ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma

Abstract Treatment of BRAF mutant melanomas with specific BRAF inhibitors leads to tumor remission. However, most patients eventually relapse due to drug resistance. Therefore, we designed an integrated strategy using (phospho)proteomic and functional genomic platforms to identify drug targets whose...

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Main Authors: Marjon A Smit, Gianluca Maddalo, Kylie Greig, Linsey M Raaijmakers, Patricia A Possik, Bas van Breukelen, Salvatore Cappadona, Albert JR Heck, AF Maarten Altelaar, Daniel S Peeper
Format: Article
Language:English
Published: Springer Nature 2014-12-01
Series:Molecular Systems Biology
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Online Access:https://doi.org/10.15252/msb.20145450
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author Marjon A Smit
Gianluca Maddalo
Kylie Greig
Linsey M Raaijmakers
Patricia A Possik
Bas van Breukelen
Salvatore Cappadona
Albert JR Heck
AF Maarten Altelaar
Daniel S Peeper
author_facet Marjon A Smit
Gianluca Maddalo
Kylie Greig
Linsey M Raaijmakers
Patricia A Possik
Bas van Breukelen
Salvatore Cappadona
Albert JR Heck
AF Maarten Altelaar
Daniel S Peeper
author_sort Marjon A Smit
collection DOAJ
description Abstract Treatment of BRAF mutant melanomas with specific BRAF inhibitors leads to tumor remission. However, most patients eventually relapse due to drug resistance. Therefore, we designed an integrated strategy using (phospho)proteomic and functional genomic platforms to identify drug targets whose inhibition sensitizes melanoma cells to BRAF inhibition. We found many proteins to be induced upon PLX4720 (BRAF inhibitor) treatment that are known to be involved in BRAF inhibitor resistance, including FOXD3 and ErbB3. Several proteins were down‐regulated, including Rnd3, a negative regulator of ROCK1 kinase. For our genomic approach, we performed two parallel shRNA screens using a kinome library to identify genes whose inhibition sensitizes to BRAF or ERK inhibitor treatment. By integrating our functional genomic and (phospho)proteomic data, we identified ROCK1 as a potential drug target for BRAF mutant melanoma. ROCK1 silencing increased melanoma cell elimination when combined with BRAF or ERK inhibitor treatment. Translating this to a preclinical setting, a ROCK inhibitor showed augmented melanoma cell death upon BRAF or ERK inhibition in vitro. These data merit exploration of ROCK1 as a target in combination with current BRAF mutant melanoma therapies.
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spelling doaj-art-29adb95d2eb0448bb24189c0b9582da92025-08-20T03:46:34ZengSpringer NatureMolecular Systems Biology1744-42922014-12-01101211510.15252/msb.20145450ROCK1 is a potential combinatorial drug target for BRAF mutant melanomaMarjon A Smit0Gianluca Maddalo1Kylie Greig2Linsey M Raaijmakers3Patricia A Possik4Bas van Breukelen5Salvatore Cappadona6Albert JR Heck7AF Maarten Altelaar8Daniel S Peeper9Division of Molecular Oncology, The Netherlands Cancer InstituteBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht UniversityDivision of Molecular Oncology, The Netherlands Cancer InstituteBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht UniversityDivision of Molecular Oncology, The Netherlands Cancer InstituteBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht UniversityBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht UniversityBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht UniversityBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht UniversityDivision of Molecular Oncology, The Netherlands Cancer InstituteAbstract Treatment of BRAF mutant melanomas with specific BRAF inhibitors leads to tumor remission. However, most patients eventually relapse due to drug resistance. Therefore, we designed an integrated strategy using (phospho)proteomic and functional genomic platforms to identify drug targets whose inhibition sensitizes melanoma cells to BRAF inhibition. We found many proteins to be induced upon PLX4720 (BRAF inhibitor) treatment that are known to be involved in BRAF inhibitor resistance, including FOXD3 and ErbB3. Several proteins were down‐regulated, including Rnd3, a negative regulator of ROCK1 kinase. For our genomic approach, we performed two parallel shRNA screens using a kinome library to identify genes whose inhibition sensitizes to BRAF or ERK inhibitor treatment. By integrating our functional genomic and (phospho)proteomic data, we identified ROCK1 as a potential drug target for BRAF mutant melanoma. ROCK1 silencing increased melanoma cell elimination when combined with BRAF or ERK inhibitor treatment. Translating this to a preclinical setting, a ROCK inhibitor showed augmented melanoma cell death upon BRAF or ERK inhibition in vitro. These data merit exploration of ROCK1 as a target in combination with current BRAF mutant melanoma therapies.https://doi.org/10.15252/msb.20145450kinome shRNA genomic screeningPLX4720proteomicsROCK1
spellingShingle Marjon A Smit
Gianluca Maddalo
Kylie Greig
Linsey M Raaijmakers
Patricia A Possik
Bas van Breukelen
Salvatore Cappadona
Albert JR Heck
AF Maarten Altelaar
Daniel S Peeper
ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma
Molecular Systems Biology
kinome shRNA genomic screening
PLX4720
proteomics
ROCK1
title ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma
title_full ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma
title_fullStr ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma
title_full_unstemmed ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma
title_short ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma
title_sort rock1 is a potential combinatorial drug target for braf mutant melanoma
topic kinome shRNA genomic screening
PLX4720
proteomics
ROCK1
url https://doi.org/10.15252/msb.20145450
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AT linseymraaijmakers rock1isapotentialcombinatorialdrugtargetforbrafmutantmelanoma
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