The Experts below are selected from a list of 153 Experts worldwide ranked by ideXlab platform
Stephen D. Cederbaum - One of the best experts on this subject based on the ideXlab platform.
-
lipid nanoparticle targeted mrna therapy as a treatment for the inherited metabolic liver disorder arginase deficiency
Proceedings of the National Academy of Sciences of the United States of America, 2019Co-Authors: Brian Truong, Stephen D. Cederbaum, Gabriella Allegri, Kristine E Burke, Johannes Haberle, Paolo Martini, Gerald S. LipshutzAbstract:Arginase deficiency is caused by biallelic mutations in arginase 1 (ARG1), the final step of the urea cycle, and results biochemically in Hyperargininemia and the presence of guanidino compounds, while it is clinically notable for developmental delays, spastic diplegia, psychomotor function loss, and (uncommonly) death. There is currently no completely effective medical treatment available. While preclinical strategies have been demonstrated, disadvantages with viral-based episomal-expressing gene therapy vectors include the risk of insertional mutagenesis and limited efficacy due to hepatocellular division. Recent advances in messenger RNA (mRNA) codon optimization, synthesis, and encapsulation within biodegradable liver-targeted lipid nanoparticles (LNPs) have potentially enabled a new generation of safer, albeit temporary, treatments to restore liver metabolic function in patients with urea cycle disorders, including ARG1 deficiency. In this study, we applied such technologies to successfully treat an ARG1-deficient murine model. Mice were administered LNPs encapsulating human codon-optimized ARG1 mRNA every 3 d. Mice demonstrated 100% survival with no signs of hyperammonemia or weight loss to beyond 11 wk, compared with controls that perished by day 22. Plasma ammonia, arginine, and glutamine demonstrated good control without elevation of guanidinoacetic acid, a guanidino compound. Evidence of urea cycle activity restoration was demonstrated by the ability to fully metabolize an ammonium challenge and by achieving near-normal ureagenesis; liver arginase activity achieved 54% of wild type. Biochemical and microscopic data showed no evidence of hepatotoxicity. These results suggest that delivery of ARG1 mRNA by liver-targeted nanoparticles may be a viable gene-based therapeutic for the treatment of arginase deficiency.
-
Newborn screening for Hyperargininemia due to arginase 1 deficiency
Molecular genetics and metabolism, 2017Co-Authors: Bradford L. Therrell, Robert Currier, David Lapidus, Meredith Grimm, Stephen D. CederbaumAbstract:Abstract Hyperargininemia caused by Arginase 1 deficiency is a rare disorder of the urea cycle that can be diagnosed by elevation of arginine in newborn screening blood spots when analyzed by tandem mass spectrometry. Hyperargininemia is currently included as a secondary target on the U.S. Recommended Uniform Screening Panel, which directly influences state-based newborn screening. Because of the apparent low disease frequency and lack of case detection and treatment data, detailed attention has not been given to a model newborn screening algorithm including appropriate analytical cutoff values for disease indicators. In this paper we assess the frequency of Hyperargininemia in the U.S. identified by newborn screening to date and document the current status and variability of Hyperargininemia newborn screening across U.S. newborn screening programs. We also review other data that support improved screening efficacy by utilizing the arginine/ornithine ratio and other amino acid ratios as discriminators in the screening algorithm. Analysis of archived California screening data showed that an arginine cutoff of 50 μM combined with an arginine/ornithine ratio of 1.4 would have resulted in a recall rate of 0.01%. Using an arginine cutoff of 60 μM and an arginine/(phenylalanine x leucine) ratio of 1.4, reportedly used in one screening program, or the R4S Tool Runner, would have resulted in a recall rate of
-
Myocyte-mediated Arginase Expression Controls Hyperargininemia but not Hyperammonemia in Arginase-deficient Mice
Molecular therapy : the journal of the American Society of Gene Therapy, 2014Co-Authors: Jennifer Kasten, Wayne W Grody, Stephen D. Cederbaum, Ragini Bhargava, Hana Park, Denise S. Tai, Quynh G Nguyen, Stephen D. Hauschka, Gerald S. LipshutzAbstract:Human arginase deficiency is characterized by Hyperargininemia and infrequent episodes of hyperammonemia that cause neurological impairment and growth retardation. We previously developed a neonatal mouse adeno-associated viral vector (AAV) rh10-mediated therapeutic approach with arginase expressed by a chicken β-actin promoter that controlled plasma ammonia and arginine, but hepatic arginase declined rapidly. This study tested a codon-optimized arginase cDNA and compared the chicken β-actin promoter to liver- and muscle-specific promoters. ARG1−/− mice treated with AAVrh10 carrying the liver-specific promoter also exhibited long-term survival and declining hepatic arginase accompanied by the loss of AAV episomes during subsequent liver growth. Although arginase expression in striated muscle was not expected to counteract hyperammonemia, due to muscle's lack of other urea cycle enzymes, we hypothesized that the postmitotic phenotype in muscle would allow vector genomes to persist, and hence contribute to decreased plasma arginine. As anticipated, ARG1−/− neonatal mice treated with AAVrh10 carrying a modified creatine kinase-based muscle-specific promoter did not survive longer than controls; however, their plasma arginine levels remained normal when animals were hyperammonemic. These data imply that plasma arginine can be controlled in arginase deficiency by muscle-specific expression, thus suggesting an alternative approach to utilizing the liver for treating Hyperargininemia.
