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1.
Mol Genet Metab ; 116(4): 281-8, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26549575

ABSTRACT

Coenzyme A (CoA) is a ubiquitous cofactor involved in numerous essential biochemical transformations, and along with its thioesters is a key regulator of intermediary metabolism. Pantothenate (vitamin B5) phosphorylation by pantothenate kinase (PanK) is thought to control the rate of CoA production. Pantothenate kinase associated neurodegeneration is a hereditary disease that arises from mutations that inactivate the human PANK2 gene. Aryl phosphoramidate phosphopantothenate derivatives were prepared to test the feasibility of using phosphopantothenate replacement therapy to bypass the genetic deficiency in the Pank1(-/-) mouse model. The efficacies of candidate compounds were first compared by measuring the ability to increase CoA levels in Pank1(-/-) mouse embryo fibroblasts. Administration of selected candidate compounds to Pank1(-/-) mice corrected their deficiency in hepatic CoA. The PanK bypass was confirmed by the incorporation of intact phosphopantothenate into CoA using triple-isotopically labeled compound. These results provide strong support for PanK as a master regulator of intracellular CoA and illustrate the feasibility of employing PanK bypass therapy to restore CoA levels in genetically deficient mice.


Subject(s)
Amides/pharmacology , Coenzyme A/biosynthesis , Liver/drug effects , Pantothenate Kinase-Associated Neurodegeneration/diet therapy , Pantothenic Acid/analogs & derivatives , Phosphoric Acids/pharmacology , Phosphotransferases (Alcohol Group Acceptor)/deficiency , Administration, Oral , Amides/chemical synthesis , Animals , Coenzyme A/deficiency , Coenzyme A/genetics , Disease Models, Animal , Embryo, Mammalian , Female , Fibroblasts/drug effects , Fibroblasts/enzymology , Fibroblasts/pathology , Gene Expression , Humans , Liver/enzymology , Liver/pathology , Male , Mice , Mice, Knockout , Pantothenate Kinase-Associated Neurodegeneration/enzymology , Pantothenate Kinase-Associated Neurodegeneration/genetics , Pantothenate Kinase-Associated Neurodegeneration/pathology , Pantothenic Acid/chemical synthesis , Pantothenic Acid/pharmacology , Phosphoric Acids/chemical synthesis , Phosphotransferases (Alcohol Group Acceptor)/genetics , Primary Cell Culture
2.
Mol Genet Metab ; 105(3): 463-71, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22221393

ABSTRACT

Pantothenate kinase-associated neurodegeneration (PKAN) is a rare, inborn error of metabolism characterized by iron accumulation in the basal ganglia and by the presence of dystonia, dysarthria, and retinal degeneration. Mutations in pantothenate kinase 2 (PANK2), the rate-limiting enzyme in mitochondrial coenzyme A biosynthesis, represent the most common genetic cause of this disorder. How mutations in this core metabolic enzyme give rise to such a broad clinical spectrum of pathology remains a mystery. To systematically explore its pathogenesis, we performed global metabolic profiling on plasma from a cohort of 14 genetically defined patients and 18 controls. Notably, lactate is elevated in PKAN patients, suggesting dysfunctional mitochondrial metabolism. As predicted, but never previously reported, pantothenate levels are higher in patients with premature stop mutations in PANK2. Global metabolic profiling and follow-up studies in patient-derived fibroblasts also reveal defects in bile acid conjugation and lipid metabolism, pathways that require coenzyme A. These findings raise a novel therapeutic hypothesis, namely, that dietary fats and bile acid supplements may hold potential as disease-modifying interventions. Our study illustrates the value of metabolic profiling as a tool for systematically exploring the biochemical basis of inherited metabolic diseases.


