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Transcriptome analysis of human lung andenocarcinoma cell line A549, in which ARAF was depleted and reconstituted with an empty vector control, ARAF wildtype, or a dimer deficient and kinase inactive mutant.

ABSTRACT: Transcriptome analysis of human lung andenocarcinoma cell line A549, in which ARAF was depleted and reconstituted with an empty vector control, ARAF wildtype, or a dimer deficient and kinase inactive mutant.

PROVIDER: PRJNA629473 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11650300.fastq.gz	Fastqsanger.gz
	SRR11650301.fastq.gz	Fastqsanger.gz
	SRR11650296.fastq.gz	Fastqsanger.gz
	SRR11650297.fastq.gz	Fastqsanger.gz
	SRR11650298.fastq.gz	Fastqsanger.gz

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Transcriptome analysis of human lung andenocarcinoma cell line A549, in which ARAF was depleted and reconstituted with an empty vector control, ARAF wildtype, or a dimer deficient and kinase inactive mutant.

Project description:RNA sequencing analysis of A549 cells that were depleted for ARAF (shARAF) and reconstituted with either empty vector control (+EV) or ARAF wildtype (+ARAF) or dimer deficient ARAF kinase (+R362H) respectively to identify factors that are differentially regulated by ARAF and its kinase activity. In particular, we discovered that ARAF regulates genes that are involved in proliferation and malignant growth by surpressing gene expression of members of the ERBB3/ AKT signaling pathway. At the same time, a functional interpretation of the differentially expressed (DE) genes with topGO indicated an important role of ARAF in regulation of genes involved in, among others, cell adhesion, cell motility, epithelial cell migration.

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ARAF Mutations Confer Resistance to RAF Dimer Inhibitor Belvarafenib in NRAS- and BRAF- Mutant Melanoma

Project description:Belvarafenib, a potent and selective RAF dimer (type II) inhibitor, exhibits clinical activity in BRAFV600E- and NRAS-mutant melanoma patients. Here, we report the first-in-human phase I study investigating maximum tolerated dose, assessing safety and preliminary efficacy of belvarafenib in BRAF- and RAS-mutated advanced solid tumors (NCT02405065, NCT03118817). Through generation of belvarafenib resistant NRAS- mutant melanoma cells and analysis of circulating tumor DNA from patients treated with belvarafenib, we identified novel recurrent mutations in ARAF within the kinase domain. ARAF mutants conferred resistance to belvarafenib in both a dimer- and kinase activity- dependent manner. Belvarafenib induced ARAF mutant dimers, and dimers containing mutant ARAF were active in the presence of inhibitor. ARAF mutations may serve as a general resistance mechanism for RAF dimer inhibitors as the mutants exhibit reduced sensitivity to a panel of type II RAF inhibitors. The combination of RAF plus MEK inhibition may be used to delay ARAF-driven resistance and suggests a rational combination for clinical use. Taken together, our findings reveal specific and compensatory functions for the ARAF isoform and implicate ARAF mutations as a driver of resistance to RAF dimer inhibitors.

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