RESUMO
Serine, glycine and other nonessential amino acids are critical for tumour progression, and strategies to limit their availability are emerging as potential therapies for cancer1-3. However, the molecular mechanisms driving this response remain unclear and the effects on lipid metabolism are relatively unexplored. Serine palmitoyltransferase (SPT) catalyses the de novo biosynthesis of sphingolipids but also produces noncanonical 1-deoxysphingolipids when using alanine as a substrate4,5. Deoxysphingolipids accumulate in the context of mutations in SPTLC1 or SPTLC26,7-or in conditions of low serine availability8,9-to drive neuropathy, and deoxysphinganine has previously been investigated as an anti-cancer agent10. Here we exploit amino acid metabolism and the promiscuity of SPT to modulate the endogenous synthesis of toxic deoxysphingolipids and slow tumour progression. Anchorage-independent growth reprogrammes a metabolic network involving serine, alanine and pyruvate that drives the endogenous synthesis and accumulation of deoxysphingolipids. Targeting the mitochondrial pyruvate carrier promotes alanine oxidation to mitigate deoxysphingolipid synthesis and improve spheroid growth, similar to phenotypes observed with the direct inhibition of SPT or ceramide synthesis. Restriction of dietary serine and glycine potently induces the accumulation of deoxysphingolipids while decreasing tumour growth in xenograft models in mice. Pharmacological inhibition of SPT rescues xenograft growth in mice fed diets restricted in serine and glycine, and the reduction of circulating serine by inhibition of phosphoglycerate dehydrogenase (PHGDH) leads to the accumulation of deoxysphingolipids and mitigates tumour growth. The promiscuity of SPT therefore links serine and mitochondrial alanine metabolism to membrane lipid diversity, which further sensitizes tumours to metabolic stress.
Assuntos
Neoplasias/metabolismo , Neoplasias/patologia , Serina/deficiência , Esfingolipídeos/química , Esfingolipídeos/metabolismo , Alanina/biossíntese , Alanina/metabolismo , Alanina/farmacologia , Animais , Adesão Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Dieta , Feminino , Glicina/biossíntese , Glicina/deficiência , Glicina/metabolismo , Glicina/farmacologia , Células HCT116 , Humanos , Lipídeos de Membrana/química , Lipídeos de Membrana/metabolismo , Camundongos , Mitocôndrias/metabolismo , Neoplasias/tratamento farmacológico , Fosfoglicerato Desidrogenase/antagonistas & inibidores , Fosfoglicerato Desidrogenase/metabolismo , Ácido Pirúvico/metabolismo , Serina/sangue , Serina/farmacologia , Serina C-Palmitoiltransferase/antagonistas & inibidores , Serina C-Palmitoiltransferase/metabolismo , Esferoides Celulares/patologia , Esfingolipídeos/biossíntese , Estresse Fisiológico/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Biomarkers that accurately reflect renal function are essential in management of chronic kidney diseases (CKD). However, in children, age/physique and medication often alter established renal biomarkers. We studied whether amino acid enantiomers in body fluids correlate with renal function and whether they are influenced by physique or steroid medication during development. We conducted a prospective study of children 2 to 18 years old with and without CKD. We analyzed associations of serine/asparagine enantiomers in body fluids with major biochemical parameters as well as physique. To study consequences of kidney dysfunction and steroids on serine/asparagine enantiomers, we generated juvenile mice with uninephrectomy, ischemic reperfusion injury, or dexamethasone treatment. We obtained samples from 27 children, of which 12 had CKD due to congenital (n = 7) and perinatal (n = 5) causes. Plasma D-asparagine and the D/L-serine ratio had robust, positive linear associations with serum creatinine and cystatin C, and detected CKD with high sensitivity and specificity, uninfluenced by body size or biochemical parameters. In the animal study, kidney dysfunction increased plasma D-asparagine and the D/L-serine ratio, but dexamethasone treatment did not. Thus, plasma D-asparagine and the D/L-serine ratio can be useful markers for renal function in children.