-
lethal phenotype in conditional late onset arginase 1 deficiency in the mouse
Molecular Genetics and Metabolism, 2013Co-Authors: Jennifer Kasten, Wayne W Grody, Peter Paul De Deyn, B Marescau, Ragini Bhargava, Hana Park, Denise Tai, James A Byrne, Lisa Schlichting, Stephen D. CederbaumAbstract:Abstract Human arginase deficiency is characterized by Hyperargininemia and infrequent episodes of hyperammonemia, which lead to neurological impairment with spasticity, loss of ambulation, seizures, and severe mental and growth retardation; uncommonly, patients suffer early death from this disorder. In a murine targeted knockout model, onset of the phenotypic abnormality is heralded by weight loss at around day 15, and death occurs typically by postnatal day 17 with Hyperargininemia and markedly elevated ammonia. This discrepancy between the more attenuated juvenile-onset human disease and the lethal neonatal murine model has remained suboptimal for studying and developing therapy for the more common presentation of arginase deficiency. These investigations aimed to address this issue by creating an adult conditional knockout mouse to determine whether later onset of arginase deficiency also resulted in lethality. Animal survival and ammonia levels, body weight, circulating amino acids, and tissue arginase levels were examined as outcome parameters after widespread Cre- recombinase activation in a conditional knockout model of arginase 1 deficiency. One hundred percent of adult female and 70% of adult male mice died an average of 21.0 and 21.6 days, respectively, after the initiation of tamoxifen administration. Animals demonstrated elevated circulating ammonia and arginine at the onset of phenotypic abnormalities. In addition, brain and liver amino acids demonstrated abnormalities. These studies demonstrate that (a) the absence of arginase in adult animals results in a disease profile (leading to death) similar to that of the targeted knockout and (b) the phenotypic abnormalities seen in the juvenile-onset model are not exclusive to the age of the animal but instead to the biochemistry of the disorder. This adult model will be useful for developing gene- and cell-based therapies for this disorder that will not be limited by the small animal size of neonatal therapy and for developing a better understanding of the characteristics of Hyperargininemia.
-
increased plasma and tissue guanidino compounds in a mouse model of Hyperargininemia
Molecular Genetics and Metabolism, 2008Co-Authors: Joshua L Deignan, Ramaswamy K Iyer, Stephen D. Cederbaum, Peter Paul De Deyn, B Marescau, Justin C Livesay, Wayne W GrodyAbstract:In humans, arginase I (AI)-deficiency results in Hyperargininemia, a metabolic disorder with symptoms of progressive neurological and intellectual impairment, spasticity, persistent growth retardation, and episodic hyperammonemia. A deficiency of arginase II (AII) has never been detected and the clinical disorder, if any, associated with its deficiency has not been defined. Since the spasticity and paucity of hyperammonemic crises seen in human AI-deficient patients are not features of the other urea cycle disorders, the likelihood of ammonia as the main neuropathogenic agent becomes extremely low, and the modest elevations of arginine seen in the brains of our mouse model of Hyperargininemia make it an unlikely candidate as well. Specific guanidino compounds, direct or indirect metabolites of arginine, are elevated in the blood of patients with uremia. Other guanidino compounds are also increased in plasma and cerebrospinal fluid of hyperargininemic patients making them plausible as neurotoxins in these disorders. We analyzed several guanidino compounds in our arginase single and double knockout animals and found that alpha-keto-delta-guanidinovaleric acid, alpha-N-acetylarginine, and argininic acid were increased in the brain tissue from the AI knockout and double knockout animals. Several compounds were also increased in the plasma, liver, and kidneys. This is the first time that several of the guanidino compounds have been shown to be elevated in the brain tissue of an arginase-deficient mammal, and it further supports their possible role as the neuropathogenic agents responsible for the complications seen in arginase deficiency.
Angela T S Wyse - One of the best experts on this subject based on the ideXlab platform.
