ABSTRACT
Gitelman and Bartter syndromes are rare inherited diseases that belong to the category of renal tubulopathies. The genes associated with these pathologies encode electrolyte transport proteins located in the nephron, particularly in the Distal Convoluted Tubule and Ascending Loop of Henle. Therefore, both syndromes are characterized by alterations in the secretion and reabsorption processes that occur in these regions. Patients suffer from deficiencies in the concentration of electrolytes in the blood and urine, which leads to different systemic consequences related to these salt-wasting processes. The main clinical features of both syndromes are hypokalemia, hypochloremia, metabolic alkalosis, hyperreninemia and hyperaldosteronism. Despite having a different molecular etiology, Gitelman and Bartter syndromes share a relevant number of clinical symptoms, and they have similar therapeutic approaches. The main basis of their treatment consists of electrolytes supplements accompanied by dietary changes. Specifically for Bartter syndrome, the use of non-steroidal anti-inflammatory drugs is also strongly supported. This review aims to address the latest diagnostic challenges and therapeutic approaches, as well as relevant recent research on the biology of the proteins involved in disease. Finally, we highlight several objectives to continue advancing in the characterization of both etiologies.
Subject(s)
Bartter Syndrome/pathology , Gitelman Syndrome/pathology , Kidney Tubules, Distal/pathology , Loop of Henle/pathology , Water-Electrolyte Balance/physiology , Bartter Syndrome/diagnosis , Bartter Syndrome/genetics , Bartter Syndrome/therapy , Electrolytes/analysis , Electrolytes/therapeutic use , Gitelman Syndrome/diagnosis , Gitelman Syndrome/genetics , Gitelman Syndrome/therapy , Humans , Hyperaldosteronism/pathology , Hypercalciuria/pathology , Hypokalemia/pathology , Hyponatremia/pathology , Nephrocalcinosis/pathology , Renal Tubular Transport, Inborn Errors/pathologySubject(s)
Hypercalciuria/blood , Hypercalciuria/diagnosis , Laboratories, Hospital , Magnesium Deficiency/blood , Magnesium/blood , Nephrocalcinosis/blood , Nephrocalcinosis/diagnosis , Renal Tubular Transport, Inborn Errors/blood , Renal Tubular Transport, Inborn Errors/diagnosis , Emergency Service, Hospital , Humans , Hypercalciuria/pathology , Nephrocalcinosis/pathology , Renal Tubular Transport, Inborn Errors/pathology , Undiagnosed DiseasesABSTRACT
Urinary calcium and magnesium wasting is a characteristic feature of metabolic acidosis, and this study focused on the role of the thick ascending limb of Henle's loop in metabolic acidosis-induced hypercalciuria and hypermagnesiuria because thick ascending limb is an important site of paracellular calcium and magnesium reabsorption. Male Sprague-Dawley rats were used to determine the effects of acid loading (by adding NH4Cl, 7.2 mmol/220 g body wt/day to food slurry for 7 days) on renal expression of claudins and then to evaluate whether the results were reversed by antagonizing calcium-sensing receptor (using NPS-2143). At the end of each animal experiment, the kidneys were harvested for immunoblotting, immunofluorescence microscopy, and quantitative PCR (qPCR) analysis of claudins and the calcium-sensing receptor. As expected, NH4Cl loading lowered urinary pH and increased excretion of urinary calcium and magnesium. In NH4Cl-loaded rats, renal protein and mRNA expression of claudin-16, and claudin-19, were decreased compared with controls. However, claudin-14 protein and mRNA increased in NH4Cl-loaded rats. Consistently, the calcium-sensing receptor protein and mRNA were up-regulated in NH4Cl-loaded rats. All these changes were reversed by NPS-2143 coadministration and were confirmed using immunofluorescence microscopy. Hypercalciuria and hypermagnesiuria in NH4Cl-loaded rats were significantly ameliorated by NPS-2143 coadministration as well. We conclude that in metabolic acidosis, claudin-16 and claudin-19 in the thick ascending limb are down-regulated to produce hypercalciuria and hypermagnesiuria via the calcium-sensing receptor.NEW & NOTEWORTHY This study found that the thick ascending limb of Henle's loop is involved in the mechanisms of hypercalciuria and hypermagnesiuria in metabolic acidosis. Specifically, expression of claudin-16/19 and claudin-14 was altered via up-regulation of calcium-sensing receptor in NH4Cl-induced metabolic acidosis. Our novel findings contribute to understanding the regulatory role of paracellular tight junction proteins in the thick ascending limb.