Assuntos
Asparagina , Biomarcadores , Insuficiência Renal Crônica , Serina , Criança , Animais , Humanos , Asparagina/sangue , Asparagina/metabolismo , Insuficiência Renal Crônica/sangue , Pré-Escolar , Serina/sangue , Camundongos , Masculino , Feminino , Adolescente , Biomarcadores/sangue , Estudos Prospectivos , Dexametasona , Estereoisomerismo , Creatinina/sangue , Rim/metabolismoRESUMO
BACKGROUND: Identifying mechanisms of diseases with complex inheritance patterns, such as macular telangiectasia type 2, is challenging. A link between macular telangiectasia type 2 and altered serine metabolism has been established previously. METHODS: Through exome sequence analysis of a patient with macular telangiectasia type 2 and his family members, we identified a variant in SPTLC1 encoding a subunit of serine palmitoyltransferase (SPT). Because mutations affecting SPT are known to cause hereditary sensory and autonomic neuropathy type 1 (HSAN1), we examined 10 additional persons with HSAN1 for ophthalmologic disease. We assayed serum amino acid and sphingoid base levels, including levels of deoxysphingolipids, in patients who had macular telangiectasia type 2 but did not have HSAN1 or pathogenic variants affecting SPT. We characterized mice with low serine levels and tested the effects of deoxysphingolipids on human retinal organoids. RESULTS: Two variants known to cause HSAN1 were identified as causal for macular telangiectasia type 2: of 11 patients with HSAN1, 9 also had macular telangiectasia type 2. Circulating deoxysphingolipid levels were 84.2% higher among 125 patients with macular telangiectasia type 2 who did not have pathogenic variants affecting SPT than among 94 unaffected controls. Deoxysphingolipid levels were negatively correlated with serine levels, which were 20.6% lower than among controls. Reduction of serine levels in mice led to increases in levels of retinal deoxysphingolipids and compromised visual function. Deoxysphingolipids caused photoreceptor-cell death in retinal organoids, but not in the presence of regulators of lipid metabolism. CONCLUSIONS: Elevated levels of atypical deoxysphingolipids, caused by variant SPTLC1 or SPTLC2 or by low serine levels, were risk factors for macular telangiectasia type 2, as well as for peripheral neuropathy. (Funded by the Lowy Medical Research Institute and others.).
Assuntos
Neuropatias Hereditárias Sensoriais e Autônomas/genética , Mutação , Telangiectasia Retiniana/genética , Serina C-Palmitoiltransferase/genética , Serina/metabolismo , Esfingolipídeos/metabolismo , Adulto , Idoso , Animais , Análise Mutacional de DNA , Modelos Animais de Doenças , Exoma/genética , Feminino , Neuropatias Hereditárias Sensoriais e Autônomas/complicações , Neuropatias Hereditárias Sensoriais e Autônomas/metabolismo , Humanos , Metabolismo dos Lipídeos , Macula Lutea/patologia , Masculino , Camundongos , Pessoa de Meia-Idade , Linhagem , Telangiectasia Retiniana/complicações , Telangiectasia Retiniana/metabolismo , Fatores de Risco , Serina/sangue , Esfingosina/análogos & derivados , Esfingosina/análise , Adulto JovemRESUMO
The prevalence of non-alcoholic fatty liver disease (NAFLD) continues to increase dramatically, and there is no approved medication for its treatment. Recently, we predicted the underlying molecular mechanisms involved in the progression of NAFLD using network analysis and identified metabolic cofactors that might be beneficial as supplements to decrease human liver fat. Here, we first assessed the tolerability of the combined metabolic cofactors including l-serine, N-acetyl-l-cysteine (NAC), nicotinamide riboside (NR), and l-carnitine by performing a 7-day rat toxicology study. Second, we performed a human calibration study by supplementing combined metabolic cofactors and a control study to study the kinetics of these metabolites in the plasma of healthy subjects with and without supplementation. We measured clinical parameters and observed no immediate side effects. Next, we generated plasma metabolomics and inflammatory protein markers data to reveal the acute changes associated with the supplementation of the metabolic cofactors. We also integrated metabolomics data using personalized genome-scale metabolic modeling and observed that such supplementation significantly affects the global human lipid, amino acid, and antioxidant metabolism. Finally, we predicted blood concentrations of these compounds during daily long-term supplementation by generating an ordinary differential equation model and liver concentrations of serine by generating a pharmacokinetic model and finally adjusted the doses of individual metabolic cofactors for future human clinical trials.