-
Hyperargininemia and renal oxidative stress prevention by antioxidants and ng nitro l arginine methyl ester
Journal of Biochemical and Molecular Toxicology, 2017Co-Authors: Daniela Delwing De Lima, Debora Delwing Dal Magro, Cindy Lais Pett Vieira, Gislaine Maria Marestoni Grola, Debora Adriana Fischer, Angela T S WyseAbstract:We investigated the in vitro and in vivo effects of arginine (Arg) on thiobarbituric acid-reactive substances (TBA-RS) and on the activities of catalase (CAT), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) in renal tissues of rats. We also studied the influence of antioxidants (α-tocopherol plus ascorbic acid) and nitric oxide synthase inhibitor NG -nitro-l-arginine methyl ester (l-NAME) on the effects elicited by Arg. Results showed that Arg in vitro (1.5 mM) decreased SOD activity and increased the levels of TBA-RS in the renal medulla. Acute administration of Arg [0.8 g/kg, intraperitoneal injection] decreased CAT activity, increased SOD activity and TBA-RS levels in the renal medulla, and decreased CAT activity in the renal cortex of rats. Most results were prevented by antioxidants and/or l-NAME. Data indicate that Arg causes an oxidative imbalance in the renal tissues studied; however, in the presence of antioxidants and l-NAME, some of these alterations in oxidative stress were prevented.
-
effect of n acetylarginine a metabolite accumulated in Hyperargininemia on parameters of oxidative stress in rats protective role of vitamins and l name
Cell Biochemistry and Function, 2014Co-Authors: Simone Sasso, Angela T S Wyse, Debora Delwing Dal Magro, Leticia Dalmedico, Daniela Delwing De LimaAbstract:In the present investigation, we initially evaluated the in vitro effect of N-acetylarginine on thiobarbituric acid-reactive substances (TBA-RS), total sulfhydryl content and on the activities of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the blood, kidney and liver of rats. Results showed that N-acetylarginine, at a concentration of 5.0 μM, decreased the activity of CAT in erythrocytes, enhanced TBA-RS in the renal cortex, decreased CAT and SOD activities in the renal medulla and decreased CAT and increased SOD and GSH-Px activities in the liver of 60-day-old rats. Furthermore, we tested the influence of the antioxidants, trolox and ascorbic acid, as well as of the N(ω) -nitro-L-arginine methyl ester (L-NAME) on the effects elicited by N-acetylarginine on the parameters tested. Antioxidants and L-NAME prevented most of the alterations caused by N-acetylarginine on the oxidative stress parameters evaluated. Data indicate that oxidative stress induction is probably mediated by the generation of NO and/or ONOO(-) and other free radicals because L-NAME and antioxidants prevented the effects caused by N-acetylarginine in the blood, renal tissues and liver of rats. Our findings lend support to a potential therapeutic strategy for this condition, which may include the use of appropriate antioxidants for ameliorating the damage caused by N-acetylarginine.
-
ntpdase and 5 nucleotidase activities of synaptosomes from hippocampus of rats subjected to Hyperargininemia
Neurochemical Research, 2007Co-Authors: Debora Delwing, Daniela Delwing, Manuela De Castilhos Franca Goncalves, Joao Jose Freitas Sarkis, Angela T S WyseAbstract:ATP is an important excitatory neurotransmitter and adenosine acts as a neuromodulatory structure inhibiting neurotransmitters release in the central nervous system. Since the ecto-nucleotidase cascade that hydrolyzes ATP to adenosine is involved in the control of brain functions and previous studies realized in our laboratory have recently reported that acute administration of Arg decreases the NTPDase and 5′-nucleotidase activities of rat blood serum, in the present study we investigated the effect of arginine administration on NTPDase and 5′-nucleotidase activities by synaptosomes from hippocampus of rats. First, sixty-days-old rats were treated with a single or a triple intraperitoneal injection of arginine (0.8 g/Kg) or an equivalent volume of 0.9% saline solution (control) and were killed 1 h later. Second, rats received an intracerebroventricular injection of 1.5 mM arginine solution or saline (5 μL) and were killed 1 h later. We also tested the in vitro effect of arginine (0.1–1.5 mM) on nucleotide hydrolysis in synaptosomes from rat hippocampus. Results showed that intraperitoneal arginine administration did not alter nucleotide hydrolysis. On the other hand, arginine administered intracerebroventricularly reduced ATP (32%), ADP (30%) and AMP (21%) hydrolysis, respectively. In addition, arginine added to the incubation medium, provoked a decrease on ATP (19%), ADP (17%) and AMP (23%) hydrolysis, respectively. Furthermore, kinetic studies showed that the inhibitory effect of arginine was uncompetitive in relation to ATP, ADP and AMP. In conclusion, according to our results it seems reasonable to postulate that arginine alters the cascade involved in the extracellular degradation of ATP to adenosine.
