RESUMO
Purpose: To investigate whether ADSC-derived miR-23-enriched exosomes could protect against calcium oxalate stone formation in a hyperoxaluria rat model. Methods: An ethylene glycol- (EG-) induced hyperoxaluria rat model and an in vitro model of COM-induced HK-2 cells coculturing with RAW264.7 cells were established to explore the protective mechanisms of ADSC-derived miR-23-enriched exosomes. Results: The results showed that treatment with miR-23-enriched exosomes from ADSCs protected EG-induced hyperoxaluria rats, and cell experiments confirmed that coculturing with miR-23-enriched exosomes alleviated COM-induced cell autophagy. Overexpressed miR-23 suppressed M1 macrophage polarization by inhibiting IRF1 expression. Furthermore, the predicted binding site between the IRF1 messenger RNA 3'-untranslated region (3'-UTR) and miR-23 was confirmed by the dual-luciferase reporter assay. Conclusion: In conclusion, our research gave the first evidence that ADSC-derived miR-23-enriched exosomes affected the polarization of M1 macrophages by directly inhibiting IRF1 and protecting against calcium oxalate stone formation in a hyperoxaluria rat model.
Assuntos
Calcinose , Exossomos , Hiperoxalúria , MicroRNAs , Ratos , Animais , Oxalatos , Oxalato de Cálcio/metabolismo , Exossomos/genética , Exossomos/metabolismo , Hiperoxalúria/genética , Hiperoxalúria/metabolismo , Macrófagos/metabolismo , Células Estromais/metabolismo , Calcinose/metabolismo , MicroRNAs/metabolismoRESUMO
The risk of enteric hyperoxaluria is significantly increased after malabsorptive bariatric surgery (MBS). However, its underlying determinants are only poorly characterized. In this case-control study, we aimed at identifying clinical and genetic factors to dissect their individual contributions to the development of post-surgical hyperoxaluria. We determined the prevalence of hyperoxaluria and nephrolithiasis after MBS by 24-h urine samples and clinical questionnaires at our obesity center. Both hyperoxaluric and non-hyperoxaluric patients were screened for sequence variations in known and candidate genes implicated in hyperoxaluria (AGXT, GRHPR, HOGA1, SLC26A1, SLC26A6, SLC26A7) by targeted next generation sequencing (tNGS). The cohort comprised 67 patients, 49 females (73%) and 18 males (27%). While hyperoxaluria was found in 29 patients (43%), only one patient reported postprocedural nephrolithiasis within 41 months of follow-up. Upon tNGS, we did not find a difference regarding the burden of (rare) variants between hyperoxaluric and non-hyperoxaluric patients. However, patients with hyperoxaluria showed significantly greater weight loss accompanied by markers of intestinal malabsorption compared to non-hyperoxaluric controls. While enteric hyperoxaluria is very common after MBS, genetic variation of known hyperoxaluria genes contributes little to its pathogenesis. In contrast, the degree of postsurgical weight loss and levels of malabsorption parameters may allow for predicting the risk of enteric hyperoxaluria and consecutive kidney stone formation.
Assuntos
Cirurgia Bariátrica , Hiperoxalúria , Cálculos Renais , Masculino , Feminino , Humanos , Estudos de Casos e Controles , Hiperoxalúria/genética , Hiperoxalúria/complicações , Cirurgia Bariátrica/efeitos adversos , Cálculos Renais/complicações , Redução de Peso , Variação GenéticaRESUMO
Interaction of pioglitazone (PGZ) and macrophages (Mps) in renal crystal formation remains unclear. In the present study, we investigated the possible mechanisms involved with Mps of PGZ in suppressing renal crystal formation. Crystal formation in the mouse kidney was detected using polarized light optical microscopy and Pizzolato staining. Gene expression was detected by Western blot analysis, quantitative RT-PCR, immunohistochemistry, immunofluorescence, and ELISA. Mp phenotypes were identified by flow cytometric analysis. Cell apoptosis was detected with TUNEL assay, and tubular injury was detected with periodic acid-Schiff staining. Interaction of peroxisome proliferator-activated receptor (PPAR)-γ and promoter was determined by chromatin immunoprecipitation assay. Luciferase reporter assay was performed to authenticate target genes of miRNA-23 (miR-23). Recombinant adenovirus was used to elucidate the role of miR-23 in vivo. Renal crystal formation, inflammation, tubular injury, and cell apoptosis were significantly marked in glyoxylic acid-treated groups and significantly decreased in PGZ-treated groups. PGZ significantly reduced Mp infiltration and M1 Mp polarization in the kidney. In vitro, PGZ shifted crystal-stimulated M1-predominant Mps to M2-predominant Mps, which were anti-inflammatory. PPAR-γ could directly bind to one PPAR-γ regulatory element in the promoter of pre-miR-23 to promote expression of miR-23 in Mps. We identified two downstream target genes of miR-23, interferon regulatory factor 1 and Pknox1. Moreover, miR-23 decreased crystal deposition, M1 Mp polarization, and injury in the kidney. This study has proven that PGZ decreased renal calcium oxalate crystal formation and renal inflammatory injury by suppressing M1 Mp polarization through a PPAR-γ-miR-23-interferon regulatory factor 1/Pknox1 axis. PGZ is liable to be a potential therapeutic medicine for treating urolithiasis.
