Effects of Activating Mutations on EGFR Cellular Protein Turnover and Amino Acid Recycling Determined Using SILAC Mass Spectrometry

Greig, Michael J.; Niessen, Sherry; Weinrich, Scott L.; Feng, Jun; Shi, Minmin; Johnson, Ted O.

doi:10.1155/2015/798936

Cited by 24 publications

(21 citation statements)

References 30 publications

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“…Generally, parameter values across RTKs for endocytosis, recycling, degradation and synthesis were consistent with published literature values for EGFR 15 , 17 , 18 , 22 , 39 , 51 and Axl endocytosis and recycling rates were similar to those estimated from a model of ligand-receptor interaction and trafficking. 33 In addition to estimated values consistent with literature reports, we find relatively narrow distributions for parameter estimates with the methodology developed here.…”

Section: Resultssupporting

confidence: 86%

“…), is the current test parameter value, is the prior distribution mean parameter value, and is the prior distribution standard deviation. k shed values for and were as calculated above, whereas the other values were estimated from the literature to be as follows: 10 −2 for k deg , 15 , 18 k end, 17 , 22 , 51 k rec 17 and 10 2 for P syn . 39 Values were based on low end estimates as observed for EGFR as it is most commonly reported for the ligand stimulated case.…”

Section: Methodsmentioning

confidence: 99%

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Systems Modeling Identifies Divergent Receptor Tyrosine Kinase Reprogramming to MAPK Pathway Inhibition

et al. 2018

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IntroductionTargeted cancer therapeutics have demonstrated more limited clinical efficacy than anticipated, due to both intrinsic and acquired drug resistance. Underlying mechanisms have been largely attributed to genetic changes, but a substantial proportion of resistance observations remain unexplained by genomic properties. Emerging evidence shows that receptor tyrosine kinase (RTK) reprogramming is a major alternative process causing targeted drug resistance, separate from genetic alterations. Hence, the contributions of mechanisms leading to this process need to be more rigorously assessed.MethodsTo parse contributions of multiple mechanisms to RTK reprogramming, we have developed a quantitative multi-receptor and multi-mechanistic experimental framework and kinetic model.ResultsWe find that RTK reprogramming mechanisms are disparate among RTKs and nodes of intervention in the MAPK pathway. Mek inhibition induces increased Axl and Her2 levels in triple negative breast cancer (TNBC) cells while Met and EGFR levels remain unchanged, with Axl and Her2 sharing re-wiring through increased synthesis and differing secondary contributing mechanisms. While three Mek inhibitors exhibited mechanistic similarity, three Erk inhibitors elicited effects different from the Mek inhibitors and from each other, with MAPK pathway target-specific effects correlating with Erk subcellular localization. Furthermore, we find that Mek inhibitor-induced RTK reprogramming occurs through both BET bromodomain dependent and independent mechanisms, motivating combination treatment with BET and Axl inhibition to overcome RTK reprogramming.ConclusionsOur findings suggest that RTK reprogramming occurs through multiple mechanisms in a MAPK pathway target-specific manner, highlighting the need for comprehensive resistance mechanism profiling strategies during pharmacological development.Electronic supplementary materialThe online version of this article (10.1007/s12195-018-0542-y) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 86%

Section: Methodsmentioning

confidence: 99%

Systems Modeling Identifies Divergent Receptor Tyrosine Kinase Reprogramming to MAPK Pathway Inhibition

et al. 2018

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“…EGFR mutation mainly focused on the deletion of exon 19 and L858R missense mutation of exon 21 (about 86%), that was relatively close to the literature's reported 90% [22]. Further analysis of the relationship between EGFR-activating mutations and clinicopathological characteristics showed that the incidence of EGFR-activating mutations in female, adenocarcinoma and non-smoking patients was significantly increased, which was consistent with previous studies [23,24]. However, although many researches have shown that EGFR mutation was common in Asian female NSCLC patients, and the treatment effectiveness of EGFR TKIs in Asian female NSCLC patients was better than that of male patients with NSCLC [25], so from the aspect of clinical sign, it seemed that EGFR TKIs had better treatment effect in Asian female patients.…”

Section: Discussionsupporting

confidence: 87%

Association of EGFR and KRAS mutations with expression of p-AKT, DR5 and DcR1 in non-small cell lung cancer

