ABSTRACT
Patients with urea cycle disorders intermittently develop episodes of decompensation with hyperammonemia. Although such an episode is often associated with starvation and catabolism, its molecular basis is not fully understood. First, we attempted to elucidate the mechanism of such starvation-associated hyperammonemia. Using a mouse embryonic fibroblast (MEF) culture system, we found that glucose starvation increases ammonia production, and that this increase is associated with enhanced glutaminolysis. These results led us to focus on α-ketoglutarate (AKG), a glutamate dehydrogenase inhibitor, and a major anaplerotic metabolite. Hence, we sought to determine the effect of dimethyl α-ketoglutarate (DKG), a cell-permeable AKG analog, on MEFs and found that DKG mitigates ammonia production primarily by reducing flux through glutamate dehydrogenase. We also verified that DKG reduces ammonia in an NH4 Cl-challenged hyperammonemia mouse model and observed that DKG administration reduces plasma ammonia concentration to 22.8% of the mean value for control mice that received only NH4 Cl. In addition, we detected increases in ornithine concentration and in the ratio of ornithine to arginine following DKG treatment. We subsequently administered DKG intravenously to a newborn pig with hyperammonemia due to ornithine transcarbamylase deficiency and found that blood ammonia concentration declined significantly over time. We determined that this effect is associated with facilitated reductive amination and glutamine synthesis. Our present data indicate that energy starvation triggers hyperammonemia through enhanced glutaminolysis and that DKG reduces ammonia accumulation via pleiotropic mechanisms both in vitro and in vivo. Thus, cell-permeable forms of AKG are feasible candidates for a novel hyperammonemia treatment.
Subject(s)
Hyperammonemia , Ornithine Carbamoyltransferase Deficiency Disease , Mice , Animals , Swine , Hyperammonemia/drug therapy , Hyperammonemia/metabolism , Glutamine/metabolism , Ammonia , Glutamate Dehydrogenase , Fibroblasts/metabolism , OrnithineABSTRACT
IMPORTANCE: Sapropterin hydrochloride, a natural coenzyme (6R-tetrahydrobiopterin) of phenylalanine hydroxylase, was first approved as a treatment for tetrahydrobiopterin deficiency in 1992 in Japan, and was then approved as a treatment for a tetrahydrobiopterin-responsive hyperphenylalaninemia in 2007 and 2008, in the USA and Japan, respectively. Guidelines are required on the proper use of sapropterin hydrochloride for tetrahydrobiopterin-responsive hyperphenylalaninemia. OBSERVATIONS: It is recommended that tetrahydrobiopterin-responsive hyperphenylalaninemia should be diagnosed in all cases of hyperphenylalaninemia, including phenylketonuria, by tetrahydrobiopterin administration tests rather than by phenotype or blood phenylalanine levels. CONCLUSIONS AND RELEVANCE: If tetrahydrobiopterin-responsive hyperphenylalaninemia is diagnosed, all ages can be treated with sapropterin hydrochloride. Although there are reports that sapropterin hydrochloride is effective and safe for the prevention of maternal phenylketonuria, further investigation is required.
Subject(s)
Biopterins/analogs & derivatives , Phenylketonurias , Biopterins/therapeutic use , Female , Humans , Japan , Phenotype , Phenylalanine , Phenylalanine Hydroxylase , Phenylketonuria, Maternal/prevention & control , Phenylketonurias/diagnosis , Phenylketonurias/therapy , PregnancyABSTRACT
We report the case of a family with late-onset ornithine transcarbamylase deficiency (OTCD). Several family members had died from OTCD, and the c.221G>A, p.Lys221Lys mutation was detected at the 3' end of exon 6 of OTC in the X-chromosome of some members. We provided genetic counseling on pregnancy, delivery, and neonate management to a 4th-generation female carrier and decided on metabolic management of her child from birth. Two male patients were diagnosed with late-onset OTCD on the basis of blood amino acid and genetic analysis, and they received arginine supplementation from the asymptomatic, early neonatal period. These children grew and developed normally, without decompensation. Patients with late-onset OTCD can and should be diagnosed and treated in the early neonatal period, especially those from families already diagnosed with late-onset OTCD, and family members must be provided with genetic counseling.