Assuntos
Anti-Inflamatórios/farmacologia , Oxalato de Cálcio/metabolismo , Hiperoxalúria/prevenção & controle , Rim/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , MicroRNAs/metabolismo , PPAR gama/agonistas , Pioglitazona/farmacologia , Urolitíase/prevenção & controle , Animais , Apoptose/efeitos dos fármacos , Sítios de Ligação , Cristalização , Modelos Animais de Doenças , Regulação da Expressão Gênica , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Hiperoxalúria/genética , Hiperoxalúria/metabolismo , Hiperoxalúria/patologia , Fator Regulador 1 de Interferon/genética , Fator Regulador 1 de Interferon/metabolismo , Rim/metabolismo , Rim/patologia , Macrófagos/metabolismo , Masculino , Camundongos Endogâmicos C57BL , MicroRNAs/genética , PPAR gama/genética , PPAR gama/metabolismo , Fenótipo , Regiões Promotoras Genéticas , Transdução de Sinais , Urolitíase/genética , Urolitíase/metabolismo , Urolitíase/patologiaRESUMO
Most kidney stones (KS) are composed of calcium oxalate, and small increases in urine oxalate affect the stone risk. Intestinal oxalate secretion mediated by anion exchanger SLC26A6 (PAT1) plays a crucial role in limiting net absorption of ingested oxalate, thereby preventing hyperoxaluria and related KS, reflecting the importance of understanding regulation of intestinal oxalate transport. We previously showed that ATP and UTP inhibit oxalate transport by human intestinal Caco2-BBE cells (C2). Since ATP is rapidly degraded to adenosine (ADO), we examined whether intestinal oxalate transport is regulated by ADO. We measured [14C]oxalate uptake in the presence of an outward Cl gradient as an assay of Cl-oxalate exchange activity, ≥49% of which is PAT1-mediated in C2 cells. We found that ADO significantly inhibited oxalate transport by C2 cells, an effect completely blocked by the nonselective ADO receptor antagonist 8- p-sulfophenyltheophylline. ADO also significantly inhibited oxalate efflux by C2 cells, which is important since PAT1 mediates oxalate efflux in vivo. Using pharmacological antagonists and A2B adenosine receptor (A2B AR) siRNA knockdown studies, we observed that ADO inhibits oxalate transport through the A2B AR, phospholipase C, and PKC. ADO inhibits oxalate transport by reducing PAT1 surface expression as shown by biotinylation studies. We conclude that ADO inhibits oxalate transport by lowering PAT1 surface expression in C2 cells through signaling pathways including the A2B AR, PKC, and phospholipase C. Given higher ADO levels and overexpression of the A2B AR in inflammatory bowel disease (IBD), our findings have potential relevance to pathophysiology of IBD-associated hyperoxaluria and related KS.
Assuntos
Adenosina/metabolismo , Sistemas de Transporte de Aminoácidos/genética , Doenças Inflamatórias Intestinais/genética , Receptor A2B de Adenosina/genética , Simportadores/genética , Adenosina/administração & dosagem , Antagonistas do Receptor A2 de Adenosina/administração & dosagem , Trifosfato de Adenosina/metabolismo , Transporte Biológico/genética , Células CACO-2 , Humanos , Hiperoxalúria/genética , Hiperoxalúria/metabolismo , Hiperoxalúria/patologia , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Cálculos Renais/genética , Cálculos Renais/metabolismo , Cálculos Renais/patologia , Oxalatos/metabolismo , Receptor A2B de Adenosina/metabolismo , Fatores de Risco , Transdução de Sinais/efeitos dos fármacos , Teofilina/administração & dosagem , Fosfolipases Tipo C/genéticaRESUMO
Chronic renal failure can give rise to a wide spectrum of oral manifestations, owing mainly to secondary hyperparathyroidism complicating this disease. However, any systemic disease responsible for kidney failure can produce oral manifestations, which can be misdiagnosed. This report describes the case of a 40-year-old male patient referred for oral assessment before kidney and liver transplantation. He had primary hyperoxaluria complicated by end-stage renal failure and secondary hyperparathyroidism. Panoramic radiography indicated not only external root resorption, but also maxillary and mandibular radiolucencies consistent with brown tumors. Unexpectedly, histologic study of the bone biopsy specimen led to the diagnosis of jaws oxalosis. Primary hyperoxaluria is a systemic genetic disease. The affected genes are involved in glyoxylate metabolism and their deficiency results in overproduction of oxalates. Inability of the kidney to excrete oxalates leads to deposition of these crystals in almost all tissues (oxalosis) and to multiple-organ failure. Several oral findings have been described in patients with oxalosis, such as periodontal disease and root resorptions, but radiolucencies in the jaws have rarely been described. This case report is of particular interest because of the unusual location of oxalate crystal deposition in the jaws, which could be misdiagnosed in a patient with renal failure and secondary hyperparathyroidism.
