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Proteolysis targeting chimeric molecules in MM
to baseline by 24 h and BRD 4 was still less than control at 48 h. For KMS28BM, levels returned to baseline for BRD 2 and BRD 4 by 6 and 24 h, respectively.
ARV 825 PROTAC: CRBN expression levels correlated with levels of degradation of bromodomains 4 and MYC
We examined whether CRBN expression correlated with effect of ARV 825-mediated degradation of BRD 4 and MYC levels. The KMS11 and KMS28BM cells+over-
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expressed CRBN were treated with 10 nM and 100 nM ARV 825, respectively, for 4 h. Levels of BRD 4 and MYC RNA and protein were measured. CRBN over-expressed KMS11 and KMS28BM cells markedly decreased their RNA and protein levels of BRD 4 and MYC compared to their wild-type cells after ARV 825 treatment (Figure 3B) (KMS11 cells showed a greater decrease in BRD 4 and MYC compared to KMS28BM cells exposed to ARV 825).
Figure 7. RNA sequencing to obtain pro- file of gene expression. (A) Heatmaps show top 20 down- and up-regulated genes upon ARV 825 treatment [20 nM ARV 825, 8 hours (h)] of KMS11 MM cells. Results of two replicates from each group are shown. (B) Level of mRNA of nine genes were validated by quantitative realt-time polymerase chain reaction. (These genes were cho- sen because their expression levels were altered as viewed by RNA-sequenc- ing). Expression of each gene was nor- malized to b-actin as a reference (con- trol value converted to the value of 1). *P≤0.01; **P≤0.001; ***P≤0.0001.
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haematologica | 2019; 104(6)
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