Assuntos
Acetilcisteína/administração & dosagem , Carnitina/administração & dosagem , Metabolômica/métodos , Niacinamida/análogos & derivados , Serina/administração & dosagem , Acetilcisteína/sangue , Adulto , Animais , Carnitina/sangue , Suplementos Nutricionais , Quimioterapia Combinada , Voluntários Saudáveis , Humanos , Masculino , Modelos Animais , Niacinamida/administração & dosagem , Niacinamida/sangue , Hepatopatia Gordurosa não Alcoólica/dietoterapia , Medicina de Precisão , Compostos de Piridínio , Ratos , Serina/sangueRESUMO
BACKGROUND: Endometrial cancer is strongly associated with obesity and dysregulation of metabolic factors such as estrogen and insulin signaling are causal risk factors for this malignancy. To identify additional novel metabolic pathways associated with endometrial cancer we performed metabolomic analyses on pre-diagnostic plasma samples from 853 case-control pairs from the European Prospective Investigation into Cancer and Nutrition (EPIC). METHODS: A total of 129 metabolites (acylcarnitines, amino acids, biogenic amines, glycerophospholipids, hexoses, and sphingolipids) were measured by liquid chromatography-mass spectrometry. Conditional logistic regression estimated the associations of metabolites with endometrial cancer risk. An analysis focusing on clusters of metabolites using the bootstrap lasso method was also employed. RESULTS: After adjustment for body mass index, sphingomyelin [SM] C18:0 was positively (OR1SD: 1.18, 95% CI: 1.05-1.33), and glycine, serine, and free carnitine (C0) were inversely (OR1SD: 0.89, 95% CI: 0.80-0.99; OR1SD: 0.89, 95% CI: 0.79-1.00 and OR1SD: 0.91, 95% CI: 0.81-1.00, respectively) associated with endometrial cancer risk. Serine, C0 and two sphingomyelins were selected by the lasso method in >90% of the bootstrap samples. The ratio of esterified to free carnitine (OR1SD: 1.14, 95% CI: 1.02-1.28) and that of short chain to free acylcarnitines (OR1SD: 1.12, 95% CI: 1.00-1.25) were positively associated with endometrial cancer risk. Further adjustment for C-peptide or other endometrial cancer risk factors only minimally altered the results. CONCLUSION: These findings suggest that variation in levels of glycine, serine, SM C18:0 and free carnitine may represent specific pathways linked to endometrial cancer development. If causal, these pathways may offer novel targets for endometrial cancer prevention.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias do Endométrio/diagnóstico , Idoso , Biomarcadores Tumorais/metabolismo , Índice de Massa Corporal , Carnitina/sangue , Carnitina/metabolismo , Estudos de Casos e Controles , Neoplasias do Endométrio/sangue , Neoplasias do Endométrio/epidemiologia , Neoplasias do Endométrio/metabolismo , Feminino , Glicina/sangue , Glicina/metabolismo , Humanos , Metabolômica , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Serina/sangue , Serina/metabolismo , Esfingomielinas/sangue , Esfingomielinas/metabolismoRESUMO
INTRODUCTION: D-Serine, present only in trace amounts in humans, is now recognized as a biomarker of chronic kidney disease (CKD). CKD is heterogeneous in its original kidney diseases, whose diagnoses require kidney biopsy. In this study, we examined whether the intra-body dynamics of D-serine, indexed by its blood and urinary levels, reflects the origin of kidney diseases. METHODS: Patients with six kinds of kidney disease undergoing kidney biopsy were enrolled in a single center. Levels of D- and L-serine were measured using two-dimensional high-performance liquid chromatography. The associations between the origin of kidney diseases and the intra-body dynamics of D-serine were examined using multivariate cluster analyses. RESULTS: Unlike the non-CKD profile, patients with CKD showed broadly-distributed profiles of intra-body dynamics of D-serine. The plasma level of D-serine plays a key role in the detection of kidney diseases, whereas a combination of plasma and urinary levels of D-serine distinguished the origin of CKD, especially lupus nephritis. CONCLUSION: Intra-body dynamics of D-serine have the potential to predict the origin of kidney diseases. Monitoring of D-serine may guide specific treatments for the origin of kidney diseases.
Assuntos
Nefropatias/etiologia , Serina/sangue , Serina/urina , Adulto , Idoso , Feminino , Humanos , Nefropatias/sangue , Nefropatias/urina , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/etiologia , Insuficiência Renal Crônica/urinaRESUMO
D-Amino acids are the recently detected enantiomers of L-amino acids. Accumulating evidence points their potential in solving the long-standing critical problems associated with the management of both chronic and acute kidney diseases. This includes estimating kidney function, early diagnosis and prognosis of chronic kidney disease, and disease monitoring. Among the D-amino acids, D-serine levels in the blood are strongly correlated with the glomerular filtration rate and are useful for estimating the function of the kidney. Urinary D-serine also reflects other conditions. The kidney proximal tubule reabsorbs serine with chiral-selectivity, with D-serine being reabsorbed much less efficiently than L-serine, and urinary excretion of D-serine is sensitive to the presence of kidney diseases. Therefore, assessing the intra-body dynamics of D-serine by measuring its level in blood and urinary excretion can be used to detect kidney diseases and assess pathophysiology. This new concept, the intra-body dynamics of D-serine, can be useful in the comprehensive management of kidney disease.
Assuntos
Insuficiência Renal Crônica/diagnóstico , Insuficiência Renal Crônica/fisiopatologia , Serina/sangue , Serina/urina , Biomarcadores/sangue , Biomarcadores/urina , Taxa de Filtração Glomerular , Humanos , PrognósticoRESUMO
Sericin is a protein component of the silkworm cocoon, and contains a high proportion of L-serine, but it has been mostly disposed of as an industrial waste. However, recent studies have revealed its unique biological functionalities beneficial to human health. This study aimed to evaluate the effect of acute oral intake of sericin on amino acid and neurotransmitter metabolism in the mouse brain. Acute administration of chemically modified sericin (0.26 g/30 g body weight) increased L-serine and L-tyrosine levels in the serum and brain, although the L-tyrosine content in the sericin was less than 3% (w/w). In addition, sericin administration led to a significant facilitation of noradrenergic turnover via enhancement of 3-methoxy-4-hydroxyphenylethyleneglycol, a principal metabolite of noradrenaline, in several of the brain regions examined. These present findings suggest that oral intake of sericin efficiently delivers L-serine and L-tyrosine to the brain, thus stimulating noradrenergic activity in the brain.Abbreviations: DA: dopamine; 5-HIAA: 5-hydroxyindoleicetic acid; 5-HT: 5-hydroxytryptamine; HVA: homovanillic acid; MHPG: 3-methoxy-4-hydroxyphenylethyleneglycol; 3-MT: 3-methoxytyramine; NA: noradrenaline; NM: normetanephrine; Veh: vehicle.