Assuntos
Hiperoxalúria/complicações , Falência Renal Crônica/complicações , Doenças Estomatognáticas/complicações , Adulto , Humanos , Hiperoxalúria/genética , Masculino , Radiografia Panorâmica , Doenças Estomatognáticas/diagnóstico por imagemRESUMO
Osteopontin (OPN) expression is increased in kidneys of rats with ethylene glycol (EG) induced hyperoxaluria and calcium oxalate (CaOx) nephrolithiasis. The aim of this study is to clarify the effect of OPN knockdown by in vivo transfection of OPN siRNA on deposition of CaOx crystals in the kidneys. Hyperoxaluria was induced in 6-week-old male Sprague-Dawley rats by administering 1.5% EG in drinking water for 2 weeks. Four groups of six rats each were studied: Group A, untreated animals (tap water); Group B, administering 1.5% EG; Group C, 1.5% EG with in vivo transfection of OPN siRNA; Group D, 1.5% EG with in vivo transfection of negative control siRNA. OPN siRNA transfections were performed on day 1 and 8 by renal sub-capsular injection. Rats were killed at day 15 and kidneys were removed. Extent of crystal deposition was determined by measuring renal calcium concentrations and counting renal crystal deposits. OPN siRNA transfection resulted in significant reduction in expression of OPN mRNA as well as protein in group C compared to group B. Reduction in OPN expression was associated with significant decrease in crystal deposition in group C compared to group B. Specific suppression of OPN mRNA expression in kidneys of hyperoxaluric rats leads to a decrease in OPN production and simultaneously inhibits renal crystal deposition.
Assuntos
Oxalato de Cálcio/metabolismo , Terapia Genética/métodos , Hiperoxalúria/terapia , Rim/fisiologia , Nefrolitíase/terapia , Osteopontina/genética , Animais , Oxalato de Cálcio/química , Linhagem Celular , Cristalização , Modelos Animais de Doenças , Células Epiteliais/citologia , Hiperoxalúria/genética , Hiperoxalúria/metabolismo , Masculino , Nefrolitíase/genética , Nefrolitíase/metabolismo , Osteopontina/metabolismo , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacologia , Ratos Sprague-DawleyRESUMO
We report a case of anal verruciform xanthoma in a patient who underwent a combined liver and kidney transplantation for primary hyperoxaluria. Verruciform xanthoma is a rare and benign lesion arising in oral cavity and genital mucosa. It is characterized pathologically by papillary epithelial hyperplasia and aggregates of foamy macrophages in connective tissue papillae. This condition, whose pathogenesis remains unclear, has been reported in immunosuppressive background or associated with underlying dermatosis. We report here the second case of anal verruciform xanthoma. To our knowledge, this is the first report of verruciform xanthoma in association with primary hyperoxaluria.
Assuntos
Doenças do Ânus/patologia , Hiperoxalúria/cirurgia , Transplante de Rim , Transplante de Fígado , Complicações Pós-Operatórias/patologia , Xantomatose/patologia , Doenças do Ânus/diagnóstico , Diagnóstico Diferencial , Feminino , Hemorroidas/complicações , Histiócitos/patologia , Humanos , Hiperoxalúria/genética , Hospedeiro Imunocomprometido , Terapia de Imunossupressão/efeitos adversos , Achados Incidentais , Pessoa de Meia-Idade , Plasmócitos/patologia , Reoperação , Verrugas/diagnóstico , Xantogranuloma Juvenil/diagnóstico , Xantomatose/diagnósticoRESUMO
INTRODUCTION: The primary hyperoxalurias (PH) are rare, but underdiagnosed disorders where the loss of enzymatic activity in key compounds of glyoxylate metabolism results in excessive endogenous oxalate generation. Clinically, they are characterized by recurrent urolithiasis and/or nephrocalcinosis. PH type I is the most frequent and most devastating subtype often leading to early end-stage renal failure. AREAS COVERED: Profound overview of clinical, diagnostic, and currently available treatment options with a focus on PH I at different stages of the disease. Discussion of future therapeutic avenues including pharmacological chaperones (small molecules rescuing protein function), gene therapy with safer adenoviral vectors, and potential application of cell-based transplantation strategies is provided. EXPERT OPINION: Due to lack of familiarity with PH and its heterogeneous clinical expression, diagnosis is often delayed until advanced disease is present, a condition, requiring intensive hemodialysis and timely transplantation. Achieving the most beneficial outcome largely depends on the knowledge of the clinical spectrum, early diagnosis, and initiation of treatment before renal failure ensues. A number of preconditions required for substantial improvement in the care of orphan disease like PH have now been achieved or soon will come within reach, so new treatment options can be expected in the near future.