Subject(s)
Calcium/metabolism , Claudins/metabolism , Hypercalciuria/metabolism , Loop of Henle/metabolism , Magnesium/metabolism , Acidosis/metabolism , Animals , Calcium, Dietary/metabolism , Hypercalciuria/pathology , Loop of Henle/pathology , Male , Rats, Sprague-Dawley , Receptors, Calcium-Sensing/metabolismABSTRACT
CONTEXT: The calcium-sensing receptor (CaSR) is essential to maintain a stable calcium concentration in serum. Spermatozoa are exposed to immense changes in concentrations of CaSR ligands such as calcium, magnesium, and spermine during epididymal maturation, in the ejaculate, and in the female reproductive environment. However, the role of CaSR in human spermatozoa is unknown. OBJECTIVE: This work aimed to investigate the role of CaSR in human spermatozoa. METHODS: We identified CaSR in human spermatozoa and characterized the response to CaSR agonists on intracellular calcium, acrosome reaction, and 3',5'-cyclic adenosine 5'-monophosphate (cAMP) in spermatozoa from men with either loss-of-function or gain-of-function mutations in CASR and healthy donors. RESULTS: CaSR is expressed in human spermatozoa and is essential for sensing extracellular free ionized calcium (Ca2+) and Mg2+. Activators of CaSR augmented the effect of sperm-activating signals such as the response to HCO3- and the acrosome reaction, whereas spermatozoa from men with a loss-of-function mutation in CASR had a diminished response to HCO3-, lower progesterone-mediated calcium influx, and were less likely to undergo the acrosome reaction in response to progesterone or Ca2+. CaSR activation increased cAMP through soluble adenylyl cyclase (sAC) activity and increased calcium influx through CatSper. Moreover, external Ca2+ or Mg2+ was indispensable for HCO3- activation of sAC. Two male patients with a CASR loss-of-function mutation in exon 3 presented with normal sperm counts and motility, whereas a patient with a loss-of-function mutation in exon 7 had low sperm count, motility, and morphology. CONCLUSION: CaSR is important for the sensing of Ca2+, Mg2+, and HCO3- in spermatozoa, and loss-of-function may impair male sperm function.
Subject(s)
Bicarbonates/metabolism , Calcium/metabolism , Receptors, Calcium-Sensing/physiology , Spermatozoa/metabolism , Acrosome Reaction/drug effects , Acrosome Reaction/genetics , Adult , Bicarbonates/pharmacology , Calcium/pharmacology , Calcium Signaling/drug effects , Calcium Signaling/genetics , Case-Control Studies , Female , Humans , Hypercalcemia/congenital , Hypercalcemia/genetics , Hypercalcemia/metabolism , Hypercalcemia/pathology , Hypercalciuria/genetics , Hypercalciuria/metabolism , Hypercalciuria/pathology , Hypocalcemia/genetics , Hypocalcemia/metabolism , Hypocalcemia/pathology , Hypoparathyroidism/congenital , Hypoparathyroidism/genetics , Hypoparathyroidism/metabolism , Hypoparathyroidism/pathology , Kidney/metabolism , Kidney/pathology , Magnesium/metabolism , Magnesium/pharmacology , Male , Mutation , Receptors, Calcium-Sensing/genetics , Sperm Motility/drug effects , Sperm Motility/genetics , Spermatozoa/drug effects , Spermatozoa/physiology , Testicular Neoplasms/genetics , Testicular Neoplasms/metabolism , Testicular Neoplasms/pathologyABSTRACT
CONTEXT: Human cytochrome P450 24 subfamily A member 1 (CYP24A1) loss-of-function mutations result in impaired activity of the 24-hydroxylase involved in vitamin D catabolism, thus inducing a vitamin D-dependent hypercalcemia. Homozygotes often present an overt clinical phenotype named idiopathic infantile hypercalcemia (IIH), whereas it is debated whether heterozygotes display an abnormal phenotype. OBJECTIVE: To compare the clinical and biochemical features of heterozygous carriers of CYP24A1 variant and healthy wild-type controls sharing the same genetic and environmental exposure. METHODS: A large family harboring the nonsense c.667A>T, p.Arg223* pathogenic variant in the CYP24A1 gene was evaluated. All subjects underwent clinical and biochemical evaluation and complete analysis of vitamin D metabolites using mass spectroscopy including 1,24,25(OH)3D3. Subjects were divided into 2 groups according to their genotype: heterozygotes and wild-type for the CYP24A1 variant. RESULTS: The proband, a 40-year-old man, homozygous for p.Arg223* pathogenic variant, had a history of mild hypercalcemia with a seasonal trend, recurrent nephrolithiasis, and no episodes of acute hypercalcemia. He showed the highest serum levels of fibroblast growth factor 23, the highest 25(OH)D3/24,25(OH)2D3 ratio and undetectable levels of 1,24,25(OH)3D3, which represent indicators of a loss-of-function CYP24A1. Compared with the wild-types, heterozygotes had higher serum calcium and 25(OH)D3 concentrations (P = .017 and P = .025, respectively), without any difference in the other biochemical parameters and in the rate of nephrolithiasis. CONCLUSION: Heterozygotes exhibit a biochemical phenotype different from that of wild-type subjects. In clinical practice, these individuals might require surveillance because of the potential risk of developing hypercalcemia and related clinical manifestations if exposed to triggering factors.
