Integrated lipidomics and proteomics reveal cardiolipin alterations, upregulation of HADHA and long chain fatty acids in pancreatic cancer stem cells.
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ABSTRACT:
SUBMITTER: Di Carlo C
PROVIDER: S-EPMC8225828 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Pancreatic cancer stem cells (PCSCs) characterize the pancreatic ductal adenocarcinomas (PDAC) and play a key role in driving their poor outcomes and resistance to targeted therapies. Although advanced proteomics and lipidomics technology facilitates quantitation of intracellular proteins and lipids, little is known about the regulation of signalling and metabolic pathways in PCSCs. Here we shown that PCSCs, derived from different PDAC cell lines, have specific and common proteome and lipidome modulations. PCSCs displayed down-regulation of L-lactate dehydrogenase A chain (LDHA) and, more interestingly, upregulation of trifunctional enzyme subunit alpha (HADHA) which is involved in cardiolipin remodelling. The upregulated proteins of PCSCs were mainly involved in fatty acid (FA) elongation and biosynthesis of unsaturated FAs. Accordingly, lipidomics revealed a general increase of FAs 18:1, 20:2, 20:3, and 20:4, which are products of the catalytic activity of fatty acid elongase-5 (ELOVL5) predicted as common active gene of PCSCs. Moreover, in accordance with HADHA upregulation, lipidomics also showed the induction in PCSCs of molecular species of cardiolipin with mixed incorporation of 16:0, 18:1, and 18:2 acyl chains. Our data indicate a crucial role of fatty acid elongation and cardiolipin remodelling in PCSCs, which could therefore represent attractive targets to improve the therapeutic treatments of PDAC
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Project description:Mitochondrial trifunctional protein deficiency, due to mutations in hydratase subunit A (HADHA), results in sudden infant death syndrome with no cure. To reveal the disease etiology, we generated stem cell-derived cardiomyocytes from HADHA-deficient hiPSCs and accelerated their maturation via an engineered microRNA maturation cocktail that upregulated the epigenetic regulator, HOPX. Here we report, matured HADHA mutant cardiomyocytes treated with an endogenous mixture of fatty acids manifest the disease phenotype: defective calcium dynamics and repolarization kinetics which results in a pro-arrhythmic state. Single cell RNA-seq reveals a cardiomyocyte developmental intermediate, based on metabolic gene expression. This intermediate gives rise to mature-like cardiomyocytes in control cells but, mutant cells transition to a pathological state with reduced fatty acid beta-oxidation, reduced mitochondrial proton gradient, disrupted cristae structure and defective cardiolipin remodeling. This study reveals that HADHA (tri-functional protein alpha), a monolysocardiolipin acyltransferase-like enzyme, is required for fatty acid beta-oxidation and cardiolipin remodeling, essential for functional mitochondria in human cardiomyocytes.
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| S-EPMC5831854 | biostudies-other
Project description:Mitochondrial trifunctional protein deficiency, due to mutations in hydratase subunit A (HADHA), results in sudden infant death syndrome with no cure. To reveal the disease etiology, we generated stem cell-derived cardiomyocytes from HADHA-deficient hiPSCs and accelerated their maturation via an engineered microRNA maturation cocktail that upregulated the epigenetic regulator, HOPX. Here we report, matured HADHA mutant cardiomyocytes treated with an endogenous mixture of fatty acids manifest the disease phenotype: defective calcium dynamics and repolarization kinetics which results in a pro-arrhythmic state. Single cell RNA-seq reveals a cardiomyocyte developmental intermediate, based on metabolic gene expression. This intermediate gives rise to mature-like cardiomyocytes in control cells but, mutant cells transition to a pathological state with reduced fatty acid beta-oxidation, reduced mitochondrial proton gradient, disrupted cristae structure and defective cardiolipin remodeling. This study reveals that HADHA (tri-functional protein alpha), a monolysocardiolipin acyltransferase-like enzyme, is required for fatty acid beta-oxidation and cardiolipin remodeling, essential for functional mitochondria in human cardiomyocytes.
2019-08-25 | GSE135595 | GEO