-
in vitro stimulation of oxidative stress in cerebral cortex of rats by the guanidino compounds accumulating in Hyperargininemia
Brain Research, 2001Co-Authors: Angela T S Wyse, Caren Serra Bavaresco, Martine Elisabeth Kienzle Hagen, Daniela Delwing, Clovis Milton Duval Wannmacher, Carlos Severo Dutrafilho, Moacir WajnerAbstract:Hyperargininemia is a metabolic disorder biochemically characterized by tissue accumulation of arginine and other guanidino compounds. Convulsions, lethargy and psychomotor delay or cognitive deterioration are predominant clinical features of this disease. Although neurologic symptoms predominate in this disorder, their pathophysiology is still unknown. In the present study we investigated the in vitro effects of arginine, N-acetylarginine, argininic acid and homoarginine on some oxidative stress parameters in rat brain in the hope to identify a possible mechanism for the brain damage in Hyperargininemia. Chemiluminescence, total radical-trapping antioxidant potential (TRAP), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities were measured in the cerebral cortex of rats in the presence of various concentrations of these compounds. The results showed that all guanidino compounds tested significantly increased chemiluminescence and decreased TRAP at concentrations similar to those observed in the tissue of hyperargininemic patients. Furthermore, these compounds inhibited CAT and GSH-Px activities to varying extents, with GSH-Px activity being more susceptible to their action. In turn, argininic acid inhibited all enzyme activities, and its main action was also directed towards GSH-Px. The results suggest that oxidative stress caused by guanidino compounds may be involved in the brain dysfunction amongst other potential pathophysiological mechanisms observed in Hyperargininemia.
-
in vitro inhibition of na k atpase activity from rat cerebral cortex by guanidino compounds accumulating in Hyperargininemia
Brain Research, 1999Co-Authors: Cleide Goncalves Da Silva, Clovis Milton Duval Wannmacher, Moacir Wajner, Edino Parolo, Emilio L Streck, Angela T S WyseAbstract:Abstract Hyperargininemia is a metabolic disorder biochemically characterized by tissue accumulation of arginine (Arg) and other guanidino compounds (GC). Convulsions, lethargy and psychomotor delay are predominant clinical features of this disease. Considering that some GC are epileptogenic and cause a decrease in membrane fluidity and that Na+,K+-ATPase, a membrane-bound enzyme, is essential for cellular excitability and is decreased in experimental and human epilepsy, in the present study we determined the in vitro effects of Arg, N-acetylarginine (NAA), argininic acid (AA) and homoarginine (HA) on the activity of Na+,K+-ATPase in the synaptic plasma membrane from cerebral cortex of young rats in the hope to identify a possible mechanism for the brain damage in Hyperargininemia. The results showed that all GC tested, except Arg, significantly inhibited Na+,K+-ATPase activity at concentrations similar to those observed in plasma and CSF of patients with Hyperargininemia. In addition, competition between NAA, AA and HA for the binding to the enzyme was observed, suggesting a common binding site for the GC. It is therefore possible that the inhibitory effect of GC on Na+,K+-ATPase may be related to the brain dysfunction observed in Hyperargininemia.
Gerald S. Lipshutz - One of the best experts on this subject based on the ideXlab platform.
-
lipid nanoparticle targeted mrna therapy as a treatment for the inherited metabolic liver disorder arginase deficiency
Proceedings of the National Academy of Sciences of the United States of America, 2019Co-Authors: Brian Truong, Stephen D. Cederbaum, Gabriella Allegri, Kristine E Burke, Johannes Haberle, Paolo Martini, Gerald S. LipshutzAbstract:Arginase deficiency is caused by biallelic mutations in arginase 1 (ARG1), the final step of the urea cycle, and results biochemically in Hyperargininemia and the presence of guanidino compounds, while it is clinically notable for developmental delays, spastic diplegia, psychomotor function loss, and (uncommonly) death. There is currently no completely effective medical treatment available. While preclinical strategies have been demonstrated, disadvantages with viral-based episomal-expressing gene therapy vectors include the risk of insertional mutagenesis and limited efficacy due to hepatocellular division. Recent advances in messenger RNA (mRNA) codon optimization, synthesis, and encapsulation within biodegradable liver-targeted lipid nanoparticles (LNPs) have potentially enabled a new generation of safer, albeit temporary, treatments to restore liver metabolic function in patients with urea cycle disorders, including ARG1 deficiency. In this study, we applied such technologies to successfully treat an ARG1-deficient murine model. Mice were administered LNPs encapsulating human codon-optimized ARG1 mRNA every 3 d. Mice demonstrated 100% survival with no signs of hyperammonemia or weight loss to beyond 11 wk, compared with controls that perished by day 22. Plasma ammonia, arginine, and glutamine demonstrated good control without elevation of guanidinoacetic acid, a guanidino compound. Evidence of urea cycle activity restoration was demonstrated by the ability to fully metabolize an ammonium challenge and by achieving near-normal ureagenesis; liver arginase activity achieved 54% of wild type. Biochemical and microscopic data showed no evidence of hepatotoxicity. These results suggest that delivery of ARG1 mRNA by liver-targeted nanoparticles may be a viable gene-based therapeutic for the treatment of arginase deficiency.