Subject(s)
Coenzyme A/deficiency , Mitochondria/enzymology , Neuroaxonal Dystrophies/metabolism , Pantothenate Kinase-Associated Neurodegeneration/metabolism , Phosphotransferases (Alcohol Group Acceptor)/genetics , Adolescent , Adult , Bile Acids and Salts/metabolism , Child , Child, Preschool , Codon, Nonsense , Coenzyme A/biosynthesis , Coenzyme A/genetics , Cohort Studies , Female , Humans , Iron Metabolism Disorders , Lactic Acid/blood , Lipid Metabolism/genetics , Lipid Metabolism Disorders/genetics , Lipid Metabolism Disorders/metabolism , Male , Metabolome , Mitochondria/genetics , Mitochondria/metabolism , Mitochondria/pathology , Neuroaxonal Dystrophies/diagnosis , Neuroaxonal Dystrophies/enzymology , Pantothenate Kinase-Associated Neurodegeneration/enzymology , Pantothenate Kinase-Associated Neurodegeneration/genetics , Pantothenic Acid/blood , Sphingomyelins/blood , Young Adult
3.
Biochim Biophys Acta Mol Basis Dis ; 1866(5): 165663, 2020 05 01.
Article in English | MEDLINE | ID: mdl-31918006

ABSTRACT

Pantothenate kinase (PanK) is the first enzyme in the coenzyme A (CoA) biosynthetic pathway. The differential expression of the four-active mammalian PanK isoforms regulates CoA levels in different tissues and PANK2 mutations lead to Pantothenate Kinase Associated Neurodegeneration (PKAN). The molecular mechanisms that potentially underlie PKAN pathophysiology are investigated in a mouse model of CoA deficiency in the central nervous system (CNS). Both PanK1 and PanK2 contribute to brain CoA levels in mice and so a mouse model with a systemic deletion of Pank1 together with neuronal deletion of Pank2 was generated. Neuronal Pank2 expression in double knockout mice decreased starting at P9-11 triggering a significant brain CoA deficiency. The depressed brain CoA in the mice correlates with abnormal forelimb flexing and weakness that, in turn, contributes to reduced locomotion and abnormal gait. Biochemical analysis reveals a reduction in short-chain acyl-CoAs, including acetyl-CoA and succinyl-CoA. Comparative gene expression analysis reveals that the CoA deficiency in brain is associated with a large elevation of Hif3a transcript expression and significant reduction of gene transcripts in heme and hemoglobin synthesis. Reduction of brain heme levels is associated with the CoA deficiency. The data suggest a response to oxygen/glucose deprivation and indicate a disruption of oxidative metabolism arising from a CoA deficiency in the CNS.


Subject(s)
Brain Chemistry/genetics , Brain/pathology , Coenzyme A/deficiency , Pantothenate Kinase-Associated Neurodegeneration/pathology , Phosphotransferases (Alcohol Group Acceptor)/deficiency , Animals , Apoptosis Regulatory Proteins/metabolism , Brain/cytology , Coenzyme A/analysis , Coenzyme A/biosynthesis , Disease Models, Animal , Female , Gene Expression Profiling , Gene Expression Regulation/genetics , Heme/analysis , Heme/metabolism , Hemoglobins/analysis , Hemoglobins/metabolism , Humans , Male , Mice , Mice, Knockout , Neurons/metabolism , Neurons/pathology , Oxidation-Reduction , Pantothenate Kinase-Associated Neurodegeneration/genetics , Phosphotransferases (Alcohol Group Acceptor)/genetics , Repressor Proteins/metabolism
4.
Nat Commun ; 9(1): 4399, 2018 10 23.
Article in English | MEDLINE | ID: mdl-30352999

ABSTRACT

Pantothenate kinase (PANK) is a metabolic enzyme that regulates cellular coenzyme A (CoA) levels. There are three human PANK genes, and inactivating mutations in PANK2 lead to pantothenate kinase associated neurodegeneration (PKAN). Here we performed a library screen followed by chemical optimization to produce PZ-2891, an allosteric PANK activator that crosses the blood brain barrier. PZ-2891 occupies the pantothenate pocket and engages the dimer interface to form a PANK•ATP•Mg2+•PZ-2891 complex. The binding of PZ-2891 to one protomer locks the opposite protomer in a catalytically active conformation that is refractory to acetyl-CoA inhibition. Oral administration of PZ-2891 increases CoA levels in mouse liver and brain. A knockout mouse model of brain CoA deficiency exhibited weight loss, severe locomotor impairment and early death. Knockout mice on PZ-2891 therapy gain weight, and have improved locomotor activity and life span establishing pantazines as novel therapeutics for the treatment of PKAN.


