Genome-scale CRISPR screen reveals neddylation to contribute to cisplatin resistance of testicular germ cell tumours

Menée à l'aide de lignées cellulaires de tumeur germinale testiculaire de type II et à l'aide d'une technique de criblage génomique utilisant la technonologie d'édition CRISPR/Cas9, cette étude met en évidence l'intérêt d'inhiber NAE1, une protéine impliquée dans l'activation de la neddylation, pour lever la résistance des cellules cancéreuses au cisplatine

Résumé en anglais

Background : Type II testicular germ cell tumours (TGCT) are the most prevalent tumours in young men. Patients suffering from cisplatin-resistant TGCTs are facing very poor prognosis demanding novel therapeutic options. Neddylation is a known posttranslational modification mediating many important biological processes, including tumorigenesis. Overactivation of the neddylation pathway promotes carcinogenesis and tumour progression in various entities by inducing proteasomal degradation of tumour suppressors (e.g., p21, p27).

Methods : We used a genome-scale CRISPR/Cas9 activation screen to identify cisplatin resistance factors. TGCT cell lines were treated with the neddylation inhibitor (MLN4924)/cisplatin/combination and investigated for changes in viability (XTT assay), apoptosis/cell cycle (flow cytometry) as well as in the transcriptome (3’mRNA sequencing).

Results : NAE1 overexpression was detected in cisplatin-resistant colonies from the CRISPR screen. Inhibition of neddylation using MLN4924 increased cisplatin cytotoxicity in TGCT cell lines and sensitised cisplatin-resistant cells towards cisplatin. Apoptosis, G2/M-phase cell cycle arrest,