Assuntos
Encéfalo/metabolismo , Norepinefrina/metabolismo , Sericinas/administração & dosagem , Serina/metabolismo , Seda/química , Tirosina/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Masculino , Metalotioneína 3 , Camundongos , Camundongos Endogâmicos C57BL , Sericinas/farmacologia , Serina/sangue , Tirosina/sangueRESUMO
The incidence of nonsmoking female patients with non-small cell lung cancer (NSCLC) has increased in recent decades; however, the pathogenesis of patients is unclear, and early diagnosis biomarkers are in urgent need. In this study, 136 nonsmoking female subjects (65 patients with NSCLC, 6 patients with benign lung tumors, and 65 healthy controls) were enrolled, and their metabolic profiling was investigated by using pseudotargeted gas chromatography-mass spectrometry. A total of 56 annotated metabolites were found and verified to be significantly different in nonsmoking females with NSCLC compared with the control. The metabolic profiling was featured by disturbed energy metabolism, amino acid metabolism, oxidative stress, lipid metabolism, and so on. Cysteine, serine, and 1-monooleoylglycerol were defined as the biomarker panel for the diagnosis of NSCLC patients. 98.5 and 91.4% of subjects were correctly distinguished in the discovery and validation sets, respectively. The biomarker panel was also useful for the diagnosis of in situ malignancy patients, with an accuracy of 97.7 and 97.8% in the discovery and validation sets, respectively. The study provides a biomarker panel for the auxiliary diagnosis of nonsmoking females with NSCLC.
Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Cisteína/sangue , Diglicerídeos/sangue , Neoplasias Pulmonares/diagnóstico , Neoplasias/diagnóstico , Serina/sangue , Adulto , Idoso , Aminoácidos/metabolismo , Carcinoma Pulmonar de Células não Pequenas/sangue , Estudos de Casos e Controles , Diagnóstico Diferencial , Detecção Precoce de Câncer/métodos , Metabolismo Energético , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Metabolismo dos Lipídeos , Neoplasias Pulmonares/sangue , Metaboloma , Pessoa de Meia-Idade , Neoplasias/sangue , não Fumantes , Estresse Oxidativo , Sensibilidade e EspecificidadeRESUMO
The aim of the present study was to explore the role of N-methyl-D-aspartate receptor (NMDAR) related amino acids in drug-naive first episode psychosis (FEP) patients. The medication naïve patients with FEP (n = 40) and healthy volunteers with no family history of schizophrenia (n = 35) were recruited to the study and followed up for 10 weeks. Liquid chromatography-mass spectrometry method was used to measure plasma levels of the amino acids. The plasma glutamine, glutamic acid, proline, serine, asparagine, and hydroxyproline levels were significantly higher in the FEP patients compared to healthy controls (p values < .0001). The glutamine/glutamic acid ratio in FEP patients was not different from the healthy controls (p > .05). After the antipsychotic treatment, plasma glutamic acid, proline, and hydroxyproline levels were significantly increased (p values < .05) while the asparagine level and glutamine/glutamic acid ratio were decreased (p values < .05). The serine and glutamine levels did not show any differences with the treatment (p > .05). The initial plasma glutamine levels were negatively correlated with the initial Scale for the Assessment of Positive Symptoms (SAPS) score (r = -.45, p = .003). The initial plasma proline levels were negatively correlated with the initial and follow-up SAPS scores (r = -.51 and -.39, p values < .05). The initial plasma proline and hydroxyproline levels were both negatively correlated with the initial Brief Psychiatric Rating Scale score (r = -.37, p = .017 and r = -.33, p = .033, respectively). Increase in NMDAR-related amino acid levels during the FEP may be a compensatory response to glutamatergic hypofunction. Their plasma levels were significantly correlated with several psychotic symptoms before and after 10-week treatment. Antipsychotic treatment has differential effects on the plasma levels of these amino acids.