Subject(s)
DNA/genetics , Genetic Therapy/methods , Mutation , Ornithine Carbamoyltransferase Deficiency Disease/therapy , Ornithine Carbamoyltransferase/genetics , Age of Onset , Child , Child, Preschool , DNA Mutational Analysis , Female , Humans , Infant , Infant, Newborn , Male , Ornithine Carbamoyltransferase/metabolism , Ornithine Carbamoyltransferase Deficiency Disease/diagnosis , Ornithine Carbamoyltransferase Deficiency Disease/genetics , Pedigree , PregnancyABSTRACT
Citrin-deficient children and adolescents between adult-onset type II citrullinemia and neonatal intrahepatic cholestasis by citrin deficiency do not have clear clinical features except for unusual diet of high-fat, high-protein, and low-carbohydrate food. The aims of the present study are to characterize fatigue and quality of life (QOL) in citrin-deficient patients during adaptation and compensation stage, and to define the relationship between fatigue and QOL. The study subjects were 55 citrin-deficient patients aged 1-22years (29 males) and 54 guardians. Fatigue was evaluated by self-reports and proxy-reports of the PedsQL Multidimensional Fatigue Scale. QOL was evaluated by the PedsQL Generic Core Scales. Both scale scores were significantly lower in child self-reports (p<0.01 and p<0.05, respectively) and parent proxy-reports (p<0.01 and p<0.01, respectively) than those of healthy children. Citrin-deficient patients with scores of 50 percentile or less of healthy children constituted 67.5% of the sample for the Fatigue Scale and 68.4% for the Generic Core Scales. The PedsQL Fatigue Scale correlated with the Generic Core Scales for both the patients (r=0.56) and parents reports (r=0.71). Assessments by the patients and their parents showed moderate agreement. Parents assessed the condition of children more favorably than their children. The study identified severe fatigue and impaired QOL in citrin-deficient patients during the silent period, and that such children perceive worse fatigue and poorer QOL than those estimated by their parents. The results stress the need for active involvement of parents and medical staff in the management of citrin-deficient patients during the silent period.
Subject(s)
Adaptation, Physiological , Carbohydrate Metabolism , Citrullinemia/metabolism , Citrullinemia/pathology , Fatigue/metabolism , Adolescent , Calcium-Binding Proteins/deficiency , Child , Child, Preschool , Citrullinemia/therapy , Diet, High-Fat , Fatigue/pathology , Fatigue/therapy , Female , Humans , Infant , Infant, Newborn , Male , Organic Anion Transporters/deficiency , Quality of Life , Young AdultABSTRACT
Kabuki syndrome is a congenital anomaly syndrome characterized by developmental delay, intellectual disability, specific facial features including long palpebral fissures and ectropion of the lateral third of the lower eyelids, prominent digit pads, and skeletal and visceral abnormalities. Mutations in MLL2 and KDM6A cause Kabuki syndrome. We screened 81 individuals with Kabuki syndrome for mutations in these genes by conventional methods (n = 58) and/or targeted resequencing (n = 45) or whole exome sequencing (n = 5). We identified a mutation in MLL2 or KDM6A in 50 (61.7%) and 5 (6.2%) cases, respectively. Thirty-five MLL2 mutations and two KDM6A mutations were novel. Non-protein truncating-type MLL2 mutations were mainly located around functional domains, while truncating-type mutations were scattered through the entire coding region. The facial features of patients in the MLL2 truncating-type mutation group were typical based on those of the 10 originally reported patients with Kabuki syndrome; those of the other groups were less typical. High arched eyebrows, short fifth finger, and hypotonia in infancy were more frequent in the MLL2 mutation group than in the KDM6A mutation group. Short stature and postnatal growth retardation were observed in all individuals with KDM6A mutations, but in only half of the group with MLL2 mutations.