Subject(s)
Pantothenate Kinase-Associated Neurodegeneration/therapy , Phosphotransferases (Alcohol Group Acceptor)/genetics , Adenosine Triphosphate/metabolism , Allosteric Regulation , Animals , Cells, Cultured , Coenzyme A/deficiency , Coenzyme A/metabolism , Disease Models, Animal , Enzyme Stability , Female , Ligands , Magnesium/metabolism , Male , Mice, Knockout , Neurons/metabolism , Organ Specificity , Pantothenate Kinase-Associated Neurodegeneration/pathology , Protein Conformation , Protein Multimerization
5.
J Nutr Sci Vitaminol (Tokyo) ; 35(1): 11-23, 1989 Feb.
Article in English | MEDLINE | ID: mdl-2738713

ABSTRACT

When 400 mg/rat/day of secondary autoxidation products of linoleic acid was orally administered 3 times to rats, they died at 30-40 h after the third dose. To search the markers of the toxicity of secondary products in vivo, the rats were killed at 24h after the third dose, and conditions of their digestive tracts and liver were analyzed. In the stomach, macroscopically, inflation, retention of undigested food, and edema were seen. Slight congestions were detected in the small intestines. It was considered that these injuries led to reduction in food consumption and then depression of the growth, but did not lead to the death of the animals. The lipid peroxide levels in the liver and the activities of its detoxifying enzymes were increased as compared to those in the control groups. The hepatic lipid contents and unsaturated fatty acid compositions were also not changed. The endogenous lipid peroxidation, therefore, did not give the rats a severe stress. The activities of hepatic acetyl-CoA carboxylase and carnitine palmitoyltransferase were 20 and 35% lower than those of control, respectively. The levels of CoASH, acetyl-CoA, and long-chain acyl-CoA were 1/9, 1/2, and 1/4 of those in control, respectively. Thus, one of the markers of the toxicity of secondary products was the depletion of hepatic CoA derivatives. In rat, bio-energy was reduced by the decrease in the intestinal absorption of nutrients, and the depletion of hepatic CoA derivatives also failed to supply energy with beta-oxidation.


Subject(s)
Coenzyme A/deficiency , Linoleic Acids/toxicity , Liver/metabolism , Animals , Body Weight/drug effects , Digestive System/drug effects , Linoleic Acids/administration & dosage , Linoleic Acids/metabolism , Lipid Peroxides/analysis , Liver/drug effects , Male , Organ Size/drug effects , Oxidation-Reduction , Rats , Rats, Inbred Strains , Vitamin E/blood
6.
Mol Cell Biochem ; 175(1-2): 37-42, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9350031

ABSTRACT

Hepatic coenzyme A (CoA) plays an important role in cellular lipid metabolism. Because mitochondria and peroxisomes represent the two major subcellular sites of lipid metabolism, the present study was designed to investigate the specific impact of hepatic CoA deficiency on peroxisomal as well as mitochondrial beta-oxidation of fatty acids. CoA deficiency (47% decrease in free CoA and 23% decrease in total CoA) was produced by maintaining weanling male Sprague-Dawley rats on a semipurified diet deficient in pantothenic acid (the precursor of CoA) for 5 weeks. Hepatic mitochondrial fatty acid oxidation of short-chain and long-chain fatty acids were not significantly different between control and CoA-deficient rats. Conversely, peroxisomal beta-oxidation was significantly diminished (38% inhibition) in livers of CoA-deficient rats compared to control animals. Peroxisomal beta-oxidation was restored to normal levels when hepatic CoA was replenished. It is postulated that since the role of hepatic mitochondrial beta-oxidation is energy production while peroxisomal beta-oxidation acts mainly as a detoxification system, the mitochondrial pathway of beta-oxidation is spared at the expense of the peroxisomal pathway when liver CoA plummets. The present study may offer an animal model to investigate mechanisms involved in peroxisomal diseases.


Subject(s)
Coenzyme A/physiology , Fatty Acids/metabolism , Liver/enzymology , Microbodies/metabolism , Mitochondria, Liver/metabolism , Animals , Coenzyme A/deficiency , Liver/metabolism , Male , Oxidation-Reduction , Pantothenic Acid/deficiency , Pantothenic Acid/metabolism , Rats , Rats, Sprague-Dawley
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