Assuntos
Hiperoxalúria/terapia , Animais , Humanos , Hiperoxalúria/diagnóstico , Hiperoxalúria/genética , Transplante de Rim , Transplante de Fígado , Diálise Renal , Vitamina B 6/uso terapêuticoRESUMO
Oxalate is a metabolic end product excreted by the kidney. Mild increases in urinary oxalate are most commonly associated with Nephrolithiasis. Chronically high levels of urinary oxalate, as seen in patients with primary hyperoxaluria, are driving factor for recurrent renal stones, and ultimately lead to renal failure, calcification of soft tissue and premature death. In previous studies others and we have demonstrated that high levels of oxalate promote injury of renal epithelial cells. However, methods to monitor oxalate induced renal injury are limited. In the present study we evaluated changes in expression of Kidney Injury Molecule-1 (KIM-1) in response to oxalate in human renal cells (HK2 cells) in culture and in renal tissue and urine samples in hyperoxaluric animals which mimic in vitro and in vivo models of hyper-oxaluria. Results presented, herein demonstrate that oxalate exposure resulted in increased expression of KIM-1 m RNA as well as protein in HK2 cells. These effects were rapid and concentration dependent. Using in vivo models of hyperoxaluria we observed elevated expression of KIM-1 in renal tissues of hyperoxaluric rats as compared to normal controls. The increase in KIM-1 was both at protein and mRNA level, suggesting transcriptional activation of KIM-1 in response to oxalate exposure. Interestingly, in addition to increased KIM-1 expression, we observed increased levels of the ectodomain of KIM-1 in urine collected from hyperoxaluric rats. To the best of our knowledge our studies are the first direct demonstration of regulation of KIM-1 in response to oxalate exposure in renal epithelial cells in vitro and in vivo. Our results suggest that detection of KIM-1 over-expression and measurement of the ectodomain of KIM-1 in urine may hold promise as a marker to monitor oxalate nephrotoxicity in hyperoxaluria.
Assuntos
Moléculas de Adesão Celular/metabolismo , Células Epiteliais/metabolismo , Hiperoxalúria/patologia , Rim/patologia , Glicoproteínas de Membrana/metabolismo , Oxalatos/farmacologia , Receptores Virais/metabolismo , Regulação para Cima , Animais , Moléculas de Adesão Celular/química , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/urina , Linhagem Celular , Cristalização , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Receptor Celular 1 do Vírus da Hepatite A , Humanos , Hiperoxalúria/genética , Hiperoxalúria/urina , Imuno-Histoquímica , Rim/efeitos dos fármacos , Rim/metabolismo , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/metabolismo , Túbulos Renais/patologia , Masculino , Glicoproteínas de Membrana/genética , Estrutura Terciária de Proteína , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores Virais/genética , Regulação para Cima/efeitos dos fármacosAssuntos
Hiperoxalúria Primária/diagnóstico , Falência Renal Crônica/etiologia , Nefrocalcinose/etiologia , Transaminases/deficiência , África do Norte/etnologia , Artrite Infecciosa/complicações , Exame de Medula Óssea , Infecções Relacionadas a Cateter/complicações , Terapia Combinada , Descompressão Cirúrgica , Diagnóstico Tardio , Discite/complicações , Discite/cirurgia , Evolução Fatal , Humanos , Hiperoxalúria/complicações , Hiperoxalúria/diagnóstico , Hiperoxalúria/genética , Hiperoxalúria Primária/genética , Hiperoxalúria Primária/patologia , Hipertireoidismo/complicações , Achados Incidentais , Falência Renal Crônica/terapia , Falência Renal Crônica/urina , Laminectomia , Masculino , Diálise Renal , Trombofilia/complicações , Transaminases/genética , Vitamina B 6/uso terapêutico , Adulto JovemRESUMO
It is well known that the incidence of urinary stones is higher in men than women. Although it is believed that the lower incidence of urinary stones in women is due to a protective effect of estrogen, the mechanisms remain unclear. To clarify the relation between female sex hormones and stone matrix protein, we examined the interaction of estrogen receptor-α (ERα), estrogen receptor-related receptor-α (ERRα), and stone matrix protein osteopontin (OPN) in a rat hyperoxaluric model and in primary cultured rat kidney cells. Adult female Wistar rats were divided into 6 groups. Groups 1 and 4 consisted of normal females, Groups 2 and 5 consisted of ovariectomized females, and Groups 3 and 6 consisted of ovariectomized females receiving female sex hormone supplements. Groups 1-3 were administered distilled water, while groups 4-6 were administered 0.5% ethyleneglycol (EG). Moreover, rat kidney primary cultured cells were examined after treatment with female sex hormones under various conditions. The expressions of ERα, ERRα and OPN-mRNA in whole kidney and primary cultured cells were examined using Real-Time PCR. The expressions of OPN and ERRα-mRNA were suppressed by ovariectomy. Supplementation with female sex hormones increased the expression of OPN and ERRα-mRNA. In contrast, the expression of ERα-mRNA was increased by ovariectomy and suppressed by supplementation with female sex hormones. The results of the mRNA expression in primary cultured cells matched those in the hyperoxaluric model rats. Although the reason for the difference in expression between ERα and ERRα-mRNA is unclear, estrogen may regulates OPN expression through ERα and/or ERRα, either independently or in combination. Moreover, the decrease of OPN induced by removal of estrogen may increase urinary stones in postmenopausal women.