Subject(s)
Abnormalities, Multiple/genetics , DNA-Binding Proteins/genetics , Face/abnormalities , Hematologic Diseases/genetics , Histone Demethylases/genetics , Mutation , Neoplasm Proteins/genetics , Nuclear Proteins/genetics , Vestibular Diseases/genetics , Abnormalities, Multiple/diagnosis , Adolescent , Adult , Amino Acid Substitution , Child , Child, Preschool , Exome , Facies , Female , Genetic Association Studies , Hematologic Diseases/diagnosis , High-Throughput Nucleotide Sequencing , Humans , Infant , Infant, Newborn , Male , Mutation Rate , Phenotype , Vestibular Diseases/diagnosis , X Chromosome Inactivation , Young AdultABSTRACT
Recently, plasma globotriaosylsphingosine (lyso-Gb3) has attracted attention as a biomarker of Fabry disease. However, we found a subset of Fabry disease patients who did not show any increase in the plasma lyso-Gb3 concentration, although other patients exhibited apparent enhancement of it. This subset predominantly exhibited the clinical phenotype of later-onset Fabry disease, and gene analysis revealed that the patients harbored the M296I mutation common to Japanese Fabry patients. This amino acid substitution is predicted to cause a small conformational change on the surface of the α-galactosidase A molecule, resulting in residual enzyme activity. Plasma lyso-Gb3 is a good biomarker of Fabry disease but care should be taken when it is used for a definitive diagnosis.
Subject(s)
Fabry Disease/diagnosis , Glycolipids/blood , Sphingolipids/blood , alpha-Galactosidase/genetics , Adult , Amino Acid Substitution , Asian People , Biomarkers/blood , Child , Child, Preschool , Fabry Disease/enzymology , Fabry Disease/genetics , Female , Humans , Isoleucine/chemistry , Isoleucine/genetics , Male , Methionine/chemistry , Methionine/genetics , Middle Aged , Mutation , Phenotype , alpha-Galactosidase/chemistry , alpha-Galactosidase/metabolismABSTRACT
Urea cycle disorders (UCDs) are one of the most frequently inherited metabolic diseases in Japan, with an estimated prevalence of 1 per 50,000 live births. Here, we investigated the clinical manifestations, treatment, and prognosis of 177 patients with UCDs who were evaluated and treated from January 1999 to March 2009. These included 77 cases of neonatal-onset UCDs and 91 cases of late-onset UCDs. The most common UCD was ornithine transcarbamylase deficiency (OTCD), which accounted for 116 out of 177 patients. This result is similar to a previous study performed between 1978 and 1995 in Japan: OTCD accounted for about two-thirds of the total number of UCD cases. We studied the relationship between prognosis and the peak blood ammonia level at the onset in 151 UCD patients. Compared with a previous survey conducted in Japan, we found that a greater number of patients survived without any mental retardation despite their peak blood ammonia levels being greater than 360 µmol/l. The 5-year survival rate of patients with OTCD improved to 86% for those with the neonatal-onset type and to 92% for those with the late-onset type. We hypothesize that the increased survival rate is due to early diagnosis and better treatments that are now available in Japan. It is very important to diagnose and treat UCDs, especially OTCD, when the blood ammonia levels in patients are low. The outcome in patients with low blood ammonia levels was better than that in patients with high blood ammonia levels.