-
45. Rescue of the Functional Alterations of Motor Cortical Circuits in Arginase 1 Deficiency with AAV-Based Gene Therapy
Molecular Therapy, 2016Co-Authors: Gloria Cantero, Xiao-bo Liu, Steven D. Cederbaum, Peyman Golshani, Gerald S. LipshutzAbstract:The urea cycle is the main mechanism for terrestrial mammals to detoxify excess nitrogen. Disorders of the proximal urea cycle characteristically have periodic episodes of hyperammonemia leading to often severe and permanent neurological deterioration & disability. Ammonia has been implicated by compromising potassium buffering of astrocytic membranes and causing clinical neurological abnormalities by impairing cortical inhibition. Complete arginase 1 (Arg1) deficiency, a distal urea cycle disorder, is the least severe of these abnormalities, demonstrating neurological impairment including spasticity, loss of ambulation and seizures; while characterized by the presence of Hyperargininemia, hyperammonemia is not a frequent clinical finding. While mortality is unfortunately common due to acute episodes of hyperammonemia in proximal urea cycle disorders, patients with Hyperargininemia often are long-lived, however, suffering from progressive intellectual disability and spastic diplegia, and the mechanisms underlying the neurological dysfunction are not understood. To gain better insight on how the loss of arginase expression causes dysfunction in the developing brain, and if gene therapy could prevent these abnormalities, we studied how the excitability and functional and anatomical connectivity of motor cortical neurons are altered in the disorder using the murine knockout model. In addition, we examined if AAV expressing Arg1, administered IV on postnatal day 2, could rescue these findings. Results: Single- and double-copy loss of Arg1 caused dose-dependent decreases in intrinsic excitability, dendritic arborization complexity, and synapses in motor cortex layer V neurons. These findings show that 1) the intrinsic excitability of neurons of homozygous Arg1 knockout mice is abnormal and that, unexpectedly, heterozygous neurons (single copy loss) exhibit an intermediate phenotype compared to wild type and homozygous knockouts (double copy loss) (Fig. AFig. A); corresponding loss of Arg1 decreased the frequency of miniature excitatory postsynaptic currents and the amplitude of miniature inhibitory postsynaptic currents; 2) neuronal branching and spine phenotypes differ between genotypes with, unexpectedly, an intermediate phenotype for heterozygotes (Fig. BFig. B); and 3) with electron microscopic analysis and comparison of layer V synapses from arginase 1 knockout, heterozygous, and wild type mice, there is a very low density of excitatory (i.e. asymmetrical) synapses (Fig. CFig. C) in the knockout and decreased number of inhibitory (perisomatic) synapses (i.e. symmetrical) on somata of pyramidal cells, both dramatic findings. Finally, changes in synaptic morphology and abnormal ultrastructural features were found in knockout mice, also suggesting neuronal degeneration and inflammation. Neonatal intravenous administration on the second postnatal day with AAV expressing arginase 1 by a hepatocyte-specific promoter led to a near-resolution of these abnormalities when administered to homozygous Arg1 knockout animals. Summary: Our studies suggest that arginase 1 deficiency leads to severe and specific changes to intrinsic excitability and synaptic connectivity of motor cortical circuits. Importantly, we find that neonatal AAV-based Arg1 gene expression is effective in reversing both the physiological and anatomical hallmarks of the disorder.View Large Image | Download PowerPoint Slide
-
Minimal ureagenesis is necessary for survival in the murine model of Hyperargininemia treated by AAV-based gene therapy
Gene Therapy, 2015Co-Authors: D S Tai, S D Cederbaum, Hana Park, G Cantero, E Chan, M Yudkoff, Gerald S. LipshutzAbstract:Hyperammonemia is less severe in arginase 1 deficiency compared with other urea cycle defects. Affected patients manifest Hyperargininemia and infrequent episodes of hyperammonemia. Patients typically suffer from neurological impairment with cortical and pyramidal tract deterioration, spasticity, loss of ambulation, seizures and intellectual disability; death is less common than with other urea cycle disorders. In a mouse model of arginase I deficiency, the onset of symptoms begins with weight loss and gait instability, which progresses toward development of tail tremor with seizure-like activity; death typically occurs at about 2 weeks of life. Adeno-associated viral vector gene replacement strategies result in long-term survival of mice with this disorder. With neonatal administration of vector, the viral copy number in the liver greatly declines with hepatocyte proliferation in the first 5 weeks of life. Although the animals do survive, it is not known from a functional standpoint how well the urea cycle is functioning in the adult animals that receive adeno-associated virus. In these studies, we administered [1-13C] acetate to both littermate controls and adeno-associated virus-treated arginase 1 knockout animals and examined flux through the urea cycle. Circulating ammonia levels were mildly elevated in treated animals. Arginine and glutamine also had perturbations. Assessment 30 min after acetate administration demonstrated that ureagenesis was present in the treated knockout liver at levels as low at 3.3% of control animals. These studies demonstrate that only minimal levels of hepatic arginase activity are necessary for survival and ureagenesis in arginase-deficient mice and that this level of activity results in control of circulating ammonia. These results may have implications for potential therapy in humans with arginase deficiency.
