Global profiling of AMG510 modified proteins identified tumor suppressor KEAP1 as an off-target

Wang, Yini; Zhong, Bowen; Xu, Caixia; Zhan, Dongdong; Zhao, Songhao; Wu, Hongxing; Liu, Mingwei; Lan, Xiaoling; Cai, Danni; Ding, Qi; Zheng, Bo; Lan, Jiong; Lu, Qiang; Wang, Yi; Qin, Jun

doi:10.1016/j.isci.2023.106080

Cited by 9 publications

(10 citation statements)

References 60 publications

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“…Candidates for mediating this effect detected in our screen include PIK3R1/2, G-coupled receptor proteins, and ILK, which might contribute to the unspecific activation of AKT in these cells. It is worth noting our analysis shows that some of the changes shown in the interactomes of KRASG12C and KRAS WT upon Sotorasib treatment can be explained by the direct effect of Sotorasib on some of the proteins identified which are shown to be targeted by this drug in previous studies (i.e., ACTB, VIM or RAN) [ 14 ].…”

Section: Discussionmentioning

confidence: 87%

“…For this, we monitored the effects of the KRASG12C mutant inhibitor, Sotorasib, on the regulation of the best characterised RAS effector pathways, RAF and AKT, in cell lines that express different KRAS mutants. Sotorasib inhibits KRASG12C at nM concentrations but in different studies is used at a wide range of concentrations [ 14 , 52 ]. Importantly, different groups are using high concentrations of Sotorasib (5–10 µM) to generate resistant cell lines that can be used to characterise the mechanisms of resistance to this drug [ 53 , 54 , 55 ].…”

Section: Resultsmentioning

confidence: 99%

“…Although Sotorasib is approved as a KRASG12C-specific inhibitor, different groups have identified other proteins that bind this drug and we reasoned that some of these targets might also be part of the KRAS interactome [ 14 , 52 , 60 , 61 ]. To explore this, we used information from one of the studies that have identified proteins that are targeted by Sotorasib using an MS-based approach [ 14 ]. This study identified 183 proteins that bind Sotorasib and using a Venn diagram analysis we saw that 15 Sotorasib target proteins were identified in our screen as part of the interactome of KRAS proteins ( Figure 7 C).…”

Section: Resultsmentioning

confidence: 99%

“…Unfortunately, while the initial response rates to Sotorasib and Adagrasib are promising, ranging from 32% to 46%, respectively, in non-small cell lung cancer (NSCLC), resistance inevitably develops in these patients [ 13 ]. These drugs can also lead to severe side effects in the patients and more information is necessary to fully characterise their mechanism of action [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

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Proteomic Mapping of the Interactome of KRAS Mutants Identifies New Features of RAS Signalling Networks and the Mechanism of Action of Sotorasib

Nolan,

Raso,

Kolch

et al. 2023

Cancers

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RAS proteins are key regulators of cell signalling and control different cell functions including cell proliferation, differentiation, and cell death. Point mutations in the genes of this family are common, particularly in KRAS. These mutations were thought to cause the constitutive activation of KRAS, but recent findings showed that some mutants can cycle between active and inactive states. This observation, together with the development of covalent KRASG12C inhibitors, has led to the arrival of KRAS inhibitors in the clinic. However, most patients develop resistance to these targeted therapies, and we lack effective treatments for other KRAS mutants. To accelerate the development of RAS targeting therapies, we need to fully characterise the molecular mechanisms governing KRAS signalling networks and determine what differentiates the signalling downstream of the KRAS mutants. Here we have used affinity purification mass-spectrometry proteomics to characterise the interactome of KRAS wild-type and three KRAS mutants. Bioinformatic analysis associated with experimental validation allows us to map the signalling network mediated by the different KRAS proteins. Using this approach, we characterised how the interactome of KRAS wild-type and mutants is regulated by the clinically approved KRASG12C inhibitor Sotorasib. In addition, we identified novel crosstalks between KRAS and its effector pathways including the AKT and JAK-STAT signalling modules.

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Section: Discussionmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Proteomic Mapping of the Interactome of KRAS Mutants Identifies New Features of RAS Signalling Networks and the Mechanism of Action of Sotorasib

Nolan,

Raso,

Kolch

et al. 2023

Cancers

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“…Since sotorasib is reported to bind to >300 proteins, we ruled out non-specific effects of sotorasib on autophagy, by treating a KRAS G12V -driven NSCLC cell line (COR-L23) with sotorasib, which failed to show any change in mCherry:EGFP fluorescence (Fig. S1E) 35 . Moreover, as predicted by previous experiments, all KRAS G12X -driven NSCLC cell lines displayed a dose-dependent increase in autophagy following treatment with trametinib, a MEK1/2 inhibitor (Figs.…”

Section: Human Kras G12c -Driven Lung Cancer Cells Are Sensitive To C...mentioning

confidence: 99%

Inhibition of ULK1/2 and KRAS^G12Ccontrols tumor growth in preclinical models of lung cancer

