Figure 3: MiSL identifies a novel therapeutic target for the IDH1 mutation in acute myeloid leukaemia. | Nature Communications

Figure 3: MiSL identifies a novel therapeutic target for the IDH1 mutation in acute myeloid leukaemia.

From: Systematic discovery of mutation-specific synthetic lethals by mining pan-cancer human primary tumor data

Figure 3

(a) Schematic for testing druggable targets for IDH1 R132H mutation in AML. MiSL candidates were filtered for druggability using the DGIdb database. Seventeen compounds were obtained that were predicted to antagonize/inhibit the products of these genes, which were tested for synthetic lethality with IDH1 R132H. (b) Seventeen drugs were tested in the absence (−Dox) or presence (+Dox) of IDH1 R132H mutation and IC50s were calculated based on cell viability after 72 h. (c) MiSL found (i) mutual exclusion of IDH1 mutation and ACACA gene deletion, (ii) ACACA deletion causes low expression (P=0.001) and (iii) expression of ACACA is higher in IDH1-mutated AML (P=0.008). (d) Viable cell growth (PrestoBlue fluorescence) of THP-1 cells at 10 days expressing IDH1 wild type (−Dox) or R132H mutant (+Dox) plated in 2 μM TOFA or DMSO in 0.5% serum, bars show s.d., P value=0.0001. (e) Purified primary IDH1 mutant and IDH1 wild-type AML blasts plated in increasing concentrations of TOFA. Sigmoidal dose–response IC50s were calculated and compared with Mann–Whitney U, P value=0.01. (f) Viable cell growth of THP-1 cells expressing IDH1 wild type (−Dox) or R132H mutant (+Dox) transduced with scrambled or ACACA shRNA lentivirus in 0.5% serum as in d. (g) Viable cell growth of THP-1-inducible IDH1 wild-type or R132H (+/−Dox) cells transduced with lentiviral pLENTICRISPR v2 targeting ACACA exon 4 versus non-ACACA targeted controls. (h) Growth of single clones from the same pLENTICRISPR v2 transduced THP-1 cells as in g. (i) Primary IDH1 R132 mutant AML blasts were transduced with ACACA shRNA #1 or scrambled shRNA and transplanted into immunodeficient NSG mice. Bone marrow at 12 weeks was analysed for human CD45+CD33+ RFP+ cells. Scatter plot shows absolute and average percentage of hCD33+ RFP+ cells gated on human CD45+ engrafted cells (n=3, non-targeting versus n=5, shRNA#1, *P<0.05, Mann–Whitney). Panels d and f–h show representative of three independent experiments using four biological replicates; Student’s t-test is used to calculate significance with P values as shown.

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