Fig. 4: Identification of pyrvinium pamoate as anti-leukemic compound active in RAS+ AML.
A Schematic representation of high-density pharmacological screen in NF1 KO TF-1 cells. B First screen using the PCL library of 1280 compounds at 10μM and CellTiter-Glo® cell viability reagent after 72 h of incubation. Results are represented for each compound (identified by a single dot) by the relation between their robust Z-score value (RZ-score) in Y-axis and the percentage of cell growth in X-axis. Compounds with a RZ-score ≤ −5 (retained for further analysis) are highlighted in red. C Second screen performed with serial dilutions of the top-60 compounds from the first screen in NF1 KO TF-1 cells. Results are presented for each compound illustrated by a dot as the correspondence between their median effective dose (ED50, represented using a Log10 scale) and drug sensitivity score (DSS). The best hits are highlighted in red, and the classical AML chemotherapies (daunorubicin and cytarabine) are highlighted in orange. D Dose-range experiments using log-dilutions (10−5 to 10−8 M) of pyrvinium pamoate in CTR (non-targeting sgRNA), NF1KO-1, NF1KO_2 and NRASG12D Ba/F3 cells (left panel) or TF-1 (right panel) cells during 48 h. * indicate culture in the presence of IL-3 (Ba/F3 cells) or GM-CSF (TF-1 cells). Results in the control condition of each compound were normalized to all control conditions across each plate.
