Proteomic and Molecular Assessment of the Common Saudi Variant in ACADVL Gene Through Mesenchymal Stem Cells

Very-long-chain acyl-coenzyme A dehydrogenase (VLCAD) is a coenzyme encoded by ACADVL that converts very-long-chain fatty acids into energy. This process is disrupted by c.65C > A; p.Ser22∗ mutation. To clarify mechanisms by which this mutation leads to VLCAD deficiency, we evaluated differences in molecular and cellular functions between mesenchymal stem cells with normal and mutant VLCAD. Saudi Arabia have a high incidence of this form of mutation. Stem cells with mutant VLCAD were isolated from skin of two patients. Metabolic activity and proliferation were evaluated. The Same evaluation was repeated on normal stem cells introduced with same mutation by CRISPR. Mitochondrial depiction was done by electron microscope and proteomic analysis was done on patients’ cells. Metabolic activity and proliferation were significantly lower in patients’ cells. Introducing the same mutation into normal stem cells resulted in the same defects. We detected mitochondrial abnormalities by electron microscopy in addition to poor wound healing and migration processes in mutant cells. Furthermore, in a proteomic analysis, we identified several upregulated or downregulated proteins related to hypoglycemia, liver disorder, and cardiac and muscle involvement. We concluded experimental assays of mutant ACADVL (c.65C > A; p.Ser22∗) contribute to severe neonatal disorders with hypoglycemia, liver disorder, and cardiac and muscle involvement.

[1]  Wei Cheng,et al.  Very Long-Chain Acyl-Coenzyme A Dehydrogenase Deficiency , 2014 .

[2]  Yanling Yang,et al.  One Potential Hotspot ACADVL Mutation in Chinese Patients with Very-long-chain Acyl-coenzyme A Dehydrogenase Deficiency. , 2019, Clinica chimica acta; international journal of clinical chemistry.

[3]  Bahauddeen M. Alrfaei,et al.  The SORCS3 gene is mutated in brothers with infantile spasms and intellectual disability. , 2018, Discovery medicine.

[4]  Dong Hwan Lee,et al.  Diversity in the incidence and spectrum of organic acidemias, fatty acid oxidation disorders, and amino acid disorders in Asian countries: Selective screening vs. expanded newborn screening , 2018, Molecular genetics and metabolism reports.

[5]  S. Ruet,et al.  Development of a Tandem Mass Spectrometry Method for Rapid Measurement of Medium- and Very-Long-Chain Acyl-CoA Dehydrogenase Activity in Fibroblasts. , 2016, JIMD reports.

[6]  S. Tucci,et al.  Triheptanoin: long-term effects in the very long-chain acyl-CoA dehydrogenase-deficient mouse[S] , 2016, Journal of Lipid Research.

[7]  M. Alfadhel,et al.  Thirteen year retrospective review of the spectrum of inborn errors of metabolism presenting in a tertiary center in Saudi Arabia , 2016, Orphanet Journal of Rare Diseases.

[8]  Yun Zhang,et al.  Correction: Mouse SIRT3 Attenuates Hypertrophy-Related Lipid Accumulation in the Heart through the Deacetylation of LCAD , 2016, PloS one.

[9]  W. Craigen,et al.  Recurrent ACADVL molecular findings in individuals with a positive newborn screen for very long chain acyl-coA dehydrogenase (VLCAD) deficiency in the United States. , 2015, Molecular genetics and metabolism.

[10]  J. Merritt,et al.  Orphan drugs in development for long-chain fatty acid oxidation disorders: challenges and progress , 2015 .

[11]  Matthew J. Rardin,et al.  SIRT3 and SIRT5 Regulate the Enzyme Activity and Cardiolipin Binding of Very Long-Chain Acyl-CoA Dehydrogenase , 2015, PloS one.

[12]  A. Kaye,et al.  Rhabdomyolysis: pathogenesis, diagnosis, and treatment. , 2015, The Ochsner journal.

[13]  R. Singer,et al.  The translation elongation factor eEF1A1 couples transcription to translation during heat shock response , 2014, eLife.

[14]  M. Young,et al.  Use of phage display to identify novel mineralocorticoid receptor-interacting proteins. , 2014, Molecular endocrinology.

[15]  Sander M Houten,et al.  Mitochondrial protein acetylation is driven by acetyl-CoA from fatty acid oxidation. , 2014, Human molecular genetics.

[16]  A. Masood,et al.  Proteomic analysis of mature adipo cytes from obese patients in relation to aging , 2013, Experimental Gerontology.

[17]  B. Schüle,et al.  Skin punch biopsy explant culture for derivation of primary human fibroblasts. , 2013, Journal of visualized experiments : JoVE.

[18]  Gary L Hoffman,et al.  Clinical validation of cutoff target ranges in newborn screening of metabolic disorders by tandem mass spectrometry: A worldwide collaborative project , 2011, Genetics in Medicine.

[19]  Bridget Wilcken,et al.  Fatty acid oxidation disorders: outcome and long-term prognosis , 2010, Journal of Inherited Metabolic Disease.

[20]  M. Baumgartner,et al.  Management and outcome in 75 individuals with long-chain fatty acid oxidation defects: results from a workshop , 2009, Journal of Inherited Metabolic Disease.

[21]  Yudong Wang,et al.  Structural Basis for Substrate Fatty Acyl Chain Specificity , 2008, Journal of Biological Chemistry.

[22]  B. V. van Engelen,et al.  Rhabdomyolysis caused by an inherited metabolic disease: very long-chain acyl-CoA dehydrogenase deficiency. , 2006, The American journal of medicine.

[23]  D. Vertommen,et al.  Myocardial Ischemia and Increased Heart Work Modulate the Phosphorylation State of Eukaryotic Elongation Factor-2* , 2003, Journal of Biological Chemistry.

[24]  Guillermo Oliver,et al.  Hepatocyte migration during liver development requires Prox1 , 2000, Nature Genetics.

[25]  William,et al.  Purification of human very-long-chain acyl-coenzyme A dehydrogenase and characterization of its deficiency in seven patients. , 1995, The Journal of clinical investigation.

[26]  C. Stanley,et al.  Long-Chain Acyl Coenzyme A Dehydrogenase Deficiency: An Inherited Cause of Nonketotic Hypoglycemia , 1985, Pediatric Research.

[27]  Walter S. Krawczyk,et al.  A PATTERN OF EPIDERMAL CELL MIGRATION DURING WOUND HEALING , 1971, The Journal of cell biology.