Subject(s)
Ammonia/blood , Ammonia/metabolism , Urea Cycle Disorders, Inborn/diagnosis , Urea/metabolism , Age of Onset , Female , Humans , Japan , Male , Ornithine Carbamoyltransferase Deficiency Disease/blood , Ornithine Carbamoyltransferase Deficiency Disease/diagnosis , Ornithine Carbamoyltransferase Deficiency Disease/drug therapy , Ornithine Carbamoyltransferase Deficiency Disease/metabolism , Prognosis , Survival Rate , Treatment Outcome , Urea Cycle Disorders, Inborn/blood , Urea Cycle Disorders, Inborn/drug therapy , Urea Cycle Disorders, Inborn/metabolismABSTRACT
We assessed whether laronidase (recombinant human α-L-iduronidase) replacement therapy could improve left ventricular (LV) myocardial function in a 49-year-old woman with mucopolysaccharidosis I (MPS I) and valvular heart disease. After 6 months of laronidase treatment, the concentration of urinary uron acid decreased by 78.8%. Hepatosplenomegaly improved and LV weight decreased by 19.6%. LV ejection fraction assessed by two-dimensional echocardiogram did not change after laronidase treatment. However, in two-dimensional ultrasound speckle tracking imaging method, LV myocardial longitudinal strain (shortening ratio) increased from -13.2 to -17.4%. LV myocardial radial strain (thickening ratio) increased from 26.6 to 83.4%. LV myocardial torsion increased from +6 to +18°. These indexes of myocardial function were normalized after laronidase treatment. Thus, our findings were a first report that laronidase treatment had a beneficial effect on LV myocardial function in an adult patient with MPS I.
Subject(s)
Enzyme Replacement Therapy , Heart Ventricles/drug effects , Iduronidase/pharmacology , Mucopolysaccharidosis I/therapy , Female , Heart Ventricles/physiopathology , Humans , Iduronidase/therapeutic use , Liver/drug effects , Middle Aged , Mucopolysaccharidosis I/pathology , Organ Size/drug effects , Spleen/drug effects , Time Factors , Treatment OutcomeABSTRACT
Few studies have looked at optimal or acceptable serum phenylalanine levels in later life in patients with phenylketonuria (PKU). This study examined the oxidative stress status of adolescents and adults with PKU. Forty PKU patients aged over fifteen years were enrolled, and were compared with thirty age-matched controls. Oxidative stress markers, anti-oxidant enzyme activities in erythrocytes, and blood anti-oxidant levels were examined. Nitric oxide (NO) production was also examined as a measure of oxidative stress. Plasma thiobarbituric acid reactive species and serum malondialdehyde-modified LDL levels were significantly higher in PKU patients than control subjects, and correlated significantly with serum phenylalanine level (P<0.01). Plasma total anti-oxidant reactivity levels were significantly lower in the patient group, and correlated negatively with phenylalanine level (P<0.001). Erythrocyte superoxide dismutase and catalase activities were higher and correlated significantly with phenylalanine level (P<0.01). Glutathione peroxidase activity was lower and correlated negatively with phenylalanine level (P<0.001). The oxidative stress score calculated from these six parameters was significantly higher in patients with serum phenylalanine of 700-800 µmol/l. Plasma anti-oxidant substances, beta-carotene, and coenzyme Q(10) were also lower (P<0.001), although the decreases did not correlate significantly with the phenylalanine level. Serum nitrite/nitrate levels, as stable NO products, were higher together with low serum asymmetric dimethylarginine, as an endogenous NO inhibitor. Oxidative stress status is closely linked with serum phenylalanine levels. Phenylalanine level in should be maintained PKU below 700-800 µmol/l even in adult patients.
