NPM1c+ AML cells are preferentially sensitive to the inhibition of TSR3 and related ribosome biogenesis factors. (A) Analysis of DepMap data was used to identify the top 500 preferentially essential genes for the OCI-AML3 cell line. Of these, candidates with an established role in ribosome biogenesis are highlighted (right). The “gene_effect” represents the dependency score of each candidate for OCI-AML3 cells, where a score of 0 indicates that the gene is not essential, whereas a score of −1 corresponds to the median of all common essential genes in the database. The mean represents the mean gene effect across all cancer cell lines in the database. (B-D) Differential dependency of 37 AML cell lines to CRISPR knockout of TSR1 (B), BYSL (C), or TSR3 (D) according to DepMap. (E) Venn diagram of gene dropouts in a CRISPR screen with 5 AML cell lines (OCI-AML3, OCI-AML2, MOLM-13, HL-60, and MV-4-11).23 (F-H) Relative proliferation of OCI-AML3 (F), murine Npm1cA/+; Flt3ITD/+AML (G), or nonleukemic 32D (H) cells on knockout of TSR3 as measured by a competitive proliferation assay of gRNA-BFP positive (BFP+) transduced cells vs untransduced BFP negative (BFP−) cells in 15 to 18 days. BFP percentage was normalized to the day 4 reading and for panel G to the “gControl” (mean ± SD, n = 3 independent experiments). (F-G) t test was used to calculate P values between groups. ∗∗P ≤ .01; ∗∗∗P ≤ .001; ∗∗∗∗P ≤ .0001. (A-D) CRISPR scores originate from the DepMap Public 23Q2+Score, Chronos data set.24,25 Panel A was created with BioRender.com. Vassiliou, G. (2025) https://BioRender.com/9tt8kww.
