regulation, which promotes tumor cell survival [4,5,39]. Moreover, recent studies have also shown the significance of CA IX expression for tumor cell migration and invasion [6,7,40,41]. To validate the existence of the same protein characteristics, we performed a functional analysis of mCA IX.…”

Inhibition of carbonic anhydrase IX targets primary tumors, metastases, and cancer stem cells: Three for the price of one

“…95 The association with 2 1 integrin, CD98hc, and MMP14 was also validated by biochemical studies. More interestingly, this study supported the idea that the observed association and colocalization of CA IX and MMP14 within mature invadopodia enhance MMP14-dependent matrix degradation, invasion, and metastasis, 95 thus adding another piece to the complex puzzle of the CA IX role in tumor biology.…”

“…Additional data on the identification of CA IX-interacting partners were recently published by Swayampakula and co-workers. 95 In this study, by utilizing an unbiased proximity ligation approach, authors found that in breast cancer cells hCA IX associates with several metabolic cell surface transporters, 1 integrins, integrin-associated protein CD98hc, and matrix metalloprotease 14 (MMP14). 95 The association with 2 1 integrin, CD98hc, and MMP14 was also validated by biochemical studies.…”

“…95 In this study, by utilizing an unbiased proximity ligation approach, authors found that in breast cancer cells hCA IX associates with several metabolic cell surface transporters, 1 integrins, integrin-associated protein CD98hc, and matrix metalloprotease 14 (MMP14). 95 The association with 2 1 integrin, CD98hc, and MMP14 was also validated by biochemical studies. More interestingly, this study supported the idea that the observed association and colocalization of CA IX and MMP14 within mature invadopodia enhance MMP14-dependent matrix degradation, invasion, and metastasis, 95 thus adding another piece to the complex puzzle of the CA IX role in tumor biology.…”