Ghazi,

O’Toole,

Srinivas Boggaram

et al. 2024

Preprint

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Mutational activation ofKRASoccurs commonly in lung carcinogenesis and, with the recent FDA approval of covalent inhibitors of KRASG12Csuch as sotorasib or adagrasib, KRAS oncoproteins are important pharmacological targets in non-small cell lung cancer (NSCLC). However, not all KRASG12C-driven NSCLCs respond to these inhibitors, and the emergence of drug resistance in those patients that do respond can be rapid and pleiotropic. Hence, based on a backbone of covalent inhibition of KRASG12C, efforts are underway to develop effective combination therapies. Here we report that inhibition of KRASG12Csignaling increases autophagy in KRASG12Cexpressing lung cancer cells. Moreover, the combination of DCC-3116, a selective ULK1/2 inhibitor, plus sotorasib displays cooperative/synergistic suppression of human KRASG12C-driven lung cancer cell proliferationin vitroand superior tumor controlin vivo. Additionally, in genetically engineered mouse models of KRASG12C-driven NSCLC, inhibition of either KRASG12Cor ULK1/2 decreases tumor burden and increases mouse survival. Consequently, these data suggest that ULK1/2-mediated autophagy is a pharmacologically actionable cytoprotective stress response to inhibition of KRASG12Cin lung cancer.

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KRAS-mutant non-small cell lung cancer (NSCLC) therapy based on tepotinib and omeprazole combination

Rosell,

Jantus-Lewintre,

Cao

et al. 2024

Cell Commun Signal

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Background KRAS-mutant non-small cell lung cancer (NSCLC) shows a relatively low response rate to chemotherapy, immunotherapy and KRAS-G12C selective inhibitors, leading to short median progression-free survival, and overall survival. The MET receptor tyrosine kinase (c-MET), the cognate receptor of hepatocyte growth factor (HGF), was reported to be overexpressed in KRAS-mutant lung cancer cells leading to tumor-growth in anchorage-independent conditions. Methods Cell viability assay and synergy analysis were carried out in native, sotorasib and trametinib-resistant KRAS-mutant NSCLC cell lines. Colony formation assays and Western blot analysis were also performed. RNA isolation from tumors of KRAS-mutant NSCLC patients was performed and KRAS and MET mRNA expression was determined by real-time RT-qPCR. In vivo studies were conducted in NSCLC (NCI-H358) cell-derived tumor xenograft model. Results Our research has shown promising activity of omeprazole, a V-ATPase-driven proton pump inhibitor with potential anti-cancer properties, in combination with the MET inhibitor tepotinib in KRAS-mutant G12C and non-G12C NSCLC cell lines, as well as in G12C inhibitor (AMG510, sotorasib) and MEK inhibitor (trametinib)-resistant cell lines. Moreover, in a xenograft mouse model, combination of omeprazole plus tepotinib caused tumor growth regression. We observed that the combination of these two drugs downregulates phosphorylation of the glycolytic enzyme enolase 1 (ENO1) and the low-density lipoprotein receptor-related protein (LRP) 5/6 in the H358 KRAS G12C cell line, but not in the H358 sotorasib resistant, indicating that the effect of the combination could be independent of ENO1. In addition, we examined the probability of recurrence-free survival and overall survival in 40 early lung adenocarcinoma patients with KRAS G12C mutation stratified by KRAS and MET mRNA levels. Significant differences were observed in recurrence-free survival according to high levels of KRAS mRNA expression. Hazard ratio (HR) of recurrence-free survival was 7.291 (p = 0.014) for high levels of KRAS mRNA expression and 3.742 (p = 0.052) for high MET mRNA expression. Conclusions We posit that the combination of the V-ATPase inhibitor omeprazole plus tepotinib warrants further assessment in KRAS-mutant G12C and non G12C cell lines, including those resistant to the covalent KRAS G12C inhibitors.

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Global profiling of AMG510 modified proteins identified tumor suppressor KEAP1 as an off-target

Cited by 9 publications

References 60 publications

Proteomic Mapping of the Interactome of KRAS Mutants Identifies New Features of RAS Signalling Networks and the Mechanism of Action of Sotorasib

Proteomic Mapping of the Interactome of KRAS Mutants Identifies New Features of RAS Signalling Networks and the Mechanism of Action of Sotorasib

Inhibition of ULK1/2 and KRAS^G12Ccontrols tumor growth in preclinical models of lung cancer

KRAS-mutant non-small cell lung cancer (NSCLC) therapy based on tepotinib and omeprazole combination

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Global profiling of AMG510 modified proteins identified tumor suppressor KEAP1 as an off-target

Cited by 9 publications

References 60 publications

Proteomic Mapping of the Interactome of KRAS Mutants Identifies New Features of RAS Signalling Networks and the Mechanism of Action of Sotorasib

Proteomic Mapping of the Interactome of KRAS Mutants Identifies New Features of RAS Signalling Networks and the Mechanism of Action of Sotorasib

Inhibition of ULK1/2 and KRASG12Ccontrols tumor growth in preclinical models of lung cancer

KRAS-mutant non-small cell lung cancer (NSCLC) therapy based on tepotinib and omeprazole combination

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Inhibition of ULK1/2 and KRAS^G12Ccontrols tumor growth in preclinical models of lung cancer