Specificity of Phosphorylation Responses to Mitogen Activated Protein (MAP) Kinase Pathway Inhibitors in Melanoma Cells

Basken, Joel; Stuart, Scott A.; Kavran, Andrew J; Lee, Thomas; Ebmeier, Christopher C.; Old, William M.; Ahn, Natalie G.

doi:10.1074/mcp.ra117.000335

Cited by 30 publications

(29 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Knowledge on the MM phosphoproteome was generated by applying phosphopeptide enrichment protocols and LC-MS/MS on MM-derived cell lines (Basken et al 2018;Galan et al 2014;Smit et al 2014). Amongst the methodologies available to enrich phosphopeptides in MM, the most widely practiced are immobilised metal ion chromatography (IMAC) (Thingholm and Larsen 2016a) and titanium dioxide (Thingholm and Larsen 2016b), and combinations thereof.…”

Section: Pathway Signalling and Protein Phosphorylationmentioning

confidence: 99%

Clinical protein science in translational medicine targeting malignant melanoma

et al. 2019

View full text Add to dashboard Cite

Melanoma of the skin is the sixth most common type of cancer in Europe and accounts for 3.4% of all diagnosed cancers. More alarming is the degree of recurrence that occurs with approximately 20% of patients lethally relapsing following treatment. Malignant melanoma is a highly aggressive skin cancer and metastases rapidly extend to the regional lymph nodes (stage 3) and to distal organs (stage 4). Targeted oncotherapy is one of the standard treatment for progressive stage 4 melanoma, and BRAF inhibitors (e.g. vemurafenib, dabrafenib) combined with MEK inhibitor (e.g. trametinib) can effectively counter BRAFV600E-mutated melanomas. Compared to conventional chemotherapy, targeted BRAFV600E inhibition achieves a significantly higher response rate. After a period of cancer control, however, most responsive patients develop resistance to the therapy and lethal progression. The many underlying factors potentially causing resistance to BRAF inhibitors have been extensively studied. Nevertheless, the remaining unsolved clinical questions necessitate alternative research approaches to address the molecular mechanisms underlying metastatic and treatment-resistant melanoma. In broader terms, proteomics can address clinical questions far beyond the reach of genomics, by measuring, i.e. the relative abundance of protein products, post-translational modifications (PTMs), protein localisation, turnover, protein interactions and protein function. More specifically, proteomic analysis of body fluids and tissues in a given medical and clinical setting can aid in the identification of cancer biomarkers and novel therapeutic targets. Achieving this goal requires the development of a robust and reproducible clinical proteomic platform that encompasses automated biobanking of patient samples, tissue sectioning and histological examination, efficient protein extraction, enzymatic digestion, mass spectrometry-based quantitative protein analysis by label-free or labelling technologies and/or enrichment of peptides with specific PTMs. By combining data from, e.g. phosphoproteomics and acetylomics, the protein expression profiles of different melanoma stages can provide a solid framework for understanding the biology and progression of the disease. When complemented by proteogenomics, customised protein sequence databases generated from patient-specific genomic and transcriptomic data aid in interpreting clinical proteomic biomarker data to provide a deeper and more comprehensive molecular characterisation of cellular

show abstract

Section: Pathway Signalling and Protein Phosphorylationmentioning

confidence: 99%

Clinical protein science in translational medicine targeting malignant melanoma

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Neither inhibitor significantly affected k 2 . GDC-0994 does not alter phosphorylation of cellular ERK1/2 (26), and neither it nor AMP-PNP affected MKP3-catalyzed dephosphorylation of 2P-ERK2 (…”

Section: Inhibitor Binding Modulates Erk Dephosphorylationmentioning

confidence: 94%

“…We considered the possibility that conformation-selective inhibitors alter the dynamics of the activation loop in a manner that affects regulatory mechanisms for ERK2. Previous studies have reported effects of Vertex-11e and SCH772984 on the steady state phosphorylation of ERK1/2 in human cancer cells (17,24,26). Often, SCH772984 treatment suppresses dual phosphorylation of ERK1/2, while Vertex-11e increases phosphorylation, without affecting the activity state of MKK1/2.…”

Section: Inhibitor Binding Modulates Erk Dephosphorylationmentioning

confidence: 99%

Activation loop dynamics are controlled by conformation-selective inhibitors of ERK2