Assuntos
Receptor alfa de Estrogênio/genética , Hiperoxalúria/genética , Osteopontina/genética , Receptores de Estrogênio/genética , Animais , Células Cultivadas , Estradiol/análogos & derivados , Estradiol/farmacologia , Feminino , Humanos , Hiperoxalúria/tratamento farmacológico , Hiperoxalúria/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Masculino , Ovariectomia , Progesterona/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptor ERRalfa Relacionado ao EstrogênioRESUMO
Patients with some hereditary nephropathies-including autosomal dominant polycystic kidney disease (ADPKD), Fabry disease and Alport syndrome-can progress to end-stage renal disease (ESRD) and are candidates for kidney transplantation. When considering whether a potential living donor is appropriate for a particular patient, clinicians should be aware of the increased risk of adverse outcomes for the donor and the recipient. Renal transplantation from a living related donor is not contraindicated in most nephropathies that have an autosomal recessive mode of inheritance (for example, autosomal recessive polycystic kidney disease and cystinosis). Renal transplant recipients with ADPKD, however, should only receive a kidney from a related donor if the disease has been excluded in the donor by imaging and/or genetic testing. Potential living related donors for patients with Alport syndrome should be evaluated carefully for the presence of microhematuria and microalbuminuria before a decision is made to perform transplantation, and mothers or heterozygous sisters of affected male recipients with X-linked Alport syndrome should be informed about the possible long-term increased risk of renal dysfunction associated with donation. Most patients with atypical hemolytic uremic syndrome should not receive a kidney transplant from a living donor because there is a high risk of disease recurrence and graft loss.
Assuntos
Nefropatias/genética , Nefropatias/cirurgia , Transplante de Rim , Doadores Vivos , Anormalidades Múltiplas/genética , Anormalidades Múltiplas/cirurgia , Arteriosclerose/genética , Arteriosclerose/cirurgia , Cistinose/genética , Cistinose/cirurgia , Anormalidades do Olho/genética , Anormalidades do Olho/cirurgia , Doença de Fabry/genética , Doença de Fabry/cirurgia , Síndrome Hemolítico-Urêmica/genética , Síndrome Hemolítico-Urêmica/cirurgia , Humanos , Hiperoxalúria/genética , Hiperoxalúria/cirurgia , Síndromes de Imunodeficiência/genética , Síndromes de Imunodeficiência/cirurgia , Doenças Renais Císticas/congênito , Doenças Renais Císticas/genética , Doenças Renais Císticas/cirurgia , Síndromes Miastênicas Congênitas , Nefrite Hereditária/genética , Nefrite Hereditária/cirurgia , Síndrome Nefrótica/genética , Síndrome Nefrótica/cirurgia , Osteocondrodisplasias/genética , Osteocondrodisplasias/cirurgia , Seleção de Pacientes , Rim Policístico Autossômico Dominante/genética , Rim Policístico Autossômico Dominante/cirurgia , Doenças da Imunodeficiência Primária , Embolia Pulmonar/genética , Embolia Pulmonar/cirurgia , Distúrbios Pupilares/genética , Distúrbios Pupilares/cirurgia , Fatores de RiscoRESUMO
Nephrolithiasis associated with inborn metabolic diseases is a very rare condition with some common characteristics: early onset of symptoms, family history, associated tubular impairment, bilateral, multiple and recurrent stones, and association with nephrocalcinosis. The prognosis of such diseases may lead to life threatening conditions, not only because of unabated kidney damage but also because of progressive extra-renal involvement, either in a systemic form (e.g. primary hyperoxaluria type 1, requiring combined liver and kidney transplantation), or in a neurological form (Lesch-Nyhan syndrome leading to auto-mutilation and disability, phosphoribosyl pyrophosphate synthetase superactivity, which is associated with mental retardation). Patients with other inborn metabolic diseases present only with recurrent stone formation, such as cystinuria, adenine phosphoribosyl-transferase deficiency, xanthine deficiency. Finally, nephrolithiasis may be secondarily part of some other metabolic diseases, such as glycogen storage disease type 1 or inborn errors of metabolism leading to Fanconi syndrome (nephropathic cystinosis, tyrosinaemia type 1, fructose intolerance, Wilson disease, respiratory chain disorders, etc.). The diagnosis is based on highly specific investigations, including crystal identification, biochemical analyses and DNA study. The treatment of nephrolithiasis requires hydration as well as specific measures. Compliance is a major issue regarding the progression of renal damage, but the overall outcome mainly depends on extra-renal involvement in relation to the metabolic defect.
