ZFIN Image: Jamalpoor et al., 2021, Figure 2

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Figure Caption

Figure 2 Metabolomic and proteomic profiling reveal αKG accumulation in cystinosis

Principal component analysis (PCA) of control (CTNS^WT), CRISPR‐generated cystinotic (CTNS^−/−) and patient‐derived cystinotic (CTNS^Patient) cells based on the metabolites measured. Each dot represents one sample, and the dots of the same colour are biological replicates (n = 3).

Heat map analysis of top 25 metabolites distinctively expressed in control and CTNS^−/− cells. Metabolites significantly decreased (P < 0.05) were displayed in green, while metabolites significantly increased (P < 0.05) were displayed in red (n = 3).

Global test pathway enrichment analysis of the intracellular metabolic interactions distinctively affected in CTNS^−/− cells compared to healthy control cells (n = 3). Larger circles further from the y‐axis and orange‐red colour show higher impact of pathway.

List of metabolites that were shared and significantly altered in both the CTNS^−/− and CTNS^Patient cells compared to control cells (n = 6).

Plasma levels of αKG (µM) in healthy individuals (n = 4) and cystinotic patients (n = 6). All cystinotic patients were on cysteamine treatment at the time of blood sampling. For this experiment, non‐parametric Mann–Whitney t‐test was used to demonstrate the significance.

Principal component analysis (PCA) of the measured proteins in CTNS^WT, CTNS^−/− and CTNS^Patient (n = 3).

Volcano plot illustrates differentially abundant proteins (n = 3). The ‐log₁₀ (Benjamini–Hochberg corrected P‐value) is plotted against the log₂ (fold change: CTNS^WT/CTNS^−/−).

Gene ontology (GO) analysis illustrates classes of proteins differing between CTNS^WT and CTNS^−/− cells (n = 3).

List of proteins that were significantly upregulated and downregulated in CTNS^−/− cells compared to control cells (n = 3). AKGDH; Alpha‐ketoglutarate dehydrogenase, LIPA; Lysosomal acid lipase, ACP2; Lysosomal acid phosphatase, CTSS; Cathepsin S, CTSC; Dipeptidyl peptidase, IGF2R; Cation‐independent mannose‐6‐phosphate receptor, SORT1; Sortilin, CAT; Catalase, CTSA; Lysosomal protective protein and SOD; Superoxide dismutase.

Data information: Data are expressed as mean ± SEM. *P‐values < 0.05 were considered to be significant. One‐way ANOVA with Dunnett’s correction (A, B, C, D, E, F, G, H and I). Exact P‐values and statistical tests are listed in Appendix Table S1.

Acknowledgments

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