Subject(s)
Oxidative Stress , Phenylalanine/blood , Phenylketonurias/physiopathology , Adolescent , Adult , Biomarkers/blood , Biomarkers/urine , Erythrocytes/metabolism , Female , Humans , Male , Middle Aged , Nitric Oxide/blood , Phenylalanine/metabolism , Young AdultABSTRACT
BACKGROUND: Recent studies suggest that refractory hypotension from causes other than septicaemia or cardiac failure is common in extremely preterm infants even out of the transitional period. Marked response to low-dose cortisol suggests underlying adrenal insufficiency, although the exact mechanism remains unknown. METHODS: To investigate potential triggers for and related short-term outcomes of early-onset (Subject(s)
Hypotension/drug therapy
, Infant, Premature, Diseases/drug therapy
, Thyroxine/blood
, Thyroxine/therapeutic use
, Age of Onset
, Female
, Gestational Age
, Humans
, Hypotension/diagnosis
, Hypotension/epidemiology
, Incidence
, Infant
, Infant, Newborn
, Infant, Premature
, Infant, Premature, Diseases/diagnosis
, Infant, Premature, Diseases/epidemiology
, Japan/epidemiology
, Male
, Retrospective Studies
, Treatment Outcome
ABSTRACT
The mechanism underlying the development of osteopenia or osteoporosis in longstanding phenylketonuria (PKU) remains to be clarified. We investigated the details of bone metabolism in 21 female and 13 male classical PKU patients aged 20-35 years. Vitamin D (VD), parathyroid hormone (PTH), bone turnover markers, and daily nutrient intake were examined. The patients had lower daily energy and protein intake than did the age-matched controls (22 women, 14 men), but their respective fat, VD, and calcium intake did not differ. Serum 1,25-dihydroxy VD and 25-hydroxy VD levels in female and male patient groups were significantly higher and lower than those in respective control groups (females, P < 0.001; males, P < 0.05 and P < 0.01, respectively). Serum intact PTH levels were significantly higher in the female patient group (P < 0.05). Urinary calcium levels in the patient groups were significantly higher than those of the control subjects (females, P < 0.001; males, P < 0.05). Bone resorption markers were significantly higher in patients than in controls, although bone formation markers were not different. Patient serum levels of osteoprotegerin-inhibiting bone resorption were significantly lower (females, P < 0.001; males, P < 0.01). None of the bone parameters correlated significantly with serum phenylalanine or nutrient intake. PKU patients exhibited lower VD status and more rapid bone resorption despite normal calcium-VD intakes.
Subject(s)
Bone and Bones/metabolism , Phenylketonurias/diagnosis , Phenylketonurias/metabolism , Adult , Bone Diseases, Metabolic/metabolism , Bone Resorption/blood , Bone Resorption/diagnosis , Cross-Sectional Studies , Female , Humans , Infant, Newborn , Male , Neonatal Screening , Parathyroid Hormone/blood , Phenylketonurias/blood , Vitamin D/blood , Young AdultABSTRACT
In 4 young pediatric patients with presymptomatic Wilson disease, we found zinc monotherapy beginning at time of diagnosis to be safe and highly effective for follow-up intervals between 1 and 2 years. Such maintenance therapy with zinc can maintain urinary copper excretion between 1 and 3 µg · kg(-1) · day.
Subject(s)
Hepatolenticular Degeneration/diagnosis , Hepatolenticular Degeneration/drug therapy , Zinc/therapeutic use , Alanine Transaminase/blood , Aspartate Aminotransferases/blood , Asymptomatic Diseases/therapy , Child , Child, Preschool , Copper/blood , Copper/urine , Female , Follow-Up Studies , HumansABSTRACT
We report a case of a 13-year-old boy with arginase 1 deficiency carrying a new variant in ARG1. Sanger sequencing identified the compound heterozygous variants: NM_000045.4: c.365G>A (p.Trp122*)/c.820G>A (p.Asp274Asn). Although not previously reported, the p.Asp274Asn variant is predicted to have strong pathogenicity because it is located in a highly conserved domain in the protein core and arginase activity in the patient was below measurement sensitivity.
ABSTRACT
We performed haplotype analysis using nine single nucleotide polymorphisms in the ornithine transcarbamylase gene to explore the ancestral origins of three mutations associated with late-onset phenotype in male patients: p.R40H, p.R277W and p.Y55D. Overall, 8 haplotypes were defined among 14 families carrying p.R40H, 5 families carrying p.R277W and 2 families with p.Y55D mutations. Of nine Japanese families carrying p.R40H, eight exhibited haplotype (HT)1, whereas the other family harbored HT2. Among three Caucasian families, one Spanish and one Australian family bore HT3; one Austrian family had HT4. Two US patients harbored HT2 and HT4. Among families carrying p.R277W, HT5 was found in one Japanese, one Korean and one US family. Two other US families had HT2 and HT6. Two families carrying p.Y55D, both Japanese, shared HT1. These results indicate that the p.R40H mutation has arisen recurrently in all populations studied, although there is evidence for a founder effect in Japan, with most cases probably sharing a common origin, and to a lesser extent in subjects of European ancestry (HT3). It is evident that p.R277W mutation has recurred in discrete populations. The p.Y55D mutation appears to have arisen from a common ancestor, because this transversion (c.163T>G) occurs rarely.