Assuntos
Erros Inatos do Metabolismo/complicações , Nefrolitíase/etiologia , Erros Inatos do Metabolismo dos Aminoácidos/complicações , Erros Inatos do Metabolismo dos Aminoácidos/genética , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Criança , Doença de Depósito de Glicogênio Tipo I/complicações , Doença de Depósito de Glicogênio Tipo I/genética , Doença de Depósito de Glicogênio Tipo I/metabolismo , Humanos , Hiperoxalúria/etiologia , Hiperoxalúria/genética , Erros Inatos do Metabolismo/genética , Nefrolitíase/genética , Erros Inatos do Metabolismo da Purina-Pirimidina/complicações , Erros Inatos do Metabolismo da Purina-Pirimidina/genética , Erros Inatos do Metabolismo da Purina-Pirimidina/metabolismoRESUMO
Hereditary monogenic kidney stone diseases are rare diseases, since they account for nearly 2% of nephrolithiasis cases in adults and 10% in children. Most of them are severe, because they frequently are associated with nephrocalcinosis and lead to progressive impairment of renal function unless an early and appropriate etiologic treatment is instituted. Unfortunately, treatment is often lacking or started too late since they are often misdiagnosed or overlooked. The present review reports the genotypic and phenotypic characteristics of monogenic nephrolithiases, with special emphasis on the recent advances in the field of diagnosis and therapeutics. Monogenic stone diseases will be classified into three groups according to their mechanism: (1) inborn errors of the metabolism of oxalate (primary hyperoxalurias), uric acid (hereditary hyperuricemias) or other purines (2,8-dihydroxyadeninuria), which, in addition to stone formation, result in crystal deposition in the renal parenchyma; (2) congenital tubulopathies affecting the convoluted proximal tubule (such as Dent's disease, Lowe syndrome or hypophosphatemic rickets), the thick ascending limb of Henlé's loop (such as familial hypomagnesemia and Bartter's syndromes) or the distal past of the nephron (congenital distal tubular acidosis with or without hearing loss), which are frequently associated with nephrocalcinosis, phosphatic stones and extensive tubulointerstitial fibrosis; (3) cystinuria, an isolated defect in tubular reabsorption of cystine and dibasic aminoacids, which results only in the formation of stones but requires a cumbersome treatment. Analysis of stones appears of crucial value for the early diagnosis of these diseases, as in several of them the morphology and composition of stones is specific. In other cases, especially if nephrocalcinosis, phosphatic stones or proteinuria are present, the evaluation of blood and urine chemistry, especially with regard to calcium, phosphate and magnesium, is the key of diagnosis. Search for mutations is now increasingly performed in as much as genetic counselling is important for the detection of heterozygotes in autosomic recessive diseases and of carrier women in X-linked diseases. In conclusion, better awareness to the rare monogenic forms of nephrolithiasis and/or nephrocalcinosis should allow early diagnosis and treatment which are needed to prevent or substantially delay progression of end-stage renal disease. Analysis of every first stone both in children and in adults should never be neglected, in order to early detect unusual forms of nephrolithiasis requiring laboratory evaluation and deep etiologic treatment.