-
Myocyte-mediated Arginase Expression Controls Hyperargininemia but not Hyperammonemia in Arginase-deficient Mice
Molecular therapy : the journal of the American Society of Gene Therapy, 2014Co-Authors: Jennifer Kasten, Wayne W Grody, Stephen D. Cederbaum, Ragini Bhargava, Hana Park, Denise S. Tai, Quynh G Nguyen, Stephen D. Hauschka, Gerald S. LipshutzAbstract:Human arginase deficiency is characterized by Hyperargininemia and infrequent episodes of hyperammonemia that cause neurological impairment and growth retardation. We previously developed a neonatal mouse adeno-associated viral vector (AAV) rh10-mediated therapeutic approach with arginase expressed by a chicken β-actin promoter that controlled plasma ammonia and arginine, but hepatic arginase declined rapidly. This study tested a codon-optimized arginase cDNA and compared the chicken β-actin promoter to liver- and muscle-specific promoters. ARG1−/− mice treated with AAVrh10 carrying the liver-specific promoter also exhibited long-term survival and declining hepatic arginase accompanied by the loss of AAV episomes during subsequent liver growth. Although arginase expression in striated muscle was not expected to counteract hyperammonemia, due to muscle's lack of other urea cycle enzymes, we hypothesized that the postmitotic phenotype in muscle would allow vector genomes to persist, and hence contribute to decreased plasma arginine. As anticipated, ARG1−/− neonatal mice treated with AAVrh10 carrying a modified creatine kinase-based muscle-specific promoter did not survive longer than controls; however, their plasma arginine levels remained normal when animals were hyperammonemic. These data imply that plasma arginine can be controlled in arginase deficiency by muscle-specific expression, thus suggesting an alternative approach to utilizing the liver for treating Hyperargininemia.
Wayne W Grody - One of the best experts on this subject based on the ideXlab platform.
-
Myocyte-mediated Arginase Expression Controls Hyperargininemia but not Hyperammonemia in Arginase-deficient Mice
Molecular therapy : the journal of the American Society of Gene Therapy, 2014Co-Authors: Jennifer Kasten, Wayne W Grody, Stephen D. Cederbaum, Ragini Bhargava, Hana Park, Denise S. Tai, Quynh G Nguyen, Stephen D. Hauschka, Gerald S. LipshutzAbstract:Human arginase deficiency is characterized by Hyperargininemia and infrequent episodes of hyperammonemia that cause neurological impairment and growth retardation. We previously developed a neonatal mouse adeno-associated viral vector (AAV) rh10-mediated therapeutic approach with arginase expressed by a chicken β-actin promoter that controlled plasma ammonia and arginine, but hepatic arginase declined rapidly. This study tested a codon-optimized arginase cDNA and compared the chicken β-actin promoter to liver- and muscle-specific promoters. ARG1−/− mice treated with AAVrh10 carrying the liver-specific promoter also exhibited long-term survival and declining hepatic arginase accompanied by the loss of AAV episomes during subsequent liver growth. Although arginase expression in striated muscle was not expected to counteract hyperammonemia, due to muscle's lack of other urea cycle enzymes, we hypothesized that the postmitotic phenotype in muscle would allow vector genomes to persist, and hence contribute to decreased plasma arginine. As anticipated, ARG1−/− neonatal mice treated with AAVrh10 carrying a modified creatine kinase-based muscle-specific promoter did not survive longer than controls; however, their plasma arginine levels remained normal when animals were hyperammonemic. These data imply that plasma arginine can be controlled in arginase deficiency by muscle-specific expression, thus suggesting an alternative approach to utilizing the liver for treating Hyperargininemia.