Zhao¹,

Deng²,

Lu³

et al. 2017

neo

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The activation of AKT is one of the causes of resistance to epidermal growth factor receptor (EGFR)- tyrosine kinase inhibitors (TKIs). Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) combines with related receptors to trigger apoptosis or protect the cells against TRAIL apoptosis. This research focused on the association of EGFR and KRAS mutations with expression of AKT, p-AKT, DR5 and DcR1 in non-small cell lung cancer. 82 NSCLC patients were included in the study. xTAG liquichip techonolgy (xTAG-LCT) was applied to investigate the genetic mutation of EGFR and KRAS, Quantitative Real-time PCR was used to test the mRNA expression of AKT, DR5 and DcR1 and Western Blot was applied to test the protein expression of AKT, p-AKT, DR5, and DcR1. We found that of 82 patients, 31 cases had EGFR-activating mutations, more common in female, adenocarcinoma, and non-smoker patients; 9 cases had KRAS mutations, frequently found in patients with smoking history. The expression of AKT and p-AKT correlated with staging, tumor differentiation, and lymph node metastasis. The expression of DR5 in phase III and low differentiation tumor was significantly higher than that in phase I+II and high and median differentiation tumor; the expression of DcR1 in phase III and low differentiation tumor was significantly lower than that in phase I+II and high and median differentiation tumor. Compared with EGFR and KRAS wild type, in NSCLC tissue with EGFR and KRAS mutations, the expression of AKT and p-AKT was significantly higher. These results suggest that EGFR and KRAS mutation status was associated with the expression of AKT and p-AKT. AKT, p-AKT, DR5, and DcR1 all took part in the occurrence and development of NSCLC, and may become a reference index to evaluate the prognosis of NSCLC.

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“…Note that for detecting p-EGFR, it was reported that the expression of p-EGFR could not be clearly detected at 24 h due to the short half-life of activated EGFR (~1.5–4 h) [75]; thus, we pre-incubated NSCLC cell lines with the indicated concentrations of MG3 and CDDP in serum free media for 1 h prior to stimulation of EGFR with EGF (50 ng/mL) for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Butoxy Mansonone G Inhibits STAT3 and Akt Signaling Pathways in Non-Small Cell Lung Cancers: Combined Experimental and Theoretical Investigations

Mahalapbutr

Wonganan

Chavasiri

et al. 2019

Cancers

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Epidermal growth factor receptor (EGFR) is the key molecular target for non-small cell lung cancer (NSCLC) due to its major contribution to complex signaling cascades modulating the survival of cancer cells. Targeting EGFR-mediated signaling pathways has been proved as a potential strategy for NSCLC treatment. In the present study, mansonone G (MG), a naturally occurring quinone-containing compound, and its semi-synthetic ether derivatives were subjected to investigate the anticancer effects on human NSCLC cell lines expressing wild-type EGFR (A549) and mutant EGFR (H1975). In vitro cytotoxicity screening results demonstrated that butoxy MG (MG3) exhibits the potent cytotoxic effect on both A549 (IC50 of 8.54 μM) and H1975 (IC50 of 4.21 μM) NSCLC cell lines with low toxicity against PCS201-010 normal fibroblast cells (IC50 of 21.16 μM). Western blotting and flow cytometric analyses revealed that MG3 induces a caspase-dependent apoptosis mechanism through: (i) inhibition of p-STAT3 and p-Akt without affecting upstream p-EGFR and (ii) activation of p-Erk. The 500-ns molecular dynamics simulations and the molecular mechanics combined with generalized Born surface area (MM/GBSA)-based binding free energy calculations suggested that MG3 could possibly interact with STAT3 SH2 domain and ATP-binding pocket of Akt. According to principal component analysis, the binding of MG3 toward STAT3 and Akt dramatically altered the conformation of proteins, especially the residues in the active site, stabilizing MG3 mainly through van der Waals interactions.

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Effects of Activating Mutations on EGFR Cellular Protein Turnover and Amino Acid Recycling Determined Using SILAC Mass Spectrometry

Cited by 24 publications

References 30 publications

Systems Modeling Identifies Divergent Receptor Tyrosine Kinase Reprogramming to MAPK Pathway Inhibition

Systems Modeling Identifies Divergent Receptor Tyrosine Kinase Reprogramming to MAPK Pathway Inhibition

Association of EGFR and KRAS mutations with expression of p-AKT, DR5 and DcR1 in non-small cell lung cancer

Butoxy Mansonone G Inhibits STAT3 and Akt Signaling Pathways in Non-Small Cell Lung Cancers: Combined Experimental and Theoretical Investigations

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