Drug screening identifies tazarotene and bexarotene as therapeutic agents in Multiple Sulfatase Deficiency (2023)
Figure 2

Fig. 2. Tazarotene and bexarotene increase sulfatase activities in MSD primary fibroblasts and MSD iPSC derived NPCs.

(A) ARSA activity quantification after treatment of MSD primary fibroblasts (variant FGE Gly247Arg homozygous) with increasing concentrations of tazarotene for 6 days. Data represent mean ±SD of 3-7 independent experiments (biological replicates). One-way ANOVA followed by Tukey's test for multiple comparisons. Displayed are significance levels for the next significant difference between adjacent concentrations. #### p<0.0001. Difference against 0 µM control: *** p<0.001, **** p<0.0001. See details on p-values in appendix table S10.

(B) ARSA activity quantification after treatment of MSD primary fibroblasts (variant FGE Gly247Arg homozygous) with increasing concentrations of bexarotene for 6 days. Data represent mean ±SD of 3-9 independent experiments (biological replicates). One-way ANOVA followed by Tukey's test for multiple comparisons. Displayed are significance levels for the next significant difference between adjacent concentrations. #### p<0.0001. Difference against 0 µM control: * p<0.05, **** p<0.0001. See details on p-values in appendix table S11.

(C) ARSA activity quantification after simultaneous treatment of MSD primary fibroblasts (variant FGE Gly247Arg homozygous) with increasing concentrations of tazarotene and bexarotene in a fixed combination of 1:2 for 6 days. Data represent mean ±SD of 3-6 independent experiments (biological replicates). One-way ANOVA followed by Tukey's test for multiple comparisons. Displayed are significance levels for the next significant difference between adjacent concentrations. #### p<0.0001. Difference against 0/0 µM control: ** p<0.01, **** p<0.0001. See details on p-values in appendix table S12.

(D) Analysis and quantification of a time dependent increase of ARSA activity in MSD primary fibroblasts (variant FGE Gly247Arg homozygous) simultaneously treated with 10 and 20 µM tazarotene and bexarotene, respectively, up to 21 days. Data represent mean ±SD of 3-6 independent experiments (biological replicates). One-way ANOVA followed by Tukey's test for multiple comparisons. Displayed are significance levels for the next significant difference between adjacent treatment times. #### p<0.0001. Difference against 0 days control: * p<0.05, **** p<0.0001. See details on p-values in appendix table S13.

(E) Analysis and quantification of increased sulfatase activities different to ARSA, namely ARSB, GALNS, and STS in MSD primary fibroblasts (variant FGE Gly247Arg homozygous) after 6 days of simultaneous treatment with tazarotene/bexarotene 10/20 µM. Data represent mean ±SD of 3-6 independent experiments (biological replicates). One-way ANOVA followed by Tukey's test for multiple comparisons. *** p<0.001, **** p<0.0001. See details on p-values in appendix table S14.

(F) Quantification of ARSA activities in MSD primary fibroblasts with different homozygous SUMF1 mutations (FGE Gly247Arg, FGE Gly263Val, FGE Ala279Val, FGE Arg349Trp) after 6 days of simultaneous treatment with tazarotene/bexarotene 10/20 µM. Data represent mean ±SD of 3 independent experiments (biological replicates). One-way ANOVA followed by Tukey's test for multiple comparisons. ****p<0.0001. See details on p-values in appendix table S15.

(G) Quantification of ARSA activity in MSD patient-derived iPSCs differentiated into NPCs and unaffected control NPCs controls. Simultaneous treatment with 5 µM tazarotene and 5 µM bexarotene for 4 days. Data represent mean ±SD of 6 independent experiments (biological replicates). Unpaired t-test. **** p<0.0001. See details on p-values in appendix table S16.

(H) Quantification of SGSH activity in MSD patient-derived iPSCs differentiated into NPCs and unaffected control NPCs controls. Simultaneous treatment with 5 µM tazarotene and 5 µM bexarotene for 4 days. Data represent mean ±SD of 3 independent experiments (biological replicates). Unpaired t-test. * p<0.05, *** p<0.001. See details on p-values in appendix table S17.