Assuntos
Ácido Glutâmico/sangue , Transtornos Psicóticos/sangue , Receptores de N-Metil-D-Aspartato/metabolismo , Esquizofrenia/sangue , Asparagina/sangue , Cromatografia Líquida , Feminino , Glutamina/sangue , Humanos , Hidroxiprolina/sangue , Masculino , Espectrometria de Massas , Prolina/sangue , Serina/sangue , Adulto JovemRESUMO
A novel chiral derivatization reagent, the N-[1-oxo-5-(triphenylphosphonium)pentyl]- (R)-1,3-thiazolidinyl-4-N-hydroxysuccinimide ester bromide salt (OTPTHE), was developed for the separation and selective detection of chiral DL-amino acids by RP-HPLC analysis. The OTPTHE reacted with DL-amino acids at 60°C maintained for 30 minutes in the presence of 100 mM borate buffer (pH 9.5). The separability of the diastereomeric derivatives was evaluated in terms of the resolution value (Rs) using 13 kinds of DL-amino acids, which were completely separated by reversed-phase chromatography using C18 column at 254 nm. The Rs of the DL-amino acids varied from 1.62 to 2.51. As for the application of the DL-amino acids, the determination of DL-Ser in the human plasma of healthy volunteers was performed based on our developed method. It was shown that linear calibrations were available with high coefficients of correlation (r2 > 0.9997). The limit of detection (S/N = 3) of the DL-Ser enantiomers was 5.0 pmol; the relative standard deviations of the intraday and interday variations were below 4.56%; the accuracy ranged between 95.40%-110.06% and 95.45%-109.80%, respectively; the mean recoveries (%) of the DL-Ser spiked in the human plasma were 99.49%-103.74%. The amounts of DL-Ser in the human plasma of healthy volunteers were determined.
Assuntos
Serina/sangue , Serina/química , Succinimidas/química , Tiazolidinas/química , Calibragem , Fracionamento Químico , Cromatografia Líquida , Humanos , Indicadores e Reagentes/química , Serina/isolamento & purificação , EstereoisomerismoRESUMO
Chemical signaling plays a critical role in the behavior and physiology of many animals. Female insects, as many other animals, release sex pheromones to attract males for mating. The evolutionary and ecological success of insects therefore hinges on their ability to precisely mediate (including initiation and termination) pheromone biosynthesis. Pheromone biosynthesis activating neuropeptide (PBAN) acts directly on pheromone glands to regulate sex pheromone production using Ca2+ and cyclic-AMP as secondary messengers in the majority of species. However, the molecular mechanism downstream of the secondary messengers has not yet been elucidated in heliothine species. The present study shows that calcineurin, protein kinase A (PKA) and acetyl-coA carboxylase (ACC) are key components involved in PBAN-induced sex pheromone biosynthesis in Helicoverpa armigera using PBAN-dependent phosphoproteomics in combination with transcriptomics. RNAi-mediated knockdown and inhibitor assay demonstrated that calcineurin A is required for PBAN-induced ACC activation and sex pheromone production. Calcineurin-dependent phosphoproteomics and in vitro calcineurin phosphorylation assay further revealed that calcineurin regulated ACC activity by dephosphorylating ser84 and ser92. In addition, PKA-dependent phosphoproteomics and activity analysis revealed that PKA reduces the activity of AMP-activated protein kinase (AMPK), a negative regulator of ACC by phosphorylating the conserved ser92. Taken together, our findings indicate that calcineurin acts as the downstream signal of PBAN/G-protein receptor/Ca2+ to activate ACC through dephosphorylation while inactivating AMPK via PKA to reduce ACC phosphorylation, thus facilitating calcineurin activation of ACC.
Assuntos
Acetil-CoA Carboxilase/metabolismo , Calcineurina/metabolismo , Perfilação da Expressão Gênica/métodos , Mariposas/metabolismo , Neuropeptídeos/metabolismo , Proteômica/métodos , Acetil-CoA Carboxilase/química , Acetil-CoA Carboxilase/genética , Animais , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Feminino , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Mariposas/genética , Fosforilação , Serina/sangue , Atrativos Sexuais/biossíntese , Espectrometria de Massas em TandemRESUMO
BACKGROUND: 1-Deoxysphingolipids (1-deoxySLs) are atypical sphingolipids. They are formed during sphingolipid de novo synthesis by the enzyme serine palmitoyltransferase, due to the alternate use of alanine over its canonical substrate serine. Pathologically elevated 1-deoxySL are involved in several neurological and metabolic disorders. The objective of this study was to investigate the role of 1-deoxySL in glycogen storage disease type I (GSDI). METHODS: In this prospective, longitudinal observational study (median follow-up 1.8y), the plasma 1-deoxySL profile was analyzed in 15 adult GSDI patients (12 GSDIa, 3 GSDIb), and 31 healthy controls, along with standard parameters for monitoring GSDI. RESULTS: 1-Deoxysphinganine (1-deoxySA) concentrations were elevated in GSDI compared to controls (191⯱â¯129 vs 35⯱â¯14â¯nmol/l, pâ¯<â¯0.0001). Concordant with the mechanism of 1-deoxySL synthesis, plasma alanine was higher (625⯱â¯182 vs 398⯱â¯90⯵mol/l, pâ¯<â¯0.0001), while serine was lower in GSDI than in controls (88⯱â¯22 vs 110⯱â¯18⯵mol/l. pâ¯<â¯0.001). Accordingly, serine, alanine and triglycerides were determinants of 1-deoxySA in the longitudinal analysis of GSDIa. 1-deoxySA concentrations correlated with the occurrence of low blood glucose (area under the curve below 4â¯mmol/l) in continuous glucose monitoring. The 1-deoxySL profile in GSDIb was distinct from GSDIa, with a different ratio of saturated to unsaturated 1-deoxySL. CONCLUSION: In addition to the known abnormalities of lipoproteins, GSDI patients also have a disturbed sphingolipid metabolism with elevated plasma 1-deoxySL concentrations. 1-DeoxySA relates to the occurrence of low blood glucose, and may constitute a potential new biomarker for assessing metabolic control. GSDIa and Ib have distinct 1-deoxySL profiles indicating that both GSD subtypes have diverse phenotypes regarding lipid metabolism.