Subject(s)
Hypercalcemia/blood , Hypercalcemia/genetics , Vitamin D3 24-Hydroxylase/genetics , Adult , Biological Variation, Population , Biomarkers/blood , Case-Control Studies , Codon, Nonsense , Family , Fibroblast Growth Factor-23 , Fibroblast Growth Factors/blood , Genotype , Heterozygote , Homozygote , Humans , Hypercalcemia/pathology , Hypercalciuria/blood , Hypercalciuria/genetics , Hypercalciuria/pathology , Male , Nephrocalcinosis/blood , Nephrocalcinosis/genetics , Nephrocalcinosis/pathology , Pedigree , Phenotype , Vitamin D/bloodABSTRACT
Hypercalcemia is a rare presentation of childhood acute lymphoblastic leukemia (ALL), and presents with nonspecific symptoms. A 11-year old boy developed severe hypercalcemia during initial presentation and relapse of ALL. Both times, he subsequently developed transient symptomatic hypocalcemia, associated with hypomagnesemia and renal tubulopathy. Disturbances in calcium homeostasis may rarely be the sole presenting feature of ALL in children, as a paraneoplastic syndrome, or may arise as a consequence of the malignancy and its treatment. Along with other measures, early recognition of malignancy and initiation of treatment play a key role in correcting calcium disturbances.
Subject(s)
Hypercalciuria/pathology , Hypocalcemia/pathology , Kidney Diseases/pathology , Nephrocalcinosis/pathology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/complications , Renal Tubular Transport, Inborn Errors/pathology , Child , Homeostasis , Humans , Hypercalciuria/etiology , Hypocalcemia/etiology , Kidney Diseases/etiology , Male , Nephrocalcinosis/etiology , Prognosis , Renal Tubular Transport, Inborn Errors/etiologyABSTRACT
PURPOSE: Cetuximab and panitumumab are monoclonal antibodies that target the epidermal growth factor receptor (EGFR). Treatment with cetuximab and panitumumab commonly causes hypomagnesemia, and optimal management of this adverse effect remains unclear. Here, we evaluated the optimal magnesium replacement points based on the risk of severe hypomagnesemia in colorectal cancer patients who received cetuximab or panitumumab. METHODS: We retrospectively evaluated 184 patients who received cetuximab or panitumumab for colorectal cancer at Ogaki Municipal Hospital (Ogaki, Japan) between January 2010 and December 2019. Univariate analyses were conducted to evaluate the relationship between patient baseline characteristics and development of hypomagnesemia following cetuximab or panitumumab treatment. Variables that were significantly associated with hypomagnesemia in the univariate analyses as well as previously reported risk factors were entered into a multivariate logistic regression model. RESULTS: The incidence of hypomagnesemia was associated with panitumumab treatment, pre-replenishment serum magnesium concentration, treatment duration, and treatment line. Severe hypomagnesemia post-cetuximab or panitumumab treatment was significantly associated with low baseline magnesium concentrations (< 1.8 mg/dL; odds ratio 18.100, 95% confidence interval 1.570-210.000; p = 0.020) and low serum magnesium concentrations during treatment (< 1.1 mg/dL; odds ratio 93.800, 95% confidence interval 3.510-2510.000; p = 0.007). CONCLUSION: To minimize the risk of severe hypomagnesemia during anti-EGFR treatment, magnesium replenishment should be initiated in patients with pre-replenishment concentrations of < 1.8 mg/dL, preferably before reaching intra-treatment concentrations of < 1.1 mg/dL.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Colorectal Neoplasms/drug therapy , Hypercalciuria/prevention & control , Magnesium/administration & dosage , Neoplasm Recurrence, Local/drug therapy , Nephrocalcinosis/prevention & control , Renal Tubular Transport, Inborn Errors/prevention & control , Adult , Aged , Aged, 80 and over , Cetuximab/administration & dosage , Colorectal Neoplasms/pathology , Female , Follow-Up Studies , Humans , Hypercalciuria/chemically induced , Hypercalciuria/pathology , Male , Middle Aged , Neoplasm Recurrence, Local/pathology , Nephrocalcinosis/chemically induced , Nephrocalcinosis/pathology , Panitumumab/administration & dosage , Prognosis , Renal Tubular Transport, Inborn Errors/chemically induced , Renal Tubular Transport, Inborn Errors/pathology , Retrospective StudiesABSTRACT
BACKGROUND: Barakat syndrome is an autosomal dominant disorder characterized by the triad of hypoparathyroidism, sensorineural deafness, and renal anomalies and is caused by mutations in GATA3 gene. SLC34A3 is the cause gene of hypophosphatemic rickets with hypercalciuria, and heterozygous carriers may have milder clinical symptoms. The aim of this study was to identify the underlying genetic cause of a patient who initially presented with renal failure, hypercalciuria, kidney stone, and bilateral sensorineural deafness. METHODS: A 6-year-old boy with complex clinical presentations was investigated. Comprehensive medical evaluations were performed including auditory function tests, endocrine function tests, metabolic studies, and imaging examinations. Molecular diagnoses were analyzed by trio whole-exome sequencing. RESULTS: One novel de novo deleterious variant (c. 324del) of the GATA3 gene was identified in the patient. The patient can be diagnosed with Barakat syndrome. In addition, one novel variant (c. 589A>G) of the SLC34A3 gene was detected, which was inherited from the father. This heterozygous variant can explain the hypercalciuria and kidney stone that occurred in both the patient and his father. CONCLUSION: This study provides a special case which is phenotype-driven dual diagnoses, and the two novel variants can parsimoniously explain the complex clinical presentations of this patient.
