Fig. 4.

Lipidome analysis identified lipid metabolism defects in mutant FUS iPSC-derived OPCs. Quantification of total lipid concentrations (nmol/mg DNA) by mass spectrometry in FUS^R521H mutant OPCs (a) and FUS^P525L mutant OPCs (f) and their respective isogenic controls (N = 3 independent differentiations). Quantification of cholesterol ester (CE) and sphingolipid concentrations (SM, Cer, and HexCer) in FUS^R521H mutant OPCs (b, c) and FUS^P525L mutant OPCs (g, h) and their respective isogenic controls. Quantification of glycerophospholipid subclasses (PC, PC(O-), PC(P-), PE, PE(O-), PE(P-), PG, and PI) in FUS^R521H mutant OPCs (e) and FUS^P525L mutant OPCs (j) and their respective isogenic controls. PC to PE concentration ratio in FUS^R521H mutant OPCs (d) and FUS^P525L mutant OPCs (i) and their respective isogenic controls. PCA plot of lipidomic data in FUS^R521H mutant OPCs (k) and FUS^P525L mutant OPCs (l) and their respective isogenic controls. m, n Heatmap representing all clustered samples and color-coded normalized concentrations of the top 50 most significantly altered individual lipid species (one-way ANOVA, p < 0.05, FDR corrected). (PC, phosphatidylcholine; PC(O-), alkylphosphatidylcholine; PC(P-), alkenylphosphatidylcholine; PE, phosphatidylethanolamine; PE(O-), alkylphosphatidylethanolamine; PE(P-), alkenylphosphatidylethanolamine; PG, phosphatidylglycerol; PI, phosphatidylinositol; SM, sphingomyelin; Cer, ceramide; HexCer, hexose-ceramides). Statistical analyses were performed by unpaired two-tailed t tests to compare FUS-mutant OPCs and their respective controls (a, b, d, f, g, and i) and two-way ANOVA with the Bonferroni’s multiple comparisons test (c, e, h, and j). Data are represented as mean ± SD. *p < 0.05 **p < 0.01 ****p < 0.0001