-
lethal phenotype in conditional late onset arginase 1 deficiency in the mouse
Molecular Genetics and Metabolism, 2013Co-Authors: Jennifer Kasten, Wayne W Grody, Peter Paul De Deyn, B Marescau, Ragini Bhargava, Hana Park, Denise Tai, James A Byrne, Lisa Schlichting, Stephen D. CederbaumAbstract:Abstract Human arginase deficiency is characterized by Hyperargininemia and infrequent episodes of hyperammonemia, which lead to neurological impairment with spasticity, loss of ambulation, seizures, and severe mental and growth retardation; uncommonly, patients suffer early death from this disorder. In a murine targeted knockout model, onset of the phenotypic abnormality is heralded by weight loss at around day 15, and death occurs typically by postnatal day 17 with Hyperargininemia and markedly elevated ammonia. This discrepancy between the more attenuated juvenile-onset human disease and the lethal neonatal murine model has remained suboptimal for studying and developing therapy for the more common presentation of arginase deficiency. These investigations aimed to address this issue by creating an adult conditional knockout mouse to determine whether later onset of arginase deficiency also resulted in lethality. Animal survival and ammonia levels, body weight, circulating amino acids, and tissue arginase levels were examined as outcome parameters after widespread Cre- recombinase activation in a conditional knockout model of arginase 1 deficiency. One hundred percent of adult female and 70% of adult male mice died an average of 21.0 and 21.6 days, respectively, after the initiation of tamoxifen administration. Animals demonstrated elevated circulating ammonia and arginine at the onset of phenotypic abnormalities. In addition, brain and liver amino acids demonstrated abnormalities. These studies demonstrate that (a) the absence of arginase in adult animals results in a disease profile (leading to death) similar to that of the targeted knockout and (b) the phenotypic abnormalities seen in the juvenile-onset model are not exclusive to the age of the animal but instead to the biochemistry of the disorder. This adult model will be useful for developing gene- and cell-based therapies for this disorder that will not be limited by the small animal size of neonatal therapy and for developing a better understanding of the characteristics of Hyperargininemia.
-
increased plasma and tissue guanidino compounds in a mouse model of Hyperargininemia
Molecular Genetics and Metabolism, 2008Co-Authors: Joshua L Deignan, Ramaswamy K Iyer, Stephen D. Cederbaum, Peter Paul De Deyn, B Marescau, Justin C Livesay, Wayne W GrodyAbstract:In humans, arginase I (AI)-deficiency results in Hyperargininemia, a metabolic disorder with symptoms of progressive neurological and intellectual impairment, spasticity, persistent growth retardation, and episodic hyperammonemia. A deficiency of arginase II (AII) has never been detected and the clinical disorder, if any, associated with its deficiency has not been defined. Since the spasticity and paucity of hyperammonemic crises seen in human AI-deficient patients are not features of the other urea cycle disorders, the likelihood of ammonia as the main neuropathogenic agent becomes extremely low, and the modest elevations of arginine seen in the brains of our mouse model of Hyperargininemia make it an unlikely candidate as well. Specific guanidino compounds, direct or indirect metabolites of arginine, are elevated in the blood of patients with uremia. Other guanidino compounds are also increased in plasma and cerebrospinal fluid of hyperargininemic patients making them plausible as neurotoxins in these disorders. We analyzed several guanidino compounds in our arginase single and double knockout animals and found that alpha-keto-delta-guanidinovaleric acid, alpha-N-acetylarginine, and argininic acid were increased in the brain tissue from the AI knockout and double knockout animals. Several compounds were also increased in the plasma, liver, and kidneys. This is the first time that several of the guanidino compounds have been shown to be elevated in the brain tissue of an arginase-deficient mammal, and it further supports their possible role as the neuropathogenic agents responsible for the complications seen in arginase deficiency.