Subject(s)
GATA3 Transcription Factor/genetics , Hearing Loss, Sensorineural/genetics , Hypercalciuria/genetics , Hypoparathyroidism/genetics , Mutation , Nephrosis/genetics , Sodium-Phosphate Cotransporter Proteins, Type IIc/genetics , Adult , Child , Hearing Loss, Sensorineural/complications , Hearing Loss, Sensorineural/pathology , Heterozygote , Humans , Hypercalciuria/complications , Hypercalciuria/pathology , Hypoparathyroidism/complications , Hypoparathyroidism/pathology , Male , Nephrosis/complications , Nephrosis/pathology , PedigreeABSTRACT
The major risk factor for kidney stone disease is idiopathic hypercalciuria. Recent evidence implicates a role for defective calcium reabsorption in the renal proximal tubule. We hypothesized that claudin-2, a paracellular cation channel protein, mediates proximal tubule calcium reabsorption. We found that claudin-2-null mice have hypercalciuria due to a primary defect in renal tubule calcium transport and papillary nephrocalcinosis that resembles the intratubular plugs in kidney stone formers. Our findings suggest that a proximal tubule defect in calcium reabsorption predisposes to papillary calcification, providing support for the vas washdown hypothesis. Claudin-2-null mice were also found to have increased net intestinal calcium absorption, but reduced paracellular calcium permeability in the colon, suggesting that this was due to reduced intestinal calcium secretion. Common genetic variants in the claudin-2 gene were associated with decreased tissue expression of claudin-2 and increased risk of kidney stones in 2 large population-based studies. Finally, we describe a family in which males with a rare missense variant in claudin-2 have marked hypercalciuria and kidney stone disease. Our findings indicate that claudin-2 is a key regulator of calcium excretion and a potential target for therapies to prevent kidney stones.
Subject(s)
Claudins , Gene Expression Regulation , Genetic Variation , Hypercalciuria , Kidney Calculi , Kidney Tubules, Proximal , Animals , Calcium/urine , Claudins/deficiency , Claudins/metabolism , Hypercalciuria/genetics , Hypercalciuria/pathology , Hypercalciuria/urine , Kidney Calculi/genetics , Kidney Calculi/pathology , Kidney Calculi/urine , Kidney Tubules, Proximal/metabolism , Kidney Tubules, Proximal/pathology , Mice , Mice, KnockoutABSTRACT
ABSTRACT Sarcoidosis is a multisystem granulomatous disease characterized by epithelioid noncaseating granulomas associated with clinical and radiologic findings. The cause of this disease is still uncertain. Sarcoidosis affects mostly lungs and lymph nodes and is not usually considered a urological disease, therefore, this etiology may be overlooked in several urological disorders, such as hypercalcemia, hypercalciuria and nephrolithiasis. It affects all races and genders. This review aims to describe the urological manifestations of sarcoidosis and to elucidate how the disease may affect the management of numerous urological conditions.
Subject(s)
Humans , Sarcoidosis/pathology , Kidney Diseases/pathology , Sarcoidosis/diagnosis , Sarcoidosis/therapy , Biopsy , Chronic Disease , Hypercalciuria/diagnosis , Hypercalciuria/pathology , Hypercalcemia/diagnosis , Hypercalcemia/pathology , Kidney Diseases/diagnosis , Kidney Diseases/therapyABSTRACT
Sarcoidosis is a multisystem granulomatous disease characterized by epithelioid noncaseating granulomas associated with clinical and radiologic findings. The cause of this disease is still uncertain. Sarcoidosis affects mostly lungs and lymph nodes and is not usually considered a urological disease, therefore, this etiology may be overlooked in several urological disorders, such as hypercalcemia, hypercalciuria and nephrolithiasis. It affects all races and genders. This review aims to describe the urological manifestations of sarcoidosis and to elucidate how the disease may affect the management of numerous urological conditions.
