Cystine Nephrolithiasis Research Articles

#3320 GENETIC TESTING FOR MOLECULAR DIAGNOSIS OF NEPHROLITHIASIS AND NEPHROCALCINOSIS IN ADULT PATIENTS: A SINGLE-CENTER COHORT

Abstract Background and Aims Hereditary factors represent an important cause of nephrolithiasis which will generate stone disease in adult. Molecular analysis in patients with nephrolithiasis (NL) and/or nephrocalcinosis (NC) for a genetic mutation has become more accessible and the benefits are highlighted by an increasing number of publications. The aim of the paper was to study genetic screening for 36 NL and/or NC patients in order to identify the cases that can be confirmed by mutations in known kidney stone genes. Method Between 2020–2022 we included 35 adult patients with NL/NC with onset on pediatric age or in young adults, plus one of the following criteria: family history for NL/NC, indicative phenotype, recurrent NL. All the patients were diagnosed with NL and/or NC by ultrasound or computed tomography scan. Inform consent was signed and dated before blood samples and we performed genetic testing using nephrolithiasis panel (that include 45 genes) from 2 laboratories. In addition, we performed clinical assessment in a multidisciplinary team, underwent metabolic assessment, and caring out the genealogical tree. Results The study included 18 females and 17 males. The mean age of studied patients was 34.9 ± 10.3 years (range 18 – 54 years), although mean age of NL/NC diagnosis was 19.4 ± 12.0 years (range 0.5 – 34 years). 29 patients presented NL of any type, 12 patients presented both NL and NC, and 3 patients isolated NC. All the patients presented positive family history of NL/NC, 17 (48.5%) patients had pediatric age of onset, 17 (48.5%) patients presented indicative phenotype and 19 (54.2%) recurrent stone disease. Causative monogenic mutations were detected in 25 of 35 NL/NC. We identified 20 deleterious variants in 12 out of 45 analyzed genes. Genetic testing was positive with a definite diagnosis (had pathogenic variant) in 17 (48.6%) of cases, while 3 (8.6%) patients presented likely pathogenic variants, and 5 (14.3%) patients had variants of uncertain significance (VUS). In our cohort, the most common cause of kidney stone disease was cystine nephrolithiasis in 8 (22.8%) patients, followed by hereditary distal renal tubular acidosis in 4 patients, Dent disease in 3 patients, primary hyperoxaluria type 1 in 2 patients, familial hypomagnesaemia with hypercalciuria and nephrocalcinosis in 2 patients. Other causes of kidney stone disease included: renal hypouricemia type 1, hereditary hypophosphatemic rickets with hypercalciuria, primary mitochondrial disorders, autosomal dominant familial idiopathic hypercalciuria, Bartter syndrome type 3, and autosomal dominant tubulo-interstitial disease with hyperuricemia. Pathogenic mutations were detected in the following 5 dominant disease genes: SLC7A9 (4 patients), SLC4A1 (3 patients), ADCY10 (1 patient), HNF1B (2 patients), POLG (1 patient). Also, we identified pathogenic mutations in the following 7 recessive disease genes: CLCN5 (3 patients), AGXT (2 patients), CLDN16 (2 patients), SLC3A1 (3 patients), SLC34A3 (1 patient), WDR72 (1 patient), CLCNKB (1 patient). The mean eGFR for the study group was 71.2 ± 37.6 ml/min/1.72 m2. Seventeen patients presented eGFR < 60 ml/min/1.73 m2: seven patients CKD stage 3, four patients CKD stage 4, two patients CKD stage 5, and two patients were with renal replacement therapy. Conclusion Genetic kidney stone disease is an underdiagnosed condition. Although 48.5% of the patients had NL/NC onset on pediatric age, the molecular diagnosis was performed in adulthood, and, for some of them, when they suffer of advanced kidney disease. In our cohort, genetic testing had a high rate of positive molecular diagnosis of NL/NC due to selection criteria. Five of our patients presented VUS, but with disease-specific phenotype. We emphasize the importance of reporting these cases to generate additional evidence that could allow the reclassification of these variants. We conclude that the molecular diagnosis improves patient management, prevent or delay chronic kidney disease, offer possibility of genetic counseling and an extended screening to the family. Thus, our study showed the potential benefits of genetic testing, especially in high-risk groups for stone disease.

A Summary of Current Guidelines and Future Directions for Medical Management and Monitoring of Patients with Cystinuria

Cystinuria is the most common genetic cause of recurrent kidney stones. As the result of a genetic defect in proximal tubular reabsorption of filtered cystine, increased urine levels of the poorly soluble amino acid result in recurrent cystine nephrolithiasis. Recurrent cystine stones not only adversely affect the quality of patients suffering from cystinuria but also may result in chronic kidney disease (CKD) from recurrent renal injury. Thus, the mainstay of medical management revolves around prevention of stones. Recently published consensus statements on guidelines for managing cystinuria were released from both the United States and Europe. The purpose of this review is to summarize guidelines for medical management of patients with cystinuria, to provide new insight into the utility and clinical significance of cystine capacity-an assay for monitoring cystinuria, and to discuss future directions for research on treatment of cystinuria. We discuss future directions, including the potential use of cystine mimetics, gene therapy, V2-receptor blockers, and SGLT2 inhibitors, topics which have not appeared in more recent reviews. It is notable that in the absence of randomized, controlled trials, the recommendations cited here and in the guidelines are based on our best understanding of the disorder's pathophysiology, observational studies, and clinical experience.