Assuntos
Doença de Depósito de Glicogênio Tipo I/sangue , Metabolismo dos Lipídeos/genética , Esfingolipídeos/sangue , Adolescente , Adulto , Alanina/sangue , Feminino , Glucose/metabolismo , Doença de Depósito de Glicogênio Tipo I/genética , Doença de Depósito de Glicogênio Tipo I/patologia , Humanos , Masculino , Serina/sangue , Serina C-Palmitoiltransferase/genética , Esfingolipídeos/genética , Adulto JovemRESUMO
Serine biosynthesis defects are autosomal recessive metabolic disorders resulting from the deficiency of any of the three enzymes involved in de novo serine biosynthesis, specifically phosphoglycerate dehydrogenase (PGDH), phosphoserine aminotransferase (PSAT), and phosphoserine phosphatase (PSP). In this study, we performed metabolomic profiling on 4 children with serine biosynthesis defects; 3 with PGDH deficiency and 1 with PSAT deficiency. The evaluations were performed at baseline and with serine and glycine supplementation. Metabolomic profiling performed at baseline showed low phospholipid species, including glycerophosphocholine, glycerophosphoethanolamine, and sphingomyelin. All children had low serine and glycine as expected. Low glycerophosphocholine compounds were found in 4 children, low glycerophosphoethanolamine compounds in 3 children, and low sphingomyelin species in 2 children. Metabolic profiling with serine and glycine supplementation showed normalization of most of the low phospholipid compounds in the 4 children. Phospholipids are the major component of plasma and intracellular membranes, and phosphatidylcholine is the most abundant phospholipid of all mammalian cell types and subcellular organelles. Phosphatidylcholine is of particular importance for the nervous system, where it is essential for neuronal differentiation. The observed low phosphatidylcholine species in children with serine biosynthesis defects that improved after serine supplementation, supports the role of serine as a significant precursor for phosphatidylcholine. The vital role that phosphatidylcholine has during neuronal differentiation and the pronounced neurological manifestations in serine biosynthesis defects suggest that phosphatidylcholine deficiency occurring secondary to serine deficiency may have a significant contribution to the development of the neurological manifestations in individuals with serine biosynthesis defects.
Assuntos
Erros Inatos do Metabolismo dos Carboidratos/metabolismo , Suplementos Nutricionais , Glicina/administração & dosagem , Microcefalia/metabolismo , Fosfatidilcolinas/metabolismo , Fosfoglicerato Desidrogenase/deficiência , Transtornos Psicomotores/metabolismo , Convulsões/metabolismo , Serina/biossíntese , Transaminases/deficiência , Erros Inatos do Metabolismo dos Carboidratos/sangue , Erros Inatos do Metabolismo dos Carboidratos/dietoterapia , Diferenciação Celular , Criança , Pré-Escolar , Feminino , Glicina/sangue , Humanos , Lactente , Masculino , Metabolômica/métodos , Microcefalia/sangue , Microcefalia/dietoterapia , Neurônios/metabolismo , Fosfoglicerato Desidrogenase/sangue , Fosfoglicerato Desidrogenase/metabolismo , Transtornos Psicomotores/sangue , Transtornos Psicomotores/dietoterapia , Convulsões/sangue , Convulsões/dietoterapia , Serina/administração & dosagem , Serina/sangue , Transaminases/sangue , Transaminases/metabolismoRESUMO
To elucidate the molecular mechanisms underlying non-alcoholic fatty liver disease (NAFLD), we recruited 86 subjects with varying degrees of hepatic steatosis (HS). We obtained experimental data on lipoprotein fluxes and used these individual measurements as personalized constraints of a hepatocyte genome-scale metabolic model to investigate metabolic differences in liver, taking into account its interactions with other tissues. Our systems level analysis predicted an altered demand for NAD+ and glutathione (GSH) in subjects with high HS Our analysis and metabolomic measurements showed that plasma levels of glycine, serine, and associated metabolites are negatively correlated with HS, suggesting that these GSH metabolism precursors might be limiting. Quantification of the hepatic expression levels of the associated enzymes further pointed to altered de novo GSH synthesis. To assess the effect of GSH and NAD+ repletion on the development of NAFLD, we added precursors for GSH and NAD+ biosynthesis to the Western diet and demonstrated that supplementation prevents HS in mice. In a proof-of-concept human study, we found improved liver function and decreased HS after supplementation with serine (a precursor to glycine) and hereby propose a strategy for NAFLD treatment.