Subject(s)
Kidney Diseases/pathology , Sarcoidosis/pathology , Biopsy , Chronic Disease , Humans , Hypercalcemia/diagnosis , Hypercalcemia/pathology , Hypercalciuria/diagnosis , Hypercalciuria/pathology , Kidney Diseases/diagnosis , Kidney Diseases/therapy , Sarcoidosis/diagnosis , Sarcoidosis/therapyABSTRACT
The expression of vitamin D receptor (VDR) and 1,25-dihydroxyvitamin D3 [1,25(OH)D] levels exceed the values of controls in some but not all hypercalciuric stone formers (HSF). We aimed to evaluate serum 1,25(OH)D levels, the expression of VDR, CYP27B1, and CYP24A1 hydroxylases in HSF in comparison with normocalciuric stone formers (NSF) and healthy subjects (HS). Blood samples, 24-h urine collections and a 3-day dietary record were obtained from 30 participants from each of the groups. The expression of VDR, CYP27B1, and CYP24A1 was measured by flow cytometry. HSF presented significantly higher urinary volume, sodium, magnesium, oxalate, uric acid, and phosphorus than NSF and HS. Calcium intake was lower in HSF versus NSF and HS (442 ± 41 vs 594 ± 42 and 559 ± 41 mg/day, respectively, p = 0.027). Ionized calcium was significantly lower in HSF than NSF (1.29 ± 0.0 vs 1.31 ± 0.0 mmol/L, p < 0.01). Serum 1,25(OH)D was significantly higher in HSF and NSF than HS (22.5 ± 1.2; 22.2 ± 1.2 vs 17.4 ± 1.2 pg/ml, p = 0.007) but serum 25(OH)D, PTH, klotho and plasma FGF-23 did not differ between groups. VDR expression was higher in HSF and NSF than HS (80.8 ± 3.2; 78.7 ± 3.3 vs 68.6 ± 3.2%, p = 0.023). Although CYP27B1 and CYP24A1 expressions were similar among all groups, the ratio of 1,25(OH)D/CYP24A1 was higher in HSF and NSF than in HS (1.43 ± 0.25 and 0.56 ± 0.10 vs 0.34 ± 0.06, p = 0.00). Stone formers, regardless of urinary calcium excretion, had higher VDR expression and 1,25(OH)D levels than HS, even in ranges considered normal. Higher 1,25(OH)D/CYP24A1 ratio suggested a lower degradation of 1,25(OH)D by CYP24A1 in HSF and NSF.
Subject(s)
25-Hydroxyvitamin D3 1-alpha-Hydroxylase/metabolism , Calcitriol/blood , Hypercalciuria/pathology , Kidney Calculi/pathology , Receptors, Calcitriol/metabolism , Vitamin D3 24-Hydroxylase/metabolism , Adult , Calcitriol/metabolism , Calcium/urine , Fibroblast Growth Factor-23 , Healthy Volunteers , Humans , Hypercalciuria/blood , Hypercalciuria/complications , Hypercalciuria/urine , Kidney Calculi/blood , Kidney Calculi/etiology , Kidney Calculi/urine , Leukocytes, Mononuclear/metabolism , MaleABSTRACT
People with calcium oxalate (CaOx) urolithiasis and idiopathic hypercalciuria (IH) often have evidence of increased bone resorption, but bone turnover has not previously been investigated in dogs with these conditions. The aim of this study was to determine whether a marker of bone resorption, ß-crosslaps, differs between dogs with CaOx urolithiasis and IH compared to controls. This retrospective, cross-sectional study used a canine specific ELISA to measure ß-crosslaps concentrations in stored frozen serum samples from 20 dogs with CaOx urolithiasis and IH and 20 breed-, sex-, and age-matched stone-free controls (18 Miniature Schnauzers, 14 Bichons Frise, and 8 Shih Tzus). Dogs with CaOx urolithiasis and IH had lower ß-crosslaps concentrations relative to controls (Pâ¯=â¯.0043), and ß-crosslaps had a moderate negative correlation with urinary calcium-to-creatinine ratios (râ¯=â¯-0.44, Pâ¯=â¯.0044). Miniature Schnauzers had lower ß-crosslaps concentrations than the other two breeds (Pâ¯=â¯.0035). The ELISA had acceptable intra-assay precision, but concentrations decreased when samples were repeatedly assayed over time. Assay recovery rates were also below acceptance criteria. In conclusion, Miniature Schnauzers, Bichons Frise, and Shih Tzus with CaOx urolithiasis and IH have evidence of decreased bone resorption compared to stone-free controls. This suggests that other causes of IH, such as intestinal hyperabsorption of calcium, underlie risk for CaOx urolithiasis in these breeds. Results should be confirmed in larger populations and with other ß-crosslaps assays and additional biomarkers of bone turnover. The stability of canine serum ß-crosslaps after freeze-thaw cycles and storage at various temperatures requires investigation.