Subject(s)
Alleles , Mutation , Ornithine Carbamoyltransferase Deficiency Disease/genetics , Ornithine Carbamoyltransferase/genetics , Age of Onset , Asian People , Evolution, Molecular , Gene Frequency , Haplotypes , Humans , Japan , Male , Ornithine Carbamoyltransferase/chemistry , Polymorphism, Single Nucleotide , White PeopleABSTRACT
Osteoporosis-pseudoglioma syndrome (OPS; OMIM 259770) is an autosomal-recessive genetic disorder characterized by severe osteoporosis and visual disturbance from childhood. Biallelic mutations in the low-density lipoprotein receptor-related protein 5 gene (LRP5) have been frequently detected, while a subset of patients had only one or no detectable mutation. We report on the clinical and molecular findings of four unrelated Japanese patients with the syndrome. The four patients had typical skeletal and ocular phenotypes of OPS, namely severe juvenile osteoporosis and early-onset visual disturbance, with or without mental retardation. We undertook standard PCR-based sequencing for LRP5 and found four missense mutations (p.L145F, p.T244M, p.P382L, and p.T552M), one nonsense mutation (p.R1534X), and one splice site mutation (c.1584+1G>A) among four OPS patients. Although three patients had two heterozygous mutations, one had only one heterozygous splice site mutation. In this patient, RT-PCR from lymphocytic RNA demonstrated splice error resulting in 63-bp insertion between exons 7 and 8. Furthermore, the patient was found to have only mutated RT-PCR fragment, implying that a seemingly normal allele did not express LRP5 mRNA. We then conducted custom- designed oligonucleotide tiling microarray analyses targeted to a 600-kb genome region harboring LRP5 and discovered a 7.2-kb microdeletion encompassing exons 22 and 23 of LRP5. We found various types of LRP5 mutations, including an exon-level deletion that is undetectable by standard PCR-based mutation screening. Oligonucleotide tiling microarray seems to be a powerful tool in identifying cryptic structural mutations.
Subject(s)
Glioma/genetics , LDL-Receptor Related Proteins/genetics , Mutation , Osteoporosis/genetics , Adolescent , Adult , Base Sequence , Child , DNA Primers , Female , Humans , Low Density Lipoprotein Receptor-Related Protein-5 , Male , Oligonucleotide Array Sequence Analysis , Pedigree , Reverse Transcriptase Polymerase Chain Reaction , Sequence Deletion , SyndromeABSTRACT
BACKGROUND: Residual dried blood spots (DBS) remaining after routine newborn screening (NBS) tests are candidate specimens for extended uses such as quality assurance and the development of new technology. A trial of NBS using tandem mass-spectrometry was launched in 2004 in Japan. The aim of the present study was to analyze the attitudes of the public, patient families, and medical professionals toward the extended use and long-term storage of residual DBS, and to construct a standardized informational brochure. METHODS: A questionnaire was sent to randomly selected members of the public, members of the Japanese Phenylketonuria (PKU) Association, medical staff of a general hospital, staff of a children's hospital, obstetricians and gynecologists, pediatricians and NBS personnel. Associated responses, which were given in a free comment format, were analyzed by text mining. RESULTS: The awareness ratio of NBS was low in the public (26.6%), but despite this, when a brief explanatory note on NBS was provided, 71.7% of them recognized the necessity of NBS. They were less positive than medical professionals and PKU patient families regarding the extended use of DBS for forensic investigation, for the study of health problems, or long-term storage of residual DBS, regardless of whether these factors affected them personally or not. Among the medical professionals, obstetricians and pediatricians exhibited a higher ratio of negative responses toward the extended use and long-term storage of DBS than others. CONCLUSION: The general public is more conservative than PKU patients and their families or medical professionals about the extended use or long-term storage of residual DBS. Presentation to the public, particularly to couples of childbearing age, of appropriate explanatory information on NBS itself, or the extended use or long-term storage of residual DBS, is recommended.
