Impact of hypoxia on chemoresistance of mesothelioma mediated by the proton-coupled folate transporter, and preclinical activity of new anti-LDH-A compounds

• Published in: British journal of cancer Vol. 123; no. 4; pp. 644 - 656
• Main Authors: Li Petri, Giovanna, El Hassouni, Btissame, Sciarrillo, Rocco, Funel, Niccola, Mantini, Giulia, Zeeuw van der Laan, Eveline A., Cascioferro, Stella, Avan, Amir, Zucali, Paolo Andrea, Zaffaroni, Nadia, Lagerweij, Tonny, Parrino, Barbara, Smid, Kees, Deraco, Marcello, Granchi, Carlotta, Braczko, Alicja, Smolenski, Ryszard T., Matherly, Larry H., Jansen, Gerrit, Assaraf, Yehuda G., Diana, Patrizia, Cloos, Jacqueline, Peters, Godefridus J., Minutolo, Filippo, Giovannetti, Elisa
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 18.08.2020; Nature Publishing Group
• Subjects: 631/67/1641; 692/308/575; Animal models; Animals; Antigens, Neoplasm - genetics; Antigens, Neoplasm - metabolism; Biomedical and Life Sciences; Biomedicine; Cancer Research; Carbonic Anhydrase IX - genetics; Carbonic Anhydrase IX - metabolism; Cell Culture Techniques; Cell Hypoxia; Cell Line, Tumor; Chemoresistance; Deoxycytidine - administration & dosage; Deoxycytidine - analogs & derivatives; Deoxycytidine - pharmacology; DNA microarrays; Drug Resistance; Drug Resistance, Neoplasm - drug effects; Drug Synergism; Enzyme Inhibitors - administration & dosage; Enzyme Inhibitors - pharmacology; Epidemiology; Female; Folic acid; Gastric cancer; Gemcitabine; Gene expression; Gene Expression Regulation, Neoplastic - drug effects; Glycolysis; Humans; Hypoxia; L-Lactate dehydrogenase; L-Lactate Dehydrogenase - genetics; Lactic acid; Mesothelioma; Mesothelioma, Malignant - drug therapy; Mesothelioma, Malignant - genetics; Mesothelioma, Malignant - metabolism; Mice; Molecular Medicine; Oncology; Pemetrexed - administration & dosage; Pemetrexed - pharmacology; Peritoneal Neoplasms - drug therapy; Peritoneal Neoplasms - genetics; Peritoneal Neoplasms - metabolism; Peritoneum; Pleural Neoplasms - drug therapy; Pleural Neoplasms - genetics; Pleural Neoplasms - metabolism; Proton-Coupled Folate Transporter - genetics; Proton-Coupled Folate Transporter - metabolism; Spheroids; Tumors; Xenograft Model Antitumor Assays
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/s41416-020-0912-9

Background Expression of proton-coupled folate transporter (PCFT) is associated with survival of mesothelioma patients treated with pemetrexed, and is reduced by hypoxia, prompting studies to elucidate their correlation. Methods Modulation of glycolytic gene expression was evaluated by PCR arrays in tumour cells and primary cultures growing under hypoxia, in spheroids and after PCFT silencing. Inhibitors of lactate dehydrogenase (LDH-A) were tested in vitro and in vivo. LDH-A expression was determined in tissue microarrays of radically resected malignant pleural mesothelioma (MPM, N  = 33) and diffuse peritoneal mesothelioma (DMPM, N  = 56) patients. Results Overexpression of hypoxia marker CAIX was associated with low PCFT expression and decreased MPM cell growth inhibition by pemetrexed. Through integration of PCR arrays in hypoxic cells and spheroids and following PCFT silencing, we identified the upregulation of LDH-A, which correlated with shorter survival of MPM and DMPM patients. Novel LDH-A inhibitors enhanced spheroid disintegration and displayed synergistic effects with pemetrexed in MPM and gemcitabine in DMPM cells. Studies with bioluminescent hypoxic orthotopic and subcutaneous DMPM athymic-mice models revealed the marked antitumour activity of the LDH-A inhibitor NHI-Glc-2, alone or combined with gemcitabine. Conclusions This study provides novel insights into hypoxia/PCFT-dependent chemoresistance, unravelling the potential prognostic value of LDH-A, and demonstrating the preclinical activity of LDH-A inhibitors.