Subject(s)
Bone Resorption/veterinary , Calcium Oxalate , Dog Diseases/etiology , Hypercalciuria/veterinary , Nephrolithiasis/veterinary , Animals , Bone Resorption/complications , Bone Resorption/pathology , Cross-Sectional Studies , Dog Diseases/pathology , Dogs , Female , Humans , Hypercalciuria/complications , Hypercalciuria/pathology , Male , Nephrolithiasis/complications , Nephrolithiasis/pathology , Retrospective StudiesABSTRACT
Familial hypomagnesemia with hypercalciuria and nephrocalcinosis is an autosomal-recessive renal tubular disorder characterized by excessive urinary losses of magnesium and calcium, bilateral nephrocalcinosis and progressive chronic renal failure in childhood or adolescence. The disease is caused by mutations in the tight-junction proteins claudin-16 and claudin-19 that are encoded by the CLDN16 and CLDN19 genes, respectively. Patients with CLDN19 mutations also are affected with severe ocular abnormalities. The aim of our study was to identify and characterize the molecular defects causing this disease in a Georgian girl and two Spanish siblings. Clinical and biochemical parameters were studied. The CLDN16 and CLDN19 genes were analyzed by DNA sequencing. The functional consequences of the identified mutations on pre-mRNA splicing were investigated using a minigene assay. Sequence analysis revealed that the patient from Georgia was homozygous for a novel mutation, c.602Gâ¯>â¯A; p.(G201E), in exon 4 of the CLDN16 gene. The two Spanish siblings were homozygous for a new CLDN19 mutation, c.388Gâ¯>â¯T; p.(G130C), located in exon 2, and both parents were heterozygous carriers of the mutation. Bioinformatics analysis predicted that the amino acid substitutions generated by these mutations were pathogenic. Functional studies showed that mutation c.388Gâ¯>â¯T also results in partial skipping of CLDN19 exon 2, which would imply significant alterations in the claudin-19 protein structure. Conversely, CLDN16 mutation c.602Gâ¯>â¯A had no effect on pre-mRNA splicing. Our study expands the genotypic classification of this rare disease and provides the first report of a CLDN19 mutation affecting splicing.
Subject(s)
Claudins/genetics , Hypercalciuria/genetics , Mutation , Nephrocalcinosis/genetics , Renal Tubular Transport, Inborn Errors/genetics , Adolescent , Alternative Splicing/genetics , Child, Preschool , DNA Mutational Analysis , Female , Humans , Hypercalciuria/pathology , Infant , Male , Nephrocalcinosis/pathology , Pedigree , Polymorphism, Single Nucleotide , Protein Isoforms/genetics , Renal Tubular Transport, Inborn Errors/pathology , SiblingsABSTRACT
The role of the calcium-sensing receptor (CaSR) as a regulator of parathyroid hormone secretion is well established, but its function in bone is less well defined. In an effort to elucidate the CaSR's skeletal role, bone tissue and material characteristics from patients with autosomal dominant hypocalcemia (ADH), a genetic form of primary hypoparathyroidism caused by CASR gain-of-function mutations, were compared to patients with postsurgical hypoparathyroidism (PSH). Bone structure and formation/resorption indices and mineralization density distribution (BMDD), were examined in transiliac biopsy samples from PSH (n = 13) and ADH (n = 6) patients by histomorphometry and quantitative backscatter electron imaging, respectively. Bone mineral density (BMD by DXA) and biochemical characteristics were measured at the time of the biopsy. Because both study groups comprised children and adults, all measured biopsy parameters and BMD outcomes were converted to Z-scores for comparison. Histomorphometric indices were normal and not different between ADH and PSH, with the exception of mineral apposition rate Z-score, which was higher in the ADH group. Similarly, average BMD Z-scores were normal and not different between ADH and PSH. Significant differences were observed for the BMDD: average Z-scores of mean and typical degree of mineralization (CaMean, CaPeak, respectively) were lower (p = 0.02 and p = 0.03, respectively), whereas the heterogeneity of mineralization (CaWidth) and percentage of lower mineralized areas (CaLow) were increased in ADH versus PSH (p = 0.01 and p = 0.002, respectively). The BMDD outcomes point toward a direct, PTH-independent role of the CaSR in the regulation of bone mineralization. © 2018 American Society for Bone and Mineral Research.