Assuntos
Glutationa/metabolismo , Lipoproteínas/metabolismo , Metabolômica/métodos , NAD/metabolismo , Hepatopatia Gordurosa não Alcoólica/genética , Serina/administração & dosagem , Animais , Modelos Animais de Doenças , Feminino , Regulação Enzimológica da Expressão Gênica , Genoma , Glicina/sangue , Humanos , Fígado/enzimologia , Fígado/metabolismo , Masculino , Camundongos , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/dietoterapia , Hepatopatia Gordurosa não Alcoólica/metabolismo , Modelagem Computacional Específica para o Paciente , Serina/sangue , Serina/uso terapêuticoRESUMO
Post-traumatic stress disorder (PTSD) is a chronic and debilitating condition that is often refractory to standard frontline antidepressant therapy. A promising new approach to PTSD therapy is administration of a single sub-anesthetic dose of (R,S)-ketamine (Ket). The treatment produces rapid and significant therapeutic response, which lasts for only 4-7â¯days. In one of our studies, the mean duration of response was increased to 33â¯days when Ket administration was combined with a mindfulness-based cognitive therapy, Trauma Interventions using Mindfulness Based Extinction and Reconsolidation (TIMBER). We now report the results from a 20-patient study, which examined the duration of sustained response with combined TIMBER-Ket therapy, TIMBER-K arm, relative to the response observed in a placebo-controlled arm, TIMBER-P. A significant difference in the duration of response was observed between TIMBER-K and TIMBER-P arms: 34.44⯱â¯19.12â¯days and 16.50⯱â¯11.39â¯days, respectively (pâ¯=â¯0.022). Previous studies identified a negative correlation between antidepressant response to Ket and basal plasma concentrations of d-serine (DSR). In this study, the basal DSR levels positively correlated with the pre-treatment severity of PTSD symptoms (Pearson's râ¯=â¯0.42, pâ¯=â¯0.07) and patients with basal DSR levelâ¯≥â¯3.5⯵M displayed not only higher PTSD severity but also shorter duration of response. The data indicate that basal DSR levels may serve as a biomarker of the severity of PTSD symptoms and as a predictor of clinical response. This article is part of a Special Issue entitled: d-Amino acids: biology in the mirror, edited by Dr. Loredano Pollegioni, Dr. Jean-Pierre Mothet and Dr. Molla Gianluca.
Assuntos
Ketamina/uso terapêutico , Atenção Plena , Serina/sangue , Transtornos de Estresse Pós-Traumáticos/terapia , Adulto , Biomarcadores/sangue , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Transtornos de Estresse Pós-Traumáticos/sangueRESUMO
d-Amino acid oxidase (DAAO) is a potential target in the treatment of schizophrenia as its inhibition increases brain d-serine level and thus contributes to NMDA receptor activation. Inhibitors of DAAO were sought testing [6+5] type heterocycles and identified isatin derivatives as micromolar DAAO inhibitors. A pharmacophore and structure-activity relationship analysis of isatins and reported DAAO inhibitors led us to investigate 1H-indazol-3-ol derivatives and nanomolar inhibitors were identified. The series was further characterized by pKa and isothermal titration calorimetry measurements. Representative compounds exhibited beneficial properties in in vitro metabolic stability and PAMPA assays. 6-fluoro-1H-indazol-3-ol (37) significantly increased plasma d-serine level in an in vivo study on mice. These results show that the 1H-indazol-3-ol series represents a novel class of DAAO inhibitors with the potential to develop drug candidates.
