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. 2023 Jul;43(7):838-843.
doi: 10.1002/cac2.12427. Epub 2023 May 4.

DLST mutations in pheochromocytoma and paraganglioma cause proteome hyposuccinylation and metabolic remodeling

Sara Mellid  1 , Fernando García  2 , Luis Javier Leandro-García  1 , Alberto Díaz-Talavera  1   3 , Ángel Mario Martínez-Montes  1 , Eduardo Gil  1 , Bruna Calsina  1 , María Monteagudo  1 , Rocío Letón  1 , Juan María Roldán-Romero  1 , María Santos  1 , Javier Lanillos  1 , Carlos Valdivia  1 , Natalia Martínez-Puente  1   3 , Javier de Nicolás-Hernández  1 , Scherezade Jiménez  4 , Manuel Pérez-Martínez  5 , Emiliano Honrado  6 , Javier Coloma  7 , Ana Cerezo  8 , Clara María Santiveri  9 , Manel Esteller  10   11   12   13 , Ramón Campos-Olivas  9 , Eduardo Caleiras  14 , Cristina Montero-Conde  1 , Cristina Rodríguez-Antona  1 , Javier Muñoz  2   15   16 , Mercedes Robledo  1   3 , Alberto Cascón  1   3
Affiliations

DLST mutations in pheochromocytoma and paraganglioma cause proteome hyposuccinylation and metabolic remodeling

Sara Mellid et al. Cancer Commun (Lond). 2023 Jul.
No abstract available

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIGURE 1
FIGURE 1
PPGL‐causing DLST mutations remodel the cellular succinylome and up‐regulate the hypoxic pathway. (A) Unsupervised clustering of transcriptomic data (z‐scores) for genes expressed differentially in PPGLs (Burnichon et al. 2011) grouped DLST‐mutated tumors (shown in green) with cases featuring SDH/VHL/EPAS1 (in blue, red, and purple, respectively) mutations (Cluster 1) and separately from Cluster 2 PPGLs (tumors carrying mutations in kinase signaling‐related genes; in black), evidencing their pseudohypoxic nature. (B) Hierarchical clustering using DNA methylation data from a list of probes found differentially methylated in SDHB‐ or FH‐mutated tumors showing a CpG island methylator phenotype (Ricketts CJ et al. PLoS One. 2022;17(12):e0278108), separated DLST‐mutated tumors (green) from hypermethylated tumors carrying mutations in SDH genes (blue). (C) Structure of DLST's homooligomeric 24‐mer. Three DLST monomers are highlighted in dark blue, yellow and purple. The upper panels show the structure of DLST WT, while the bottom left and bottom right panels show the p.G374E and p.Y422C mutants, respectively. SucCoa denotes Succinyl‐CoA (only the succinyl group is shown). (D‐G) Volcano plots showing the number of differentially succinylated sites (Log2FC < ‐1 or > 1, P < 0.05) in DLST KO (D), DLST H424A (E), DLST G374E (F), and DLST Y422C (G) cells when compared to DLST WT cells. Blue, red and grey dots represent differentially hyposuccinylated sites, differentially hypersuccinylated sites, and non‐significantly succinylated sites and/or with a Log2FC between ‐1 and 1, respectively. (H) Representation of sites exhibiting significant differential succinylation (colored circles) in TCAc enzymes (left panel) and the Glycolysis/Gluconeogenesis pathway (right panel) when comparing DLST G374E and WT DLST cells. Both pathways exhibit global hyposuccinylation (blue color). (I) Anti‐succinyllysine immunohistochemistry images of PPGLs harboring the DLST mutations p.Y422C (left top) and p.G374E (left bottom) and known mutations in other susceptibility genes as controls (right). The granular pattern and perinuclear staining observed in the images on the right suggest mitochondrial staining, whereas cells from DLST‐mutated PPGLs exhibit more homogeneous cytoplasmic staining. The tumor harboring the p.Y422C DLST mutation (top left) has an intermediate pattern with fewer granules and some perinuclear staining, in accordance with proteomic succinylation data obtained using our cell model. (J) Immunofluorescence analysis of the different cell lines showing that DLST (green) colocalizes with the Mitotracker dye (red, MITO in the figure) and is thus predominantly localized in the mitochondria regardless of its mutations in all cells other than DLST KO cells lacking the DLST protein. Nuclei are stained in blue with DAPI. (K) Immunoblotting assay showing that DLST is present in the cytosol and nucleus compartments as well as the mitochondria‐enriched fraction (named mitochondria in the figure) regardless of PPGL‐causing mutations. OGDH levels are significantly reduced in DLST KO cells lacking DLST. GAPDH and TBP (nucleus) were used as loading controls, and COXIV as a mitochondrial marker. (L) Representation of normalized enrichment scores (NES) relative to DLST WT cells for the GSEA Hallmark gene sets of Hypoxia, Glycolysis and Oxidative phosphorylation in cells harboring DLST alterations (from left to right: DLST H424A, DLST G374E, DLST Y422C, DLST KO and LCL‐G374E). Abbreviations: PPGL, pheochromocytoma and paraganglioma; SucCoA, succinyl coenzyme A; TCAc, tricarboxylic acid cycle; DAPI, 4’,6‐diamidino‐2‐phenylindole, dihydrochloride; NES, normalized enrichment score; GSEA, gene set enrichment analysis; LCL, lymphoblastoid cell line; Succ‐K IHC, succinylated lysine immunohistochemistry; Log2FC, Log2 fold change.
See this image and copyright information in PMC

References

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