Subject(s)
Bone Density , Gain of Function Mutation , Hypercalciuria , Hypocalcemia , Hypoparathyroidism/congenital , Postoperative Complications , Receptors, Calcium-Sensing , Adolescent , Adult , Child , Female , Humans , Hypercalciuria/genetics , Hypercalciuria/metabolism , Hypercalciuria/pathology , Hypocalcemia/genetics , Hypocalcemia/metabolism , Hypocalcemia/pathology , Hypoparathyroidism/genetics , Hypoparathyroidism/metabolism , Hypoparathyroidism/pathology , Male , Middle Aged , Postoperative Complications/genetics , Postoperative Complications/metabolism , Postoperative Complications/pathology , Receptors, Calcium-Sensing/genetics , Receptors, Calcium-Sensing/metabolismABSTRACT
Research carried out during the past two-decades extended the understanding of actions of vitamin D, from regulating calcium and phosphate absorption and bone metabolism to many pleiotropic actions in organs and tissues in the body. Most observational and ecological studies report association of higher serum 25-hydroxyvitamin D [25(OH)D] concentrations with improved outcomes for several chronic, communicable and non-communicable diseases. Consequently, numerous agencies and scientific organizations have developed recommendations for vitamin D supplementation and guidance on optimal serum 25(OH)D concentrations. The bone-centric guidelines recommend a target 25(OH)D concentration of 20ng/mL (50nmol/L), and age-dependent daily vitamin D doses of 400-800IU. The guidelines focused on pleiotropic effects of vitamin D recommend a target 25(OH)D concentration of 30ng/mL (75nmol/L), and age-, body weight-, disease-status, and ethnicity dependent vitamin D doses ranging between 400 and 2000IU/day. The wise and balanced choice of the recommendations to follow depends on one's individual health outcome concerns, age, body weight, latitude of residence, dietary and cultural habits, making the regional or nationwide guidelines more applicable in clinical practice. While natural sources of vitamin D can raise 25(OH)D concentrations, relative to dietary preferences and latitude of residence, in the context of general population, these sources are regarded ineffective to maintain the year-round 25(OH)D concentrations in the range of 30-50ng/mL (75-125nmol/L). Vitamin D self-administration related adverse effects, such as hypercalcemia and hypercalciuria are rare, and usually result from taking extremely high doses of vitamin D for a prolonged time.
Subject(s)
Dietary Supplements , Vitamin D Deficiency/diet therapy , Vitamin D/analogs & derivatives , Vitamin D/administration & dosage , Adolescent , Adult , Age Factors , Body Weight , Feeding Behavior , Female , Humans , Hypercalcemia/blood , Hypercalcemia/chemically induced , Hypercalcemia/pathology , Hypercalciuria/blood , Hypercalciuria/chemically induced , Hypercalciuria/pathology , Infant , Infant, Newborn , Male , Middle Aged , Vitamin D/adverse effects , Vitamin D/blood , Vitamin D Deficiency/bloodABSTRACT
In humans, germline mutations in Trpm6 cause autosomal dominant hypomagnesemia with secondary hypocalcemia disorder. Loss of Trpm6 in mice also perturbs cellular magnesium homeostasis but additionally results in early embryonic lethality and neural tube closure defects. To define the mechanisms by which TRPM6 influences neural tube closure, we functionally characterized the role of TRPM6 during early embryogenesis in Xenopus laevis. The expression of Xenopus TRPM6 (XTRPM6) is elevated at the onset of gastrulation and is concentrated in the lateral mesoderm and ectoderm at the neurula stage. Loss of XTRPM6 produced gastrulation and neural tube closure defects. Unlike XTRPM6's close homologue XTRPM7, whose loss interferes with mediolateral intercalation, depletion of XTRPM6 but not XTRPM7 disrupted radial intercalation cell movements. A zinc-influx assay demonstrated that TRPM6 has the potential to constitute functional channels in the absence of TRPM7. The results of our study indicate that XTRPM6 regulates radial intercalation with little or no contribution from XTRPM7 in the region lateral to the neural plate, whereas XTRPM7 is mainly involved in regulating mediolateral intercalation in the medial region of the neural plate. We conclude that both TRPM6 and TRPM7 channels function cooperatively but have distinct and essential roles during neural tube closure.
Subject(s)
Embryonic Development/genetics , Neural Plate/growth & development , Neural Tube/growth & development , TRPM Cation Channels/genetics , Xenopus Proteins/genetics , Animals , Calcium/metabolism , Cell Movement/genetics , Gene Expression Regulation, Developmental , Germ-Line Mutation/genetics , Humans , Hypercalciuria/genetics , Hypercalciuria/metabolism , Hypercalciuria/pathology , Hypocalcemia/genetics , Hypocalcemia/metabolism , Hypocalcemia/pathology , Magnesium/metabolism , Nephrocalcinosis/genetics , Nephrocalcinosis/metabolism , Nephrocalcinosis/pathology , Neural Plate/metabolism , Renal Tubular Transport, Inborn Errors/genetics , Renal Tubular Transport, Inborn Errors/metabolism , Renal Tubular Transport, Inborn Errors/pathology , Xenopus laevisABSTRACT
Crystalluria is an imbalance between promoters and inhibitors; the first are the engine of the crystals and the second are made of substances. The biological assays make it possible to demonstrate one or more lithogenic factors observed during the lithiasis. The objective of our work was to study the impact of acidification or alkalization of the urine of lithiasic patients during a cristallurie assessment. All urine in the morning must be stored at room temperature and examined within two-three hours following voiding. 80 lithiasic patients (40 for urinary calcium and 40 for urinary uric acid) were selected from the results of cristalluria: more than 20 calcium-dependent crystals or uric acid crystal/mm3. For calcium and uric acid, concentration differences, after acidification and alkalization are observed: r2=0.43 (p<0.01) and r2=0.36 (p<0.01) respectively. In conclusion, in this lithiasic population with the presence of cystals, this pre-analytical stage is essential for urinary calcium and uric acid, making it possible to establish a diagnosis of certainly for the patient.
