MEK Inhibition Reverses Aberrant Signaling in Melanoma Cells through Reorganization of NRas and BRAF in Self Nanoclusters.

Szczegóły
Abstrakt

Tytuł:: MEK Inhibition Reverses Aberrant Signaling in Melanoma Cells through Reorganization of NRas and BRAF in Self Nanoclusters.
Autorzy:: Yakovian O; Racah Institute of Physics, The Hebrew University, Jerusalem, Israel.
Sajman J; Racah Institute of Physics, The Hebrew University, Jerusalem, Israel.
Arafeh R; Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Neve-Oz Y; Racah Institute of Physics, The Hebrew University, Jerusalem, Israel.
Alon M; Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Samuels Y; Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Sherman E; Racah Institute of Physics, The Hebrew University, Jerusalem, Israel. .
Źródło:: Cancer research [Cancer Res] 2021 Mar 01; Vol. 81 (5), pp. 1279-1292. Date of Electronic Publication: 2020 Dec 21.
Typ publikacji:: Journal Article; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Publication: Baltimore, Md. : American Association for Cancer Research
Original Publication: Chicago [etc.]
MeSH Terms:: GTP Phosphohydrolases/*metabolism
Melanoma/*drug therapy
Melanoma/*metabolism
Membrane Proteins/*metabolism
Proto-Oncogene Proteins B-raf/*metabolism
Skin Neoplasms/*drug therapy
Skin Neoplasms/*metabolism
Cell Line, Tumor ; Cell Membrane/drug effects ; Cell Membrane/metabolism ; Epidermal Growth Factor/pharmacology ; Extracellular Signal-Regulated MAP Kinases/metabolism ; GTP Phosphohydrolases/genetics ; Humans ; MAP Kinase Kinase 1/antagonists & inhibitors ; MAP Kinase Kinase 1/metabolism ; Membrane Proteins/genetics ; Mutation ; Protein Kinase Inhibitors/pharmacology ; Proto-Oncogene Proteins B-raf/genetics ; Pyridones/pharmacology ; Pyrimidinones/pharmacology ; Signal Transduction/drug effects ; Single Molecule Imaging ; Melanoma, Cutaneous Malignant
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Substance Nomenclature:: 0 (Membrane Proteins)
0 (Protein Kinase Inhibitors)
0 (Pyridones)
0 (Pyrimidinones)
33E86K87QN (trametinib)
62229-50-9 (Epidermal Growth Factor)
EC 2.7.11.1 (BRAF protein, human)
EC 2.7.11.1 (Proto-Oncogene Proteins B-raf)
EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
EC 2.7.12.2 (MAP Kinase Kinase 1)
EC 3.6.1.- (GTP Phosphohydrolases)
EC 3.6.1.- (NRAS protein, human)
Entry Date(s):: Date Created: 20201223 Date Completed: 20210520 Latest Revision: 20231213
Update Code:: 20240105
DOI:: 10.1158/0008-5472.CAN-20-1205
PMID:: 33355187
: Czasopismo naukowe

Hotspot mutations of the oncogenes BRAF and NRas are the most common genetic alterations in cutaneous melanoma. Still, the nanoscale organization and signal coupling of these proteins remain incompletely understood, particularly upon expression of oncogenic NRas mutants. Here we employed single-molecule localization microscopy to study the nanoscale organization of NRas and BRAF at the plasma membrane (PM) of melanoma cells. NRas and BRAF resided in self-clusters that did not associate well in resting cells. In EGF-activated cells, NRas clusters became more diffused while overall protein levels at the PM increased; thus allowing enhanced association of NRas and BRAF and downstream signaling. In multiple melanoma cell lines, mutant NRas resided in more pronounced self-clusters relative to wild-type (WT) NRas yet associated more with the clustered and more abundant BRAF. In cells resistant to trametinib, a clinical MEK inhibitor (MEKi), a similar coclustering of NRas and BRAF was observed upon EGF activation. Strikingly, treatment of cells expressing mutant NRas with trametinib reversed the effect of mutant NRas expression by restoring the nonoverlapping self-clusters of NRas and BRAF and by reducing their PM levels and elevated pERK levels caused by mutant NRas. Our results indicate a new mechanism for signal regulation of NRas in melanoma through its nanoscale dynamic organization and a new mechanism for MEKi function in melanoma cells carrying NRas mutations but lacking MEK mutations. SIGNIFICANCE: Nanoscale dynamic organization of WT and mutant NRas relative to BRAF serves as a regulatory mechanism for NRas signaling and may be a viable therapeutic target for its sensitivity to MEKi.
(©2020 American Association for Cancer Research.)

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