Subject(s)
Blood Preservation , Blood Specimen Collection , Neonatal Screening/methods , Surveys and Questionnaires , Attitude of Health Personnel , Female , Humans , Infant, Newborn , Japan , Male , Public Opinion , Quality Control , Specimen Handling , Time FactorsABSTRACT
Issues pertinent to patients with phenylketonuria(PKU) in adulthood are presented. Nutritional management policy that is optimal to prevent such nutritional complications as osteoporosis and possible vitamin B12 deficiency in each age group should be considered. Adolescent girls with PKU and their guardians should be informed of the issue of maternal PKU to prevent the condition. Socioeconomical issues also remain to be solved. Most adult patients have felt that medical expense to continue dietary therapy is a significant economical burden, which often leads to withdrawal from the therapy. Buying life insurance may be refused by insurance companies simply because the patients have PKU. Current knowledge on health status of well-controlled PKU patients should be provided to insurance companies.
Subject(s)
Continuity of Patient Care , Phenylketonurias , Adolescent , Adult , Amino Acids, Neutral/administration & dosage , Biopterins/administration & dosage , Biopterins/analogs & derivatives , Diet Therapy , Female , Humans , Infant, Newborn , Insurance, Life , Neonatal Screening , Phenylalanine , Phenylketonuria, Maternal , Phenylketonurias/diagnosis , Phenylketonurias/genetics , Phenylketonurias/therapy , Pregnancy , Socioeconomic Factors , Young AdultABSTRACT
Mucolipidosis (ML) II alpha/beta and III alpha/beta are autosomal recessive diseases caused by a deficiency of alpha and/or beta subunits of the enzyme N-acetylglucosamine-1-phosphotransferase, which is encoded by the GNPTAB gene. We analyzed the GNPTAB gene in 25 ML II and 15 ML III Japanese patients. In most ML II patients, the clinical conditions 'stand alone', 'walk without support' and 'speak single words' were impaired; however, the frequency of 'heart murmur', 'inguinal hernia' and 'hepatomegaly and/or splenomegaly' did not differ between ML II and III patients. We detected mutations in GNPTAB in 73 of 80 alleles. Fourteen new mutations were c.914_915insA, c.2089_2090insC, c.2427delC, c.2544delA, c.2693delA, c.3310delG, c.3388_3389insC+c.3392C>T, c.3428_3429insA, c.3741_3744delAGAA, p.R334L, p.F374L, p.H956Y, p.N1153S and duplication of exon 2. Previously reported mutations were p.Q104X, p.W894X, p.R1189X and c.2715+1G>A causing skipping of exon 13. Homozygotes or compound heterozygotes of nonsense and frameshift mutations contributed to the severe phenotype. p.F374L, p.N1153S and splicing mutations contributed to the attenuated phenotype, although coupled with nonsense mutation. These results show the effective molecular diagnosis of ML II and III and also provide phenotypic prediction. This is the first and comprehensive report of molecular analysis for ML patients of Japanese origin.
