Activating mutations in NRAS account for 15–20% of melanoma, yet effective anti-NRAS therapies are still lacking. In this study, we unveil the casein kinase 1δ (CK1δ) as an uncharacterized regulator of oncogenic NRAS mutations, specifically Q61R and Q61K, which are the most prevalent NRAS mutations in melanoma. The genetic ablation or pharmacological inhibition of CK1δ markedly destabilizes NRAS mutants and suppresses their oncogenic functions. Moreover, we identify USP46 as a bona fide deubiquitinase of NRAS mutants. Mechanistically, CK1δ directly phosphorylates USP46 and activates its deubiquitinase activity towards NRAS mutants, thus promoting oncogenic NRAS-driven melanocyte malignant transformation and melanoma progression in vitro and in vivo. Our findings underscore the significance of the CK1δ-USP46 axis in stabilizing oncogenic NRAS mutants and provide preclinical evidence that targeting this axis holds promise as a therapeutic strategy for human melanoma harboring NRAS mutations.
Highly accurate analysis of single-nucleotide polymorphisms (SNPs) plays an important role in both disease diagnostics and personalized medicine development. In this work, a DNA microarray-based resonance light scattering (RLS) assay has been developed for multiplexed detection of papillary thyroid carcinoma (PTC) related mutation points including BRAFV600E (t1m), NRAS codon 61 (t2m), TERT promoter g.1295228 (t31m) and TERT promoter g.1295250 (t32m) with high sensitivity and selectivity by the attachment of polyvalent ssDNA modified 13 nm gold nanoparticles (ssDNAs@GNPs) followed by silver deposition for signal enhancement. The microarray-based RLS assay provides a detection limit (S/N = 3) at the sub-nanomolar level for the target ssDNAs and determines allele frequencies as low as 0.2% for t1m, 0.2% for t2m, 0.5% for t31m, and 0.5% for t32m in the cocktail of target ssDNAs, respectively. The practicability of the DNA microarray-based RLS assay is demonstrated by profiling of t2m in 50 clinical thyroid tissue samples of PTC patients, and satisfactory results are obtained.
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