Pegram

Liddle

Xiao

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Modulating the dynamics of protein kinases expands the inhibitory mechanisms for small molecules. NMR measurements of the MAP kinase, ERK2, have shown that activation by dualphosphorylation induces global motions involving exchange between two states, "L'' and "R".We show that ERK inhibitors Vertex-11e and SCH772984 exploit the small energetic difference between L and R to shift the equilibrium in opposing directions, while inhibitor GDC-0994 and ATP analogue AMP-PNP retain L⇌R exchange. An X-ray structure of active 2P-ERK2 complexed with AMP-PNP reveals a shift in the Gly-rich loop along with domain closure to position the nucleotide in a more catalytically productive conformation relative to inactive 0P-ERK2:ATP. X-ray structures of 2P-ERK2 complexed with Vertex-11e or GDC-0994 recapitulate this closure, which is blocked in a complex with a SCH772984 analogue. Thus, the L R shift in 2P-ERK2 is associated with movements needed to form a competent active site. Solution measurements by hydrogen-exchange mass spectrometry (HX-MS) reveal distinct binding modes for Vertex-11e, GDC-0994 and AMP-PNP to active vs inactive ERK2, where the extent of HX protection matches their degree of R-state formation. In addition, Vertex-11e and SCH772984 show opposite effects on HX near the activation loop, suggesting that L⇌R exchange involves coupling between the activation loop and the active site. Consequently, these inhibitors differentially affect MAP kinase phosphatase activity towards 2P-ERK2. We conclude that global motions in ERK2 promote productive nucleotide binding, and couple with the activation loop to allow control of dephosphorylation by conformation-selective inhibitors.

show abstract

“…In recent years, mass spectrometry (MS)-based phosphoproteomics has provided an invaluable tool for the comprehensive discovery of signaling networks involved in drug resistance. Following the treatment of B-RAF mutant melanoma with different mitogen-activated protein kinase (MAPK) pathway inhibitors, a stable isotope labeling by amino acids in cell culture (SILAC)-based phosphoproteomic analysis revealed the inhibitor-specific regulation of MAPK kinase 1/2 or extracellular signal-regulated kinase 1/2 and an off-target reaction with p38α and led to the identification of several novel targets in the B-RAFdriven pathway 13 . Similarly, quantitative phosphoproteomic analysis of secondary resistant gastrointestinal stroma tumors helped to identify several bypassing KIT downstream elements and the simultaneous activation of EGFR 14 .…”

Section: Introductionmentioning

confidence: 99%

Quantitative phosphoproteomic analysis identifies the potential therapeutic target EphA2 for overcoming sorafenib resistance in hepatocellular carcinoma cells

Chen

Liao

et al. 2020

Exp Mol Med

View full text Add to dashboard Cite

Limited therapeutic options are available for advanced-stage hepatocellular carcinoma owing to its poor diagnosis. Drug resistance to sorafenib, the only available targeted agent, is commonly reported. The comprehensive elucidation of the mechanisms underlying sorafenib resistance may thus aid in the development of more efficacious therapeutic agents. To clarify the signaling changes contributing to resistance, we applied quantitative phosphoproteomics to analyze the differential phosphorylation changes between parental and sorafenib-resistant HuH-7 cells. Consequently, an average of~1500 differential phosphoproteins were identified and quantified, among which 533 were significantly upregulated in resistant cells. Further bioinformatic integration via functional categorization annotation, pathway enrichment and interaction linkage analysis led to the discovery of alterations in pathways associated with cell adhesion and motility, cell survival and cell growth and the identification of a novel target, EphA2, in resistant HuH-7 R cells. In vitro functional analysis indicated that the suppression of EphA2 function impairs cell proliferation and motility and, most importantly, overcomes sorafenib resistance. The attenuation of sorafenib resistance may be achieved prior to its development through the modulation of EphA2 and the subsequent inhibition of Akt activity. Binding analyses and in silico modeling revealed a ligand mimic lead compound, prazosin, that could abate the ligand-independent oncogenic activity of EphA2. Finally, data obtained from in vivo animal models verified that the simultaneous inhibition of EphA2 with sorafenib treatment can effectively overcome sorafenib resistance and extend the projected survival of resistant tumor-bearing mice. Thus our findings regarding the targeting of EphA2 may provide an effective approach for overcoming sorafenib resistance and may contribute to the management of advanced hepatocellular carcinoma.

show abstract

Specificity of Phosphorylation Responses to Mitogen Activated Protein (MAP) Kinase Pathway Inhibitors in Melanoma Cells

Cited by 30 publications

References 61 publications

Clinical protein science in translational medicine targeting malignant melanoma

Clinical protein science in translational medicine targeting malignant melanoma

Activation loop dynamics are controlled by conformation-selective inhibitors of ERK2

Quantitative phosphoproteomic analysis identifies the potential therapeutic target EphA2 for overcoming sorafenib resistance in hepatocellular carcinoma cells

Contact Info

Product

Resources

About