Three-Dimensional Cell Culture-Based Screening Identifies the Anthelmintic Drug Nitazoxanide as a Candidate for Treatment of Colorectal Cancer

• Published in: Molecular cancer therapeutics 2015-06, Vol.14 (6), p.1504-1516
• Main Authors: Senkowski, Wojciech, Zhang, Xiaonan, Olofsson, Maria Hägg, Isacson, Ruben, Höglund, Urban, Gustafsson, Mats, Nygren, Peter, Linder, Stig, Larsson, Rolf, Fryknäs, Mårten
• Format: Article
• Language: English
• Subjects: Administration, Oral; Animals; Anthelmintics - administration & dosage; Anthelmintics - pharmacokinetics; Anthelmintics - pharmacology; Antineoplastic Agents - administration & dosage; Antineoplastic Agents - pharmacokinetics; Antineoplastic Agents - pharmacology; Cell Culture Techniques - methods; Cell Hypoxia; Cell Survival - drug effects; Colorectal Neoplasms - drug therapy; Colorectal Neoplasms - genetics; Colorectal Neoplasms - pathology; Dose-Response Relationship, Drug; Drug Repositioning - methods; Drug Screening Assays, Antitumor - methods; Female; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; HCT116 Cells; HT29 Cells; Humans; Mice, Inbred Strains; Mice, Nude; Microscopy, Fluorescence; Oxidative Phosphorylation - drug effects; Signal Transduction - drug effects; Spheroids, Cellular - drug effects; Spheroids, Cellular - metabolism; Thiazoles - administration & dosage; Thiazoles - pharmacokinetics; Thiazoles - pharmacology; Xenograft Model Antitumor Assays
• ISSN: 1535-7163; 1538-8514; 1538-8514
• DOI: 10.1158/1535-7163.MCT-14-0792

Because dormant cancer cells in hypoxic and nutrient-deprived regions of solid tumors provide a major obstacle to treatment, compounds targeting those cells might have clinical benefits. Here, we describe a high-throughput drug screening approach, using glucose-deprived multicellular tumor spheroids (MCTS) with inner hypoxia, to identify compounds that specifically target this cell population. We used a concept of drug repositioning-using known molecules for new indications. This is a promising strategy to identify molecules for rapid clinical advancement. By screening 1,600 compounds with documented clinical history, we aimed to identify candidates with unforeseen potential for repositioning as anticancer drugs. Our screen identified five molecules with pronounced MCTS-selective activity: nitazoxanide, niclosamide, closantel, pyrvinium pamoate, and salinomycin. Herein, we show that all five compounds inhibit mitochondrial respiration. This suggests that cancer cells in low glucose concentrations depend on oxidative phosphorylation rather than solely glycolysis. Importantly, continuous exposure to the compounds was required to achieve effective treatment. Nitazoxanide, an FDA-approved antiprotozoal drug with excellent pharmacokinetic and safety profile, is the only molecule among the screening hits that reaches high plasma concentrations persisting for up to a few hours after single oral dose. Nitazoxanide activated the AMPK pathway and downregulated c-Myc, mTOR, and Wnt signaling at clinically achievable concentrations. Nitazoxanide combined with the cytotoxic drug irinotecan showed anticancer activity in vivo. We here report that the FDA-approved anthelmintic drug nitazoxanide could be a potential candidate for advancement into cancer clinical trials.