Journal: Cell Reports Medicine

Article Title: Metabolic classification suggests the GLUT1/ALDOB/G6PD axis as a therapeutic target in chemotherapy-resistant pancreatic cancer

doi: 10.1016/j.xcrm.2023.101162

Figure Lengend Snippet: Metabolite profiling stratifies PDAC into two subtypes (A) PDAC organoid subgrouping based on widely targeted metabolite abundance. Differential metabolites were identified by the Wilcoxon rank-sum test (p < 0.05, fold change > 1.2). Samples and metabolites are displayed as columns and rows, respectively, and the color of each organoid shows the relative abundance of the metabolites. (B) Relative enrichment of the six metabolic ontology classes in class 1 and class 2 organoids, presented as the difference (class 2 versus class 1) in the ssGSEA score. Positive scores represent terms enriched in metabolites with high intensities in class 2. (C) Fractions of labeled metabolites in glycolysis from [U- 13 C 6 ] glucose in representative organoids of class 1 (n = 6) and class 2 (n = 6). (D) Fractions of labeled metabolites in the TCA cycle from [U- 13 C 6 ] glucose in representative organoids of class 1 (n = 6) and class 2 (n = 6). (E and F) Fractions of labeled metabolites in oxidative PPP (E) and nonoxidative PPP (F) from [U- 13 C 6 ] glucose in representative organoids of class 1 (n = 6) and class 2 (n = 6). (G) Heatmap of widely targeted lipidomics assay showing the abundance of differentially lipid metabolites in class 1 (n = 6) and class 2 (n = 6). (H and I) Class 1 (n = 13) and class 2 (n = 15) organoids were exposed to oligomycin, FCCP, and rotenone/antimycin A to measure the oxygen consumption rate (OCR) by XF Cell Mito Stress Test. (J and K) Dose-response curves and normalized area under the curve (AUC) distribution for TVB-3166 (fatty acid synthase inhibitor) on class 1 and class 2. (L and M) Dose-response curves and normalized AUC distribution for Gboxin (inhibitor of oxidative phosphorylation) on class 1 (n = 13) and class 2 (n = 15). (N) Heatmap of RNA sequencing (RNA-seq) showing the expression of differentially expressed genes (DEGs) in glucomet-PDAC (n = 15) and lipomet-PDAC (n = 13) (p was calculated with limma, p < 0.05 and fold change > 1.5). Representative genes associated with lipid or glucose metabolism are shown in the box on the right. (O) Profile plot and heatmap of assay for transposase-accessible chromatin with sequencing (ATAC-seq) results showing the distribution of differential peaks around the transcriptional start site (TSS) of signature genes in lipomet-PDAC (n = 13) and glucomet-PDAC (n = 15). (P) KEGG pathways enriched in the lipomet subtype (left) and glucomet subtype (right) identified by GSEA. The significance of the difference was determined by Student’s t test (∗p < 0.05; ∗∗p < 0.01) (B). Data are presented as the mean values ± SEMs, and statistical significance was computed by unpaired Student’s t test (C–F). Statistical significance was computed by unpaired Student’s t test (I, K, and M). αKG, α-ketoglutarate; G6P, glucose-6-phosphate; 6PG, 6-phosphate gluconate; F6P, fructose-6-phosphate; R5P, ribose 5-phosphate; S7P, sedoheptulose-7-phosphate; E4P, erythrosine 4-phosphate.

Article Snippet: R5P , MCE , Cat#207671-46-3.

Techniques: Labeling, Phospho-proteomics, RNA Sequencing, Expressing, Sequencing

Journal: Cell Reports Medicine

Article Title: Metabolic classification suggests the GLUT1/ALDOB/G6PD axis as a therapeutic target in chemotherapy-resistant pancreatic cancer

doi: 10.1016/j.xcrm.2023.101162

Figure Lengend Snippet: The GLUT1/ALDOB/G6PD axis contributes to drug resistance (A) Heatmap showing the relative abundance of nucleoside and nucleoside derivatives in glucomet-PDAC (n = 15) and lipomet-PDAC (n = 13). (B) Heatmap showing Pearson correlation coefficients between the RNA expression levels of ALDOB / GLUT1 and nucleoside derivatives. (C and D) DAC-18 (lipomet) and DAC-42 (glucomet) organoids were treated with 5-FU alone or in combination with pyrimidine nucleosides (uridine, cytidine, and thymidine, 240 μM) for 5 days, and cell viability was determined by CellTiter-Glo assays. (E and F) DAC-18 (lipomet) and DAC-42 (glucomet) organoids were treated with PTX alone or in combination with pyrimidine nucleosides (240 μM) for 5 days, and cell viability was determined by CellTiter-Glo assays. (G) Effect of GLUT1 knockdown on 5-FU responsiveness of DAC-34 (glucomet) as determined by CellTiter-Glo assays 120 h after treatment with 5-FU. (H) Effect of ALDOB overexpression on the 5-FU responsiveness of DAC-42 (glucomet) as determined by CellTiter-Glo assays 120 h after treatment with 5-FU. (I) Effect of G6PD knockdown on the 5-FU responsiveness of DAC-42 (glucomet) as determined by CellTiter-Glo assays 120 h after treatment with 5-FU. (J) Effect of ALDOB overexpression on 5-FU responsiveness in the DAC-42 (glucomet) ODX model (n = 6 per group). (K) Effect of pyrimidine nucleotide (240 μM) on 5-FU sensitivity in control and GLUT1 knockdown organoids by CellTiter-Glo assays at 120 h posttreatment. (L) Effect of pyrimidine nucleotide (240 μM) on 5-FU sensitivity in control and ALDOB overexpression organoids by CellTiter-Glo assays at 120 h posttreatment. (M) Effect of R5P (1 mM) on 5-FU sensitivity in control and GLUT1 knockdown organoids by CellTiter-Glo assays at 120 h posttreatment. (N) Effect of R5P (1 mM) on 5-FU sensitivity in control and ALDOB -overexpressing organoids by CellTiter-Glo assays at 120 h posttreatment. (O) Effect of R5P (1 mM) on 5-FU sensitivity in control and G6PD knockdown organoids by CellTiter-Glo assays at 120 h posttreatment. (P and Q) Effect of purine nucleotides (guanosine and adenosine, 200 μM) on 5-FU and GEM sensitivity in control and GLUT1 knockdown organoids by CellTiter-Glo assays at 120 h posttreatment. All dose-responsive curves were performed with 3 technical replicates. Data are presented as the mean values ± SEMs, and statistical significance was computed by unpaired Student’s t test (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001) (C–Q).

Article Snippet: R5P , MCE , Cat#207671-46-3.

Techniques: RNA Expression, Knockdown, Over Expression, Control

Journal: Cell Reports Medicine

Article Title: Metabolic classification suggests the GLUT1/ALDOB/G6PD axis as a therapeutic target in chemotherapy-resistant pancreatic cancer

doi: 10.1016/j.xcrm.2023.101162

Figure Lengend Snippet:

Article Snippet: R5P , MCE , Cat#207671-46-3.

Techniques: Sequencing, Recombinant, SYBR Green Assay, RNA Expression, shRNA, Software