Subject(s)
Asian People/genetics , Mucolipidoses/enzymology , Mucolipidoses/genetics , Transferases (Other Substituted Phosphate Groups)/genetics , Base Sequence , DNA Mutational Analysis , Exons/genetics , Gene Duplication , Genotype , Humans , Molecular Sequence Data , Mutation/genetics , Phenotype , Polymorphism, Single Nucleotide/geneticsABSTRACT
Two distinct human light subunits of the heteromeric amino acid transporter, y+LAT-1 coded by SLC7A7 and y+LAT-2 coded by SLC7A6, are both known to induce transport system y+L activity. SLC7A7 has already been identified as the gene responsible for lysinuric protein intolerance (LPI). We successfully identified five novel SLC7A7 variants (S238F, S489P, 1630delC, 1673delG, and IVS3-IVS5del9.7kb) in Japanese patients with LPI by PCR amplification and direct DNA sequencing. In addition, we performed a semi-quantitative expression analysis of SLC7A7 and SLC7A6 in human tissue. In normal tissue, the gene-expression ratio of SLC7A6 to SLC7A7 was high in the brain, muscle, and cultured skin fibroblasts; low in the kidneys and small intestine; and at an intermediate level in peripheral blood leukocytes, the lungs, and cultured lymphoblasts. The gene-expression ratio of SLC7A6 to SLC7A7 in cultured lymphoblasts was significantly different between normal subjects and LPI patients with R410X and/or S238F, where the relative amount of SLC7A7 mRNA was significantly lower and the relative amount of SLC7A6 mRNA was statistically higher in affected lymphoblasts than in normal cells. Expression of SLC7A7 and SLC7A6 may thus be interrelated in cultured lymphoblasts.
Subject(s)
Amino Acid Transport Disorders, Inborn/genetics , Amino Acid Transport System y+L/biosynthesis , Fusion Regulatory Protein 1, Light Chains/biosynthesis , Fusion Regulatory Protein 1, Light Chains/genetics , Mutation , Adolescent , Amino Acid Transport Disorders, Inborn/metabolism , Amino Acid Transport System y+L/genetics , Amino Acid Transport Systems, Basic/biosynthesis , Amino Acid Transport Systems, Basic/genetics , Base Sequence , Cationic Amino Acid Transporter 1/biosynthesis , Cationic Amino Acid Transporter 1/genetics , Cells, Cultured , Child , DNA Mutational Analysis , Female , Genetic Variation , Humans , Japan , Lymphocyte Activation , Lymphocytes/metabolism , Male , Molecular Sequence Data , RNA, Messenger/biosynthesis , Transcription, GeneticABSTRACT
Classical citrullinemia (CTLN1), a rare autosomal recessive disorder, is caused by mutations of the argininosuccinate synthetase (ASS) gene, localized on chromosome 9q34.1. ASS functions as a rate-limiting enzyme in the urea cycle. Previously, we identified 32 mutations in the ASS gene of CTLN1 patients mainly in Japan and the United States, and to date 34 different mutations have been described in 50 families worldwide. In the present study, we report ASS mutations detected in 35 additional CTLN1 families from 11 countries. By analyzing the entire coding sequence and the intron-exon boundaries of the ASS gene using RT-PCR and/or genomic DNA-PCR, we have identified 16 novel mutations (two different 1-bp deletions, a 67-bp insertion, and 13 missense) and have detected 12 known mutations. Altogether, 50 different mutations (seven deletion, three splice site, one duplication, two nonsense, and 37 missense) in 85 CTLN1 families were identified. On the basis of primary sequence comparisons with the crystal structure of E. coli ASS protein, it may be concluded that any of the 37 missense mutations found at 30 different positions led to structural and functional impairments of the human ASS protein. It has been found that three mutations are particularly frequent: IVS6-2A>G in 23 families (Japan: 20 and Korea: three), G390R in 18 families (Turkey: six, U.S.: five, Spain: three, Israel: one, Austria: one, Canada: one, and Bolivia: one), and R304W in 10 families (Japan: nine and Turkey: one). Most mutations of the ASS gene are "private" and are distributed throughout the gene, except for exons 5 and 12-14. It seems that the clinical course of the patients with truncated mutations or the G390R mutation is early-onset/severe. The phenotype of the patients with certain missense mutations (G362V or W179R) is more late-onset/mild. Eight patients with R86H, A118T, R265H, or K310R mutations were adult/late-onset and four of them showed severe symptoms during pregnancy or postpartum. However, it is still difficult to prove the genotype-phenotype correlation, because many patients were compound heterozygotes (with two different mutations), lived in different environments at the time of diagnosis, and/or had several treatment regimes or various knowledge of the disease.