Fig. 6: Dual inhibition of FAO and AURKA elicits a synergistic reduction in cellular viability of GBM cells.

For a, b parental or chronically alisertib treated GBM22 cells were subjected to transcriptomic analysis and followed by GSEA. Shown are a graphical plot of NES vs FDR-q value of microarray data in (a) and an enrichment plot of Go_Regulation_of_Fatty_Acid_Oxidation in (b). NES: normalized enrichment score. FDR: false discovery rate. c Real-time PCR analysis of genes related to fatty acid oxidation (mRNA levels) of parental or chronically alisertib treated GBM22 cells (n = 8 in GBM22 and n = 4 in GBM22AR independent samples). FC: fold change. d Non-polar metabolite analysis of parental or chronically alisertib treated GBM22 cells. A heatmap of parental or chronically alisertib treated GBM22 cells is shown. AcCa: acyl carnitine; Cer: ceramides; CerG1-3: neutral glycosphingolipids. CerP: phosphosphingolipids; ChE: cholesterol Ester; DG: diglyceride; LPC: lysophosphatidylcholine; LPE lysophosphatidylethanolamine; LPI: lysophosphatidylinositol; LPS: lysophosphatidylserine; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PG: phosphatidylglycerol; PI: phosphatidylinositol; PS: phosphatidylserine; SM: sphingomyelin; So: sphingosine; TG: triglyceride. For e, f acyl-carnitine and lysophosphatidyl levels in the non-polar metabolite analysis of parental or chronically alisertib treated GBM22 cells (n = 5 independent samples). For g, h Parental or chronically alisertib treated GBM22 and SF188 cells were treated with alisertib, etomoxir, or the combination of both for 72 h, and cellular viability was analyzed. Isobolograms are shown in (g) and quantification of cell viability is shown in (h) (n = 4 independent samples) (**p = 0.0047, ***p = 0.0002, ****p < 0.0001). i Parental or chronically alisertib treated GBM22 cells were treated with alisertib, etomoxir, or the combination of both for 24 h and analyzed for oxygen consumption rate (OCR) on a Seahorse XFe24 device. The graph (right panel) shows the OCR level (n = 5 independent samples) (*p = 0.0195, ****p < 0.0001). j SF188 cells were transfected with non-targeting siNT or siAURKA (single or pool), treated with etomoxir for 72 h, and cellular viability was analyzed (n = 4 independent samples) (siAURKA-2: *p = 0.0181, siAURKA-4: *p = 0.1015). For k, l SF188 cells were transfected with non-targeting siNT or CPT1A specific siRNA (siCPT1A), treated with alisertib for 72 h, and cellular viability was analyzed (n = 7 in siNT + DMSO, n = 8 in siCPT1A + DMSO, n = 4 in siNT + Ali 20 nM and siCPT1A + Ali 20 nM independent samples) (*p = 0.0495, ***p = 0.0009). Protein capillary electrophoresis confirms the silencing of CPT1A is shown in l. Vinculin is used as a loading control. Statistical significance was assessed by a two-tailed student’s t-test in (j, k) or ANOVA with Dunnett’s multiple comparison test in (h, i). Data are shown as mean ± SD in (c, e, h−k). Source data are provided as a Source Data file.