Assuntos
Cálculos Renais/genética , Humanos , Hiperoxalúria/genética , Hiperoxalúria/terapia , Incidência , Cálculos Renais/diagnóstico , Cálculos Renais/epidemiologia , Cálculos Renais/terapia , Túbulos Renais/patologia , Erros Inatos do Metabolismo/genética , Erros Inatos do Metabolismo/metabolismo , Ácido Oxálico/metabolismoRESUMO
BACKGROUND: Hyperoxaluria may be idiopathic, secondary, or due to primary hyperoxaluria (PH). Hepatic alanine:glyoxylate aminotransferase (AGT) or glyoxylate/hydroxypyruvate reductase (GR/HPR) deficiency causes PHI or PHII, respectively. Hepatic glycolate oxidase (GO) is a candidate enzyme for a third form of inherited hyperoxaluria. METHODS: Six children were identified with marked hyperoxaluria, urolithiasis, and normal hepatic AGT (N = 5) and GR/HPR (N = 4). HPR was below normal and GR not measured in one. Of an affected sibling pair, only one underwent biopsy. GO mutation screening was performed, and dietary oxalate (Diet(ox)), enteric oxalate absorption (EOA) measured using [13C2] oxalate, renal clearance (GFR), fractional oxalate excretion (FE(ox)) in the children, and urine oxalate in first-degree relatives (FDR) to understand the etiology of the hyperoxaluria. RESULTS: Mean presenting age was 19.2 months and urine oxalate 1.3 +/- 0.5 mmol/1.73 m2/24 h (mean +/- SD). Two GO sequence changes (T754C, IVS3 - 49 C>G) were detected which were not linked to the hyperoxaluria. Diet(ox) was 42 +/- 31 mg/day. EOA was 9.4 +/- 3.6%, compared with 7.6 +/- 1.2% in age-matched controls (P = 0.33). GFR was 90 +/- 19 mL/min/1.73 m2 and FE(ox) 4.2 +/- 1.4. Aside from the two brothers, hyperoxaluria was not found in FDR. CONCLUSIONS: These patients illustrate a novel form of hyperoxaluria and urolithiasis, without excess Diet(ox), enteric hyper-absorption, or hepatic AGT, GR/HPR deficiency. Alterations in pathways of oxalate synthesis, in liver or kidney, or in renal tubular oxalate handling are possible explanations. The affected sibling pair suggests an inherited basis.
Assuntos
Hiperoxalúria/genética , Hiperoxalúria/metabolismo , Oxirredutases do Álcool/genética , Cálcio da Dieta/urina , Pré-Escolar , Saúde da Família , Feminino , Glioxilatos/metabolismo , Humanos , Hiperoxalúria Primária , Lactente , Masculino , Dados de Sequência Molecular , Oxalatos/urina , Linhagem , Fosfatos/urina , Cálculos Urinários/genética , Cálculos Urinários/metabolismoRESUMO
BACKGROUND AND PURPOSE: The primary hyperoxalurias are a group of inherited disorders of endogenous oxalate overproduction. Diagnosis of the two best-characterized disorders, primary hyperoxaluria (PH) Types 1 and 2, is achieved by sequential measurement of alanine:glyoxylate aminotransferase and glyoxylate reductase enzyme activity in a single needle liver biopsy. While genetic analysis of PH2 is still at a relatively early stage, the AGXT gene defective in the Type 1 disorder is well characterized, and a number of mutations have been identified. METHODS: To determine whether mutation analysis could replace enzymatic analysis for the diagnosis of PH1, DNA samples from 127 consecutive unrelated patients in whom there was a high clinical suspicion of primary hyperoxaluria were analyzed for the presence of the G630A and T853C mutations, which together account for approximately 34% of the mutant alleles in our patient cohort. RESULTS AND CONCLUSIONS: The sensitivity of mutation detection was 47% in those patients with enzymologically confirmed Type 1 disease, showing that mutation analysis cannot effectively replace enzymology at the present time. However, there is little doubt of the value of genetic methods (mutation and linkage analysis) for diagnosing PH1 (and eventually PH2) in other family members and for prenatal diagnosis and carrier testing.
Assuntos
Hiperoxalúria/diagnóstico , Hiperoxalúria/genética , Oxirredutases do Álcool/metabolismo , Alelos , Biópsia , Saúde da Família , Feminino , Ligação Genética , Marcadores Genéticos , Humanos , Hidroxipiruvato Redutase , Hiperoxalúria Primária/diagnóstico , Hiperoxalúria Primária/genética , Íntrons , Fígado/enzimologia , Masculino , Repetições de Microssatélites , Mutação , Linhagem , Diagnóstico Pré-Natal , Recombinação Genética , Sensibilidade e Especificidade , Transaminases/genética , Transaminases/metabolismoRESUMO
Considering the clinical heterogeneity of primary hyperoxaluria type I (PH1) and the fact that in many instances this diagnosis was made without enzymatic and immunohistochemical investigation, other disturbances of oxalate metabolism than those presently known can be expected in PH1. Using a gaschromatographic/mass spectrometric method that allows quantification of these acids, hyperoxaluria and hyperglycoluria was found repeatedly in two unrelated patients. The hyperoxaluria was unresponsive to pyridoxine. There was no nephrocalcinosis or urolithiasis. In the liver biopsy normal AGT activity and normal localization of this enzyme in the peroxisome was found. In one patient abnormal Km and maximal activity and mozaicism of AGT were excluded. Hyperoxaluria and hyperglycoluria were also found in other family members, suggesting autosomal dominant transmission. Although the underlying defect leading to hyperoxaluria and hyperglycoluria could not be identified in these patients, it is probable that they represent a separate type of primary hyperoxaluria.
