Deneddylation of ribosomal proteins promotes synergy between MLN4924 and chemotherapy to elicit complete therapeutic responses
Year: 2024
Type: article
Abstract: (Cell Reports 42, 112925; August 29, 2023) In the originally published version of this paper, the concentration of inhibitors for (C)–(F), (G)–(H), and (I)–(K) were not included. The complete legend appears here and has been corrected online. The authors apologize for this oversight. Figure 4. Cross-regulation among synergy effectors drives macronucleoli and apoptosis(A) WERI-RB1 cells were transduced with control or one of two sgRNA viruses for the indicated hits, selected, treated with DMSO, 200 nM MLN, 200 nM TPT, or both, then westerns performed at 24 h to assess cullin neddylation and level of the indicated hits (6 h westerns in Figure S8A). (B) WERI-RB1 cells were transduced with sgRNA viruses and drug-treated as in (A) then nucleolar size quantified at 48 h (E) (n = 3, mean ± SD, ∗∗p < 0.01, one-way ANOVA, Tukey’s multiple comparison test). (C–F) WERI-RB1 cells were treated with DMSO, MLN, TPT, or MLT + TPT as indicated (color bars) plus vehicle or one of the indicated transcription/translation inhibitors (a–e in legend) and the effects on multiple readouts assessed. (C) PARP cleavage (representative blot above, graph of n = 3 ± SD below). (D) Cell morphology (yellow arrows highlight apoptotic cells in MLN + TPT-treated samples. Scale bars, 20 μm. (E) Nucleolar size. (F) Cell numbers defined with trypan blue counts. n = 3 ± SD, ∗∗p < 0.01, one-way ANOVA, Tukey’s multiple comparison test. Concentration of inhibitors: 0.1 μg/mL Cycloheximide, 10 μM CX5461, 10 μg/mL α-amanitin, and 0.1 μg/mL Actinomycin D (Act D). (G–H) MLN + TPT-treated WERI-RB1 cells were co-treated as indicated (a–e in legend; n = 2, mean ± range) and westerns used to determine fold change in effectors (G) or BAX/BCL2L1 ratio (H) relative to DMSO-treated cells (dotted lines). Concentration of inhibitors was the same as in (C)–(F). (I–J) Correlations between nucleolar size and cell number or apoptosis (I), or between BAX/BCL2L1 ratio and cell number, apoptosis, or nucleolar size (J) in WERI-RB1 cells treated with MLN + TPT and exposed to the indicated sgRNAs (1–5 in legend) or drugs (a–e in legend). Drug concentrations were the same as in (C)–(F). (K) WERI-RB1 cells were treated for 24 h with 200 nM MLN, 200 nM TPT, or 2 μM NAV, as single, two-, or three-drug combos as indicated, and cell number (trypan blue, red bars) and apoptosis (PARP cleavage, blue bars) quantified (n = 3, mean ± SD, ∗∗p < 0.01, one-way ANOVA, Tukey’s multiple comparison test). Representative PARP blot is shown. Deneddylation of ribosomal proteins promotes synergy between MLN4924 and chemotherapy to elicit complete therapeutic responsesAubry et al.Cell ReportsAugust 7, 2023In BriefPrior studies with MLN4924/Pevonedistat attribute efficacy in cancer treatment to inhibition of cullin neddylation. Aubry et al. show that blocking ribosomal protein neddylation underpins MLN4924 synergy with standard-of-care drugs in retinoblastoma and multiple myeloma. Complete responses to retinoblastoma treatment in vivo are achieved without harming vision. Full-Text PDF Open Access
Source: Cell Reports
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $5,460
OpenAlex ID: W4390937506