Assuntos
D-Aminoácido Oxidase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Indazóis/farmacologia , Animais , D-Aminoácido Oxidase/metabolismo , Relação Dose-Resposta a Droga , Descoberta de Drogas , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Hepatócitos/efeitos dos fármacos , Humanos , Indazóis/síntese química , Indazóis/química , Masculino , Camundongos , Microssomos Hepáticos/efeitos dos fármacos , Modelos Moleculares , Estrutura Molecular , Serina/sangue , Relação Estrutura-AtividadeRESUMO
BACKGROUND: The combination use of the vascular disrupting agent ombrabulin with chemotherapeutic agents was previously shown to be highly synergistic in preclinical models. METHODS: In this dose-escalation study of ombrabulin (15.5-35 mg/m2) in combination with docetaxel (60 or 75 mg/m2) and cisplatin (75 mg/m2), agents were administered 24 h apart every 3 weeks to Japanese patients with advanced solid tumors. The study was designed and conducted in a 3 + 3 manner. Safety, tumor response and pharmacokinetics were evaluated. RESULTS: Eleven patients with non small cell lung cancer as the primary tumor were treated. Two patients out of five had dose limiting toxicities (DLTs) in Cycle 1 at the starting doses of ombrabulin 15.5 mg/m2, docetaxel 60 mg/m2 and cisplatin 75 mg/m2. Thus, dose escalation was terminated. The first dose level was re-evaluated in six patients who received prophylactic granulocyte-colony stimulating factor (G-CSF). However, because of the occurrence of DLTs in Cycle 1 in two patients out of six, the study was led to the premature termination without pursued upper dose level. Partial response was observed in four patients out of 11. Pharmacokinetic parameters of ombrabulin and cisplatin were not altered in this combination treatment, while docetaxel clearance decreased by ~40% compared to that observed with docetaxel monotherapy at the same dose (60 mg/m2). CONCLUSION: A combination regimen of ombrabulin with cisplatin and docetaxel was not feasible for Japanese patients owing to the occurrence of hematological and non-hematological DLTs at the initial dose level. CLINICAL TRIAL REGISTRATION ID: ClinicalTrials.gov number, NCT01095302.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Povo Asiático , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Cisplatino/uso terapêutico , Neoplasias Pulmonares/tratamento farmacológico , Serina/análogos & derivados , Taxoides/uso terapêutico , Adulto , Idoso , Antineoplásicos/efeitos adversos , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/patologia , Cisplatino/administração & dosagem , Cisplatino/efeitos adversos , Cisplatino/farmacocinética , Docetaxel , Relação Dose-Resposta a Droga , Feminino , Humanos , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Serina/efeitos adversos , Serina/sangue , Serina/farmacocinética , Serina/uso terapêutico , Taxoides/efeitos adversos , Taxoides/farmacocinética , Resultado do TratamentoRESUMO
Stroke remains a major public health problem worldwide; it causes severe disability and is associated with high mortality rates. However, early diagnosis of stroke is difficult, and no reliable biomarkers are currently established. In this study, mass-spectrometry-based metabolomics was utilized to characterize the metabolic features of the serum of patients with acute ischemic stroke (AIS) to identify novel sensitive biomarkers for diagnosis and progression. First, global metabolic profiling was performed on a training set of 80 human serum samples (40 cases and 40 controls). The metabolic profiling identified significant alterations in a series of 26 metabolites with related metabolic pathways involving amino acid, fatty acid, phospholipid, and choline metabolism. Subsequently, multiple algorithms were run on a test set consisting of 49 serum samples (26 cases and 23 controls) to develop different classifiers for verifying and evaluating potential biomarkers. Finally, a panel of five differential metabolites, including serine, isoleucine, betaine, PC(5:0/5:0), and LysoPE(18:2), exhibited potential to differentiate AIS samples from healthy control samples, with area under the receiver operating characteristic curve values of 0.988 and 0.971 in the training and test sets, respectively. These findings provided insights for the development of new diagnostic tests and therapeutic approaches for AIS.
Assuntos
Biomarcadores/sangue , Metaboloma/genética , Metabolômica , Acidente Vascular Cerebral/sangue , Idoso , Betaína/sangue , Progressão da Doença , Diagnóstico Precoce , Feminino , Humanos , Isoleucina/sangue , Masculino , Pessoa de Meia-Idade , Curva ROC , Serina/sangue , Acidente Vascular Cerebral/patologiaRESUMO
The objectives of this study were to determine alterations in the serum metabolites related to amino acid (AA), carbohydrate, and lipid metabolism in transition dairy cows before diagnosis of subclinical mastitis (SCM), during, and after diagnosis of disease. A subclinical mastitis case was determined as a cow having somatic cell count (SCC) > 200â¯000/mL of milk for two or more consecutive reports. Blood samples were collected from 100 Holstein dairy cows at five time points at -8 and -4 weeks before parturition, at the week of SCM diagnosis, and +4 and +8 weeks after parturition. Twenty healthy control cows (CON) and six cows that were diagnosed with SCM were selected for serum analysis with GC-MS. At -8 weeks a total of 13 metabolites were significantly altered in SCM cows. In addition, at the week of SCM diagnosis 17 metabolites were altered in these cows. Four weeks after parturition 10 metabolites were altered in SCM cows and at +8 weeks 11 metabolites were found to be different between the two groups. Valine (Val), serine (Ser), tyrosine (Tyr), and phenylalanine (Phe) had very good predictive abilities for SCM and could be used at -8 weeks and -4 weeks before calving. Combination of Val, isoleucine (Ile), Ser, and proline (Pro) can be used as diagnostic biomarkers of SCM during early stages of lactation at +4 to +8 weeks after parturition. In conclusion, SCM is preceded and followed by alteration in AA metabolism.