Carbonic Anhydrase IX as an Anticancer Therapy Target: Preclinical Evaluation of Internalizing Monoclonal Antibody Directed to Catalytic Domain

M.      Zatovicova; L.      Jelenska; A.      Hulikova; L.      Csaderova; Z.      Ditte; P.      Ditte; T.      Goliasova; J.      Pastorek; S.      Pastorekova

doi:10.2174/138161210793429832

Abstract

Carbonic anhydrase IX (CA IX) is a suitable target for various anticancer strategies. It is a cell surface protein that is present in human tumors, but not in the corresponding normal tissues. Expression of CA IX is induced by hypoxia and correlates with cancer prognosis in many tumor types. Moreover, CA IX is functionally implicated in cancer progression as a pro-survival factor protecting cancer cells against hypoxia and acidosis via its capability to regulate pH and cell adhesion. Cancer-related distribution of CA IX allows for targeting cancer cells by antibodies binding to its extracellular domain, whereas functional involvement of CA IX opens the possibility to hit cancer cells by blocking their adaptation to physiologic stresses via inhibition of CA IX enzyme activity. The latter strategy is recently receiving considerable attention and great efforts are made to produce CA IX-selective inhibitor derivatives with anticancer effects. On the other hand, targeting CA IX-expressing cells by immunotherapy has reached clinical trials and is close to application in treatment of renal cell carcinoma patients. Nevertheless, development and characterization of new CA IX-specific antibodies is still ongoing. Here we describe a mouse monoclonal antibody VII/20 directed to catalytic domain of CA IX. We show that upon binding to CA IX, the VII/20 MAb undergoes efficient receptor-mediated internalization, which is a process regulating abundance and signaling of cell surface proteins and has a considerable impact on immunotherapy. We evaluated biological properties of the MAb and demonstrated its capacity to elicit anti-cancer effect in mouse xenograft model of colorectal carcinoma. Thus, the VII/20 MAb might serve as a tool for preclinical studies of immunotherapeutic strategies against non-RCC tumors. These have not been explored so far and include broad spectrum of cancer types, treatment of which might benefit from CA IX-mediated targeting.

Keywords: Carbonic anhydrase IX, cancer, hypoxia, monoclonal antibody, immunotherapy, anticancer, acidosis, xenograft model, colorectal carcinoma, histidine residues, gastrointestinal tract, epithelia, hypoxia-inducible transcription factor, hypoxic stress, normoxia, metalloproteinase, extracellular domain (ECD), transmembrane region (TM), intracytoplasmic (IC), catalytic domain (CA), proteoglycan aggreccan, keratan sulfate, intrinsic buffer, homologous S-S bridge, tumor microenvironment, hypoxic tumour microenvironment, anaerobic glycolysis, luminal epithelium, Sulfonamides, isoenzymes, topology, celecoxib, valdecoxib, arachidonic acid, Fluorescein-conjugated carbonic anhydrase inhibitor thioureido-homosulfanilamide (FITC-CAI), acetazolamide, human IgG, receptor-mediated internalization, monoclonal antibodies, tumor xenografts, heterogeneity, immunoconjugates