Assuntos
Alanina Transaminase/metabolismo , Glicosúria Renal/etiologia , Hiperoxalúria/etiologia , Transaminases , Alanina Transaminase/genética , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Glicolatos/urina , Glicosúria Renal/genética , Humanos , Hiperoxalúria/genética , Imuno-Histoquímica , Lactente , Cinética , Fígado/patologia , Fígado/ultraestrutura , Masculino , Microcorpos/enzimologiaRESUMO
The autosomal recessive disease primary hyperoxaluria type 1 (PH1) is caused by a deficiency of the liver-specific peroxisomal enzyme alanine:glyoxylate amino-transferase (AGT). This paper concerns the identification, characterization and clinical use of an unusual discretely polymorphic tandem repeat sequence in the fourth intron of the human AGT gene (gene locus designation AGXT). In a random Caucasian population, three alleles could be clearly recognized that consisted of either 12 (type III), 17 (type II) or approximately 38 (type I) tandemly repeated copies of a highly conserved 29/32-bp sequence with frequencies of 33%, 7% and 60%, respectively. In a random Japanese population, the allelic frequencies were markedly different (i.e. 31%, 45% and 19%, respectively). In addition, a fourth allele was identified, consisting of approximately 32 repeats (type IV), with an allelic frequency of approximately 5% in Japanese. The repetitive sequence was similar to previously identified mammalian sequences with homology to the Epstein-Barr virus IR3 repetitive element involving a 12/15-bp region GCA(GGN)GGAGGAGGG within the repeat unit. This IR3-like sequence was interspersed with a 17-bp sequence with no similarity to any currently known repetitive element. The type I and type III alleles were judged to be equivalent to a previously identified TaqI polymorphism. Two polymorphisms previously shown to be associated with the peroxisome-to-mitochondrion mistargeting of AGT in PH1 (a C154-->T point substitution in exon 1 and a 74-bp duplication in intron 1) were found to segregate exclusively with the type I intron 4 polymorphism in Caucasians, but not in Japanese. The polymorphic nature of the intron 4 tandem repeats makes them of potential use in the prenatal diagnosis of PH1, especially when coupled with the exon 1 C154-->T substitution or intron 1 duplication polymorphisms. A PH1 family, in which a fetus had been predicted previously to be either normal or a carrier by AGT enzymic analysis of a fetal liver biopsy, but who had been shown to be only partially informative with respect to the C154-->T/intron 1 polymorphisms, was analysed retrospectively. The family was completely informative for the intron 4 tandem repeat polymorphism and the carrier status of the fetus was confirmed.
Assuntos
Alanina Transaminase/genética , Hiperoxalúria/diagnóstico , Íntrons , Polimorfismo Genético , Sequências Repetitivas de Ácido Nucleico , Transaminases , Alanina Transaminase/deficiência , Alelos , Sequência de Bases , DNA , Feminino , Frequência do Gene , Humanos , Hiperoxalúria/genética , Fígado/enzimologia , Masculino , Dados de Sequência Molecular , Linhagem , Diagnóstico Pré-Natal , Estudos RetrospectivosRESUMO
In approximately one-third of primary hyperoxaluria type 1 patients, disease is associated with a unique protein sorting defect in which hepatic L-alanine:glyoxylate aminotransferase (AGT; EC 2.6.1.44), which is normally peroxisomal, is mistargeted to mitochondria. In all such patients analyzed to date, the gene encoding the aberrantly targeted AGT carries three point mutations, each of which specifies an amino acid substitution. In this paper we show that one of these substitutions, a proline-to-leucine at residue 11, is necessary and sufficient for the generation of a mitochondrial targeting sequence in the AGT protein. AGT with this substitution appears to interact specifically with the mitochondrial protein import machinery, via a discrete N-terminal domain of the AGT protein. The N-terminal 19 amino acids of AGT with this substitution are sufficient to direct mouse cytosolic dihydrofolate reductase to mitochondria, and a synthetic peptide corresponding to this same 19-amino acid region reversibly inhibits mitochondrial protein import, not only of AGT but also of ornithine transcarbamoylase, a genuine cytoplasmically synthesized mitochondrial protein. We have extended these studies to analyze a region of normal human AGT cDNA directly upstream of the coding region. This sequence appears to correspond to an ancestral mitochondrial targeting sequence deleted from the human coding region by point mutation at the initiation codon. We show that reestablishment of this initiation codon produces an active mitochondrial targeting sequence that is different to that found in the hyperoxaluria patients. These results are discussed with reference to the AGT targeting defect in primary hyperoxaluria and also in relation to the highly unusual species specificity of subcellular distribution of AGT among mammals.