Proteomic and Phosphoproteomic Analyses Reveal the Oncogenic Role of PTK7-NDRG1 Axis in Non-small-cell Lung Cancer Cell Resistance to AZD9291

Abstract

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are the most important chemotherapeutics for non-small-cell lung cancer (NSCLC) therapy. The resistance to EGFR-TKIs is one of the biggest obstacles to NSCLC outcome. In this study, taking advantage of phospho- and proximal proteomic techniques, we analyzed the network rearrangement in cell lines responding to AZD9291 treatment and found that cell-cell adhesion was dramatically enhanced in AZD9291-resistant cells. Further analysis revealed that protein tyrosine kinase 7 (PTK7) expression was significantly elevated. Knockdown or overexpression assays showed that PTK7 played a critical role in improving cell adhesion, which enhanced drug resistance. Because PTK7 is a membrane-localized pseudokinase, the proximal labeling probe BirA* was fused to reveal PTK7-interacting proteins. We found that PTK7 interacted with and stabilized NDRG1, which is located predominantly adjacent to adherens junctions. Downregulation of PTK7 or NDRG1 eliminated the resistance of H1975-resistant (H1975-R) and PC9-resistant (PC9-R) cells to AZD9291, suggesting that the PTK7-NDRG1 axis might be a potential target to eliminate the EGFR-TKI resistance during NSCLC therapy.

An oncogene addiction phosphorylation signature and its derived scores inform tumor responsiveness to targeted therapies

“…Many of these efforts focused solely on tyrosine phosphorylations [ 86 – 88 ] but in some of the data-dependent acquisition (DDA)-based studies modulations of particular OAPS phosphosites were detected. These included for example TERF2 S365 in PC-9 and H1975 cells following AZD9291 treatment [ 89 ] or ACLY Ser455 and NCBP1 Ser22 phosphorylation in erlotinib-treated H1975 cell line [ 90 ]. In addition, changes in other phosphorylation sites of OAPS proteins have also been recorded, such as various phosphorylations of EIF4G1, LIMA1, ACLY, EIF4B and TERF2 upon EGFR or MET inhibition [ 41 , 89 , 90 ].…”

“…These included for example TERF2 S365 in PC-9 and H1975 cells following AZD9291 treatment [ 89 ] or ACLY Ser455 and NCBP1 Ser22 phosphorylation in erlotinib-treated H1975 cell line [ 90 ]. In addition, changes in other phosphorylation sites of OAPS proteins have also been recorded, such as various phosphorylations of EIF4G1, LIMA1, ACLY, EIF4B and TERF2 upon EGFR or MET inhibition [ 41 , 89 , 90 ]. The occurrence of some of the OAPS phosphorylations in these DDA datasets adds to the weight of our SRM-based OAPS signature.…”

Prognostic correlation between specialized capillary endothelial cells and lung adenocarcinoma

“…GSTA3 acts as a cellular defence against toxic and carcinogenic substances, and its expression in alveolar cells induces an oxidative stress response [ 61 ]. The protein encoded by the NDRG1 gene is involved in the stress response, cell growth and differentiation and has been implicated in angiogenesis and drug resistance in lung cancer [ 62 , 63 ]. The function of the NPAS1 gene is still unclear, and deletion polymorphisms have been found only in breast cancer [ 64 ].…”

4EBP2-regulated protein translation has a critical role in high-fat diet–induced insulin resistance in hepatocytes

“…The enrichment method of phosphorylated peptides by TiO 2 is the same as the published article of our laboratory ( 48 ). In briefly, 500 μg protein was reduced with 10 mM DTT (D477470, Aladdin) at room temperature for 45 min, then alkylated with 40 mM iodoacetamide (I131590, Aladdin) at room temperature for 45 min.…”