Subject(s)
Specimen Handling , Urinalysis/methods , Urine Specimen Collection/methods , Urolithiasis/urine , Calcium/analysis , Calcium/urine , Crystallization , Humans , Hydrogen-Ion Concentration , Hypercalciuria/pathology , Hypercalciuria/urine , Specimen Handling/methods , Specimen Handling/standards , Uric Acid/analysis , Uric Acid/urine , Urine Specimen Collection/standards , Urolithiasis/pathologyABSTRACT
INTRODUCTION: Hypercalciuria, hypocitraturia and cystinuria are the most common underlying metabolic stone abnormalities in children. The present study compared stone growth patterns, stone burden, and the risk of stone-related surgery among these underlying metabolic conditions. METHODS: A retrospective cohort of 356 children with renal stones, followed from 2000 to 2015, was studied. Differences among metabolic groups were determined using Kruskal-Wallis test; the Scheffé-test was used for multiple comparisons to determine differences among single groups. Independent sample t-test was used when adequate, given the sample size, and Chi-squared test was used for categorical variables. Stone growth rates were calculated as differences in diameter divided by time elapsed between U/Ss (mm/year). Logistic regression was performed to assess the effect of initial stone size on the likelihood of surgery. RESULTS: Median stone size at presentation was significantly different among groups, with cystinuria being the group with the largest proportion of stones >10 mm, while patients with stones <5 mm were likely to have a normal metabolic workup (P < 0.05). Stones with a higher growth rate were found in the operative group, while slower growing stones were mostly managed conservatively (3.4 mm/year vs 0.8 mm/year, respectively; P = 0.014). However, stone growth rates were not significantly different among metabolic groups. On the other hand, the rate of new stone formation in cystinuric patients at their first follow-up was 30.4%, which was significantly higher than in patients with hypercalciuria (16.3%) or with a normal metabolic workup (17.2%; P < 0.05). Compared with stones <5 mm, stones measuring 5-10 mm were more than four times more likely to result in surgery, whereas the likelihood of surgery for 10-20 mm or >20 mm stones was almost 16 or 34 times, respectively (P < 0.001). CONCLUSIONS: It is believed that this is the first study to evaluate stone growth patterns, stone burden and surgical risk among children with hypercalciuria, hypocitraturia and cystinuria. Cystinuric patients presented with larger stones at the time of diagnosis, higher new stone formation rates, and were at higher risk of surgery. While no significant difference of growth rate was found among metabolic groups, stones with a higher growth rate were significantly more likely to result in surgical treatment than slower growing stones. Initial stone size, location of largest stone, previous urinary tract infection, and patient's metabolic type significantly influenced the likelihood of a surgical intervention. Better understanding of the natural history ultimately helps surgeons and clinicians defining prognosis, treatment, and prevention plans for pediatric urolithiasis.
Subject(s)
Cystinuria/complications , Hypercalciuria/complications , Kidney Calculi/pathology , Kidney Calculi/surgery , Urolithiasis/pathology , Urolithiasis/surgery , Adolescent , Child , Child, Preschool , Cystinuria/pathology , Female , Humans , Hypercalciuria/pathology , Kidney Calculi/etiology , Male , Patient Selection , Retrospective Studies , Risk Factors , Urolithiasis/etiologyABSTRACT
The greatest risk factor for kidney stones is hypercalciuria, the etiology of which is largely unknown. A recent genome-wide association study (GWAS) linked hypercalciuria and kidney stones to a claudin-14 (CLDN14) risk haplotype. However, the underlying molecular mechanism was not delineated. Recently, renal CLDN14 expression was found to increase in response to increased plasma calcium, thereby inducing calciuria. We hypothesized therefore that some children with hypercalciuria and kidney stones harbor a CLDN14 variant that inappropriately increases gene expression. To test this hypothesis, we sequenced the CLDN14 risk haplotype in a cohort of children with idiopathic hypercalciuria and kidney stones. An intronic SNP was more frequent in affected children. Dual luciferase and cell-based assays demonstrated increased reporter or CLDN14 expression when this polymorphism was introduced. In silico studies predicted the SNP introduced a novel insulinoma-associated 1 (INSM1) transcription factor binding site. Consistent with this, repeating the dual luciferase assay in the presence of INSM1 further increased reporter expression. Our data suggest that children with the INSM1 binding site within the CLDN14 risk haplotype have a higher likelihood of hypercalciuria and kidney stones. Enhanced CLDN14 expression may play a role in the pathophysiology of their hypercalciuria.