Inhibition of carbonic anhydrases IX/XII by SLC-0111 boosts cisplatin effects in hampering head and neck squamous carcinoma cell growth and invasion

Abstract

Background Hypoxic tumor microenvironment (TME) contributes to the onset of many aspects of the cancer biology associated to the resistance to conventional therapies. Hypoxia is a common characteristic and negative prognostic factor in the head and neck squamous carcinomas (HNSCC) and is correlated with aggressive and invasive phenotype as well as with failure to chemo- and radio-therapies. The carbonic anhydrase isoenzymes IX and XII (CA IX/XII), regulators of extra and intracellular pH, are overexpressed in TME and are involved in adaptative changes occurring in cancer cells to survive at low O2. In this study, we aim to investigate in HNSCC cells and murine models the possibility to target CA IX/XII by the specific inhibitor SLC-0111 to potentiate the effects of cisplatin in hampering cell growth, migration and invasion. Furthermore, we analyzed the signal pathways cooperating in acquisition of a more aggressive phenotype including stemness, epithelial-mesenchymal transition and apoptotic markers. Methods The effects of cisplatin, CA IX/XII specific inhibitor SLC-0111, and the combinatorial treatment were tested on proliferation, migration, invasion of HNSCC cells grown in 2D and 3D models. Main signal pathways and the expression of stemness, mesenchymal and apoptotic markers were analyzed by western blotting. Molecular imaging using NIR-Annexin V and NIR-Prosense was performed in HNSCC xenografts to detect tumor growth and metastatic spread. Results HNSCC cells grown in 2D and 3D models under hypoxic conditions showed increased levels of CA IX/XII and greater resistance to cisplatin than cells grown under normoxic conditions. The addition of CA IX/XII inhibitor SLC-0111 to cisplatin sensitized HNSCC cells to the chemotherapeutic agent and caused a reduction of proliferation, migration and invasiveness. Furthermore, the combination therapy hampered activation of STAT3, AKT, ERK, and EMT program, whereas it induced apoptosis. In HNSCC xenografts the treatment with cisplatin plus SLC-0111 caused an inhibition of tumor growth and an induction of apoptosis as well as a reduction of metastatic spread at a higher extent than single agents. Conclusion Our results highlight the ability of SLC-0111 to sensitize HNSCC to cisplatin by hindering hypoxia-induced signaling network that are shared among mechanisms involved in therapy resistance and metastasis.

Benzenesulfonamides with different rigidity-conferring linkers as carbonic anhydrase inhibitors: an insight into the antiproliferative effect on glioblastoma, pancreatic, and breast cancer cells

“…Enzyme inhibition results confirmed that chemical manipulation of the linker and the tail could modulate the CA activity, despite the presence of a pan-CA inhibiting zinc binding group (sulphonamide). Cell-based data confirmed the relevant antiproliferative properties of this class of CAIs, the importance of the tumour-associated CAs for tumour growth and development, and their inhibition as a fruitful strategy to hamper these diseases, reinforcing previous data 37 , 38 on the significant antitumor effects of this class of CA inhibitors.…”

Cancer Therapeutic Targeting of Hypoxia Induced Carbonic Anhydrase IX: From Bench to Bedside

“…This has been borne out in a number of in vitro and in vivo tumor models. Combining CAIX inhibitors with standard of care chemotherapy agents has been demonstrated to be significantly more efficacious than single agents alone [ 38 , 39 , 40 ]. Thus, combining the CAIX inhibitor, SLC-0111 with paclitaxel or with sunitinib in breast cancer [ 19 , 41 ], with temozolomide in pediatric and adult glioblastoma [ 42 ], with gemicitabine in pancreatic cancer [ 43 ], or an analog of SLC-0111, FC531, with cytarabine or Quizartinib in acute myeloblastic leukemia (AML) [ 44 ], results in greater inhibition of tumor growth and in significant increases in the survival of the tumor bearing mice.…”

Design and synthesis of benzothiazole-based SLC-0111 analogues as new inhibitors for the cancer-associated carbonic anhydrase isoforms IX and XII

“…The ureido benzenesulfonamide SLC-0111 ( Figure 1 ) has been developed utilising the tail approach as the first-in-class h CA IX inhibitor which, for the management of metastatic hypoxic solid tumours, is currently being investigated within phase I/II clinical trials 3 , 23 , 24 . To date, numerous SLC-0111 analogues have been described with the prime aim to enhance potency and selectivity towards h CA IX exploiting bioisosteric replacement approach via replacement of the SLC-0111aryl tail by various sets of chemical scaffolds ( Figure 1 ) like benzofuran I 23 , thiazolo[3,2- a ]benzimidazole II 25 , triazine III 26 , thiazole IV , and thiadiazole V 27 .…”