-
arginase induction by sodium phenylbutyrate in mouse tissues and human cell lines
Molecular Genetics and Metabolism, 2007Co-Authors: Rita M Kern, Ramaswamy K Iyer, Wayne W Grody, Z Yang, Stephen D. CederbaumAbstract:Abstract Hyperargininemia is a urea cycle disorder caused by mutations in the gene for arginase I (AI) resulting in elevated blood arginine and ammonia levels. Sodium phenylacetate and a precursor, sodium phenylbutyrate (NaPB) have been used to lower ammonia, conjugating glutamine to produce phenylacetylglutamine which is excreted in urine. The elevated arginine levels induce the second arginase (AII) in patient kidney and kidney tissue culture. It has been shown that NaPB increases expression of some target genes and we tested its effect on arginase induction. Eight 9-week old male mice fed on chow containing 7.5 g NaPB/kg rodent chow and drank water with 10 g NaPB/L, and four control mice had a normal diet. After one week all mice were sacrificed. The arginase specific activities for control and NaPB mice, respectively, were 38.2 and 59.4 U/mg in liver, 0.33 and 0.42 U/mg in kidney, and 0.29 and 1.19 U/mg in brain. Immunoprecipitation of arginase in each tissue with AI and AII antibodies showed the activity induced by NaPB is mostly AI. AII may also be induced in kidney. AI accounts for the fourfold increased activity in brain. In some cell lines, NaPB increased arginase activity up to fivefold depending on dose (1–5 mM) and exposure time (2–5 days); control and NaPB activities, respectively, are: erythroleukemia, HEL, 0.06 and 0.31 U/mg, and K562, 0.46 and 1.74 U/mg; embryonic kidney, HEK293, 1.98 and 3.58 U/mg; breast adenocarcinoma, MDA-MB-468, 1.11 and 4.06 U/mg; and prostate adenocarcinoma, PC-3, 0.55 and 3.20 U/mg. In MDA-MB-468 and HEK most, but not all, of the induced activity is AI. These studies suggest that NaPB may induce AI when used to treat urea cycle disorders. It is relatively less useful in AI deficiency, although it could have some effect in those patients with missense mutations.
-
mouse model for human arginase deficiency
Molecular and Cellular Biology, 2002Co-Authors: Ramaswamy K Iyer, Rita M Kern, Wayne W Grody, Paul K Yoo, Nora Rozengurt, Rosemarie W Tsoa, William E Obrien, Stephen D. CederbaumAbstract:Deficiency of liver arginase (AI) causes Hyperargininemia (OMIM 207800), a disorder characterized by progressive mental impairment, growth retardation, and spasticity and punctuated by sometimes fatal episodes of hyperammonemia. We constructed a knockout mouse strain carrying a nonfunctional AI gene by homologous recombination. Arginase AI knockout mice completely lacked liver arginase (AI) activity, exhibited severe symptoms of hyperammonemia, and died between postnatal days 10 and 14. During hyperammonemic crisis, plasma ammonia levels of these mice increased >10-fold compared to those for normal animals. Livers of AI-deficient animals showed hepatocyte abnormalities, including cell swelling and inclusions. Plasma amino acid analysis showed the mean arginine level in knockouts to be approximately fourfold greater than that for the wild type and threefold greater than that for heterozygotes; the mean proline level was approximately one-third and the ornithine level was one-half of the proline and ornithine levels, respectively, for wild-type or heterozygote mice—understandable biochemical consequences of arginase deficiency. Glutamic acid, citrulline, and histidine levels were about 1.5-fold higher than those seen in the phenotypically normal animals. Concentrations of the branched-chain amino acids valine, isoleucine, and leucine were 0.4 to 0.5 times the concentrations seen in phenotypically normal animals. In summary, the AI-deficient mouse duplicates several pathobiological aspects of the human condition and should prove to be a useful model for further study of the disease mechanism(s) and to explore treatment options, such as pharmaceutical administration of sodium phenylbutyrate and/or ornithine and development of gene therapy protocols.
Jeanlouls Dhondt - One of the best experts on this subject based on the ideXlab platform.
-
Hyperargininemia intellectual and motor improvement related to changes in biochemical data
The Journal of Pediatrics, 1991Co-Authors: Bart Marescau, Marie Lambert, Manon Desjardins, Marie Laberge, Jeanlouls DhondtAbstract:Hyperargininemia is an inherited urea cycle disorder resulting from deficiency of arginase (EC 3.5.3.1). The clinical picture includes progressive spastic diplegia, mental retardation, and epilepsy. Acute episodes of vomiting, ataxia, and lethargy or agitation can also occur. Since the disorder was first described in 1969,1 various treatments have been proposed, including restriction of protein intake, oral administration of an amino acid mixture, therapy with sodium benzoate or sodium phenylacetate or both, and enzyme replacement. However, only five patients have been described who have shown sustained improvement in motor and mental abilities during therapy. 2-4 Another patient had improvement in auditory brain-stem evoked potentials during treatment. 5 The mechanisms responsible for neurologic damage are unknown but are unlikely to be due to hyperammonemia alone. Arginine and its guanidino metabolites are candidate neurotoxins. 6,7 A disturbance of central monoamine metabolism has also been described in one patient.8, 9 In this article, we describe the clinical course of a girl with Hyperargininemia whose condition progressively improved during a 24-month period of strict metabolic control. We also present the results of serial determinations of ammonium, arginine, and its guanidino metabolites and of neuro-