Tissue Proteome of 2-Hydroxyacyl-CoA Lyase Deficient Mice Reveals Peroxisome Proliferation and Activation of ω-Oxidation

• Published in: International journal of molecular sciences Vol. 23; no. 2; p. 987
• Main Authors: Khalil, Youssef, Carrino, Sara, Lin, Fujun, Ferlin, Anna, Lad, Heena V, Mazzacuva, Francesca, Falcone, Sara, Rivers, Natalie, Banks, Gareth, Concas, Danilo, Aguilar, Carlos, Haynes, Andrew R, Blease, Andy, Nicol, Thomas, Al-Shawi, Raya, Heywood, Wendy, Potter, Paul, Mills, Kevin, Gale, Daniel P, Clayton, Peter T
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 17.01.2022; MDPI
• Subjects: Animals; Arachidonic acid; Ataxia; Brain - metabolism; Carbon; Carbon-Carbon Lyases - genetics; Cytochrome P450 Family 2 - metabolism; Cytochrome P450 Family 4 - metabolism; Diet; Fatty acids; Fatty Acids - metabolism; Female; Gas chromatography; Gene Knockout Techniques; Genotype & phenotype; Hydroxylase; isoprenoids; Kidney - metabolism; Kidneys; Lipidomics - methods; Liquid chromatography; Liver; Liver - metabolism; Male; Mass spectrometry; Mass spectroscopy; Metabolomics; Mice; Oxidation; Oxidation-Reduction; Peroxisome proliferator-activated receptors; peroxisomes; Peroxisomes - metabolism; Phenotypes; Phytanic Acid - analogs & derivatives; Phytanic Acid - metabolism; Phytol; Phytol - administration & dosage; Phytol - pharmacology; PPARs; Proteins; Proteomes; Proteomics; Proteomics - methods; Refsum's disease; Signal transduction; Spectroscopy; Trypsin; United Kingdom > UK; United States > US; peroxisomes; proteomics; isoprenoids; liver; PPARs
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23020987

Peroxisomal fatty acid α-oxidation is an essential pathway for the degradation of β-carbon methylated fatty acids such as phytanic acid. One enzyme in this pathway is 2-hydroxyacyl CoA lyase (HACL1), which is responsible for the cleavage of 2-hydroxyphytanoyl-CoA into pristanal and formyl-CoA. Hacl1 deficient mice do not present with a severe phenotype, unlike mice deficient in other α-oxidation enzymes such as phytanoyl-CoA hydroxylase deficiency (Refsum disease) in which neuropathy and ataxia are present. Tissues from wild-type and mice fed a high phytol diet were obtained for proteomic and lipidomic analysis. There was no phenotype observed in these mice. Liver, brain, and kidney tissues underwent trypsin digestion for untargeted proteomic liquid chromatography-mass spectrometry analysis, while liver tissues also underwent fatty acid hydrolysis, extraction, and derivatisation for fatty acid gas chromatography-mass spectrometry analysis. The liver fatty acid profile demonstrated an accumulation of phytanic and 2-hydroxyphytanic acid in the liver and significant decrease in heptadecanoic acid. The liver proteome showed a significant decrease in the abundance of Hacl1 and a significant increase in the abundance of proteins involved in PPAR signalling, peroxisome proliferation, and omega oxidation, particularly Cyp4a10 and Cyp4a14. In addition, the pathway associated with arachidonic acid metabolism was affected; Cyp2c55 was upregulated and Cyp4f14 and Cyp2b9 were downregulated. The kidney proteome revealed fewer significantly upregulated peroxisomal proteins and the brain proteome was not significantly different in mice. This study demonstrates the powerful insight brought by proteomic and metabolomic profiling of mice in better understanding disease mechanism in fatty acid α-oxidation disorders.