Kinomic profile in patient-derived glioma cells during hypoxia reveals c-MET-PI3K dependency for adaptation

Abstract

Hypoxic microenvironment is a hallmark of solid tumors, especially glioblastoma. The strong reliance of glioma-propagating cells (GPCs) on hypoxia-induced survival advantages is potentially exploitable for drug development. Methods: To identify key signaling pathways for hypoxia adaptation by patient-derived GPCs, we performed a kinase inhibitor profiling by screening 188 small molecule inhibitors against 130 different kinases in normoxia and hypoxia. Potential kinase candidates were prioritized for in vitro and in vivo investigations using a ranking algorithm that integrated information from the kinome connectivity network and estimated patients' survival based on expression status. Results: Hypoxic drug screen highlighted extensive modifications of kinomic landscape and a crucial functionality of c-MET-PI3K. c-MET inhibitors diminished phosphorylation of c-MET and PI3K in GPCs subjected to hypoxia, suggesting its role in the hypoxic adaptation of GPCs. Mechanistically, the inhibition of c-MET and PI3K impaired antioxidant defense, leading to oxidative catastrophe and apoptosis. Repurposed c-MET inhibitors PF04217903 and tivantinib exhibited hypoxic-dependent drug synergism with temozolomide, resulting in reduced tumor load and growth of GPC xenografts. Detailed analysis of bulk and single-cell glioblastoma transcriptomes associates the cellular subpopulation over-expressing c-MET with inflamed, hypoxic, metastatic, and stem-like phenotypes. Conclusions: Thus, our “bench to bedside (the use of patient-derived GPCs and xenografts for basic research) and back (validation with independent glioblastoma transcriptome databases)” analysis unravels the novel therapeutic indications of c-MET and PI3K/Akt inhibitors for the treatment of glioblastoma, and potentially other cancers, in the hypoxic tumor microenvironment.

Nanoparticle-induced chemoresistance: the emerging modulatory effects of engineered nanomaterials on human intestinal cancer cell redox metabolic adaptation

“…Ingenuity pathway analysis (IPA) (Qiagen Inc., Germany), gene set enrichment analysis and network analysis were performed as reported. 48…”

“…Ingenuity pathway analysis (IPA) (Qiagen Inc., Germany) gene set enrichment analysis and network analysis was performed as reported. 48 Table 1. Sequence of gene-specific primer pairs used for RT-PCR Thereafter, the cells were harvested via trypsinization and centrifugation (1,500 rpm for 3 min), followed by a PBS wash.…”

Protein phosphorylation: A potential target in glioma development

“…MDA‐9 suppresses high levels of autophagy by phosphorylating BCL2 and EGFR signaling to achieve self‐protection 45,46 . Besides, regarding autophagy, PAK1‐mediated phosphorylation of ATG5 and phosphorylation of c‐MET and PI3K in GPCs under hypoxia conditions could affect cell autophagy 47,48 . There was an experiment that revealed that the phosphorylation level of p62/SQSTM1 was reduced in the presence of PIM knockdown.…”

Design, Synthesis and Antitumor Activity of Novel Selenium-Containing Tepotinib Derivatives as Dual Inhibitors of c-Met and TrxR

“…It has been reported that tepotinib, a potent c-Met inhibitor, exhibits promising activity in advanced HCC with c-Met overexpression in clinical studies, indicating that c-Met may serve as a therapeutic target for HCC [ 5 ]. Recent studies have demonstrated that activation of c-Met can modulate the redox protective nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) and downregulate reactive oxygen species (ROS), eventually inhibiting the death of cancer cells [ 6 ]. Treatment combined with c-Met and HO-1 inhibitors can promote ROS-induced oxidative stress and markedly reduce tumor growth [ 7 ].…”