Evaluation and Medical Management of Patients with Cystine Nephrolithiasis: A Consensus Statement.

Purpose: Cystinuria is a genetic disorder with both autosomal recessive and incompletely dominant inheritance. The disorder disrupts cystine and other dibasic amino acid transport in proximal tubules of the kidney, resulting in recurrent kidney stone formation. Currently, there are no consensus guidelines on evaluation and management of this disease. This article represents the consensus of the author panel and will provide clinicians with a stepwise framework for evaluation and clinical management of patients with cystinuria based on evidence in the existing literature. Materials and Methods: A search of MEDLINE®/PubMed® and Cochrane databases was performed using the following key words: "cystine nephrolithiasis," "cystinuria," "penicillamine, cystine," and "tiopronin, cystine." In total, as of May 2018, these searches yielded 2335 articles, which were then evaluated for their relevance to the topic of evaluation and management of cystinuria. Evidence was evaluated by the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) system. Results: Twenty-five articles on the topic of cystinuria or cystine nephrolithiasis were deemed suitable for inclusion in this study. The literature supports a logical evaluation process and step-wise treatment approach beginning with conservative measures: fluid intake and dietary modification. If stone formation recurs, proceed to pharmacotherapeutic options by first alkalinizing the urine and then using cystine-binding thiol drugs. Conclusions: The proposed clinical pathways provide a framework for efficient evaluation and treatment of patients with cystinuria, which should improve overall outcomes of this rare, but highly recurrent, form of nephrolithiasis.

Clinical Outcomes for Cystinuria Patients with Unilateral Versus Bilateral Cystine Stone Disease.

Cystinuria is a genetic disorder marked by elevated urinary cystine excretion and recurrent cystine nephrolithiasis. Interestingly, despite seemingly similar contralateral renal anatomy, a subset of cystinuric patients consistently form stones in only one kidney. The aim of this study is to evaluate clinical outcomes in unilateral vs bilateral cystine stone formers. We performed a retrospective case-control study of cystinuric patients evaluated and treated at the University of California, San Francisco between 1994 and 2015 and categorized patients as either unilateral or bilateral stone formers. Clinical presentation, baseline patient demographics, stone procedures, medical therapy regimens, and long-term renal function were compared between the two groups. A total of 42 cystine stone patients (22 female, 20 male) were included in the analysis. The median age at first presentation was 18.5 years and median age at study conclusion was 45.5 years. Two-thirds of patients (n = 28) had a history of bilateral stones, whereas one-third (n = 14) had unilateral stones. Medical therapy regimens were similar between groups. Despite an increased average number of lifetime surgeries (7.5 sessions for bilateral vs 3.7 sessions for unilateral, p < 0.05), there was no significant difference in medians of the most recent glomerular filtration rate when compared with unilateral stone formers (81.5 vs 95 mL/min, respectively; p = 0.28). The majority of cystinuric patients within our cohort form stones bilaterally during their lifetime, and require more surgical interventions than unilateral stone formers. Despite this, overall renal function is well preserved in unilateral and bilateral cystinuric stone formers treated with minimally invasive stone extraction procedures.

The Interaction of Thiol Drugs and Urine pH in the Treatment of Cystinuria

Pharmacological therapy for cystinuria consists of alkali salts to increase urine pH and thiol drugs to form soluble cysteine-drug complexes. The effect of alkalinizing urine on thiol drug activity has not been well studied. Urine samples were obtained from 5 healthy subjects and pH was adjusted (range 6.0 to 8.0) in each urine aliquot. Urine samples were incubated with cystine crystals at 37C for 5, 15 and 60 minutes, and 48 hours. We compared cystine solubility in urine samples spiked with thiol drugs at a final concentration of 2 mM to that in control urine samples at the various pH levels and time points. In samples incubated for 48 hours, which is the standard time frame for solubility studies, cystine solubility more than doubled with thiol drugs compared to control and did not depend on pH. However, when incubation time was shortened to 5 minutes, representing the dwell time of urine in the renal pelvis, the effect of thiol drugs to solubilize cystine greatly depended on urine pH with less cystine dissolved at lower pH. Increasing urine pH greatly increased the efficacy of thiol drugs to solubilize cystine in a clinically relevant time frame. These findings suggest that to maximize the benefit of thiol drugs alkali therapy should be used in conjunction with thiol drugs with the goal of keeping urine pH at 7.5 or above. Clinical trials are needed to confirm these in vitro findings.

Staghorn cystine stone in a 72-year-old recurrent calcium stone former

This case deals with the first diagnosis of Type B cystinuria with cystine nephrolithiasis in a 72-year-old male. Cystinuria is an inherited disease that consists of congenital abnormalities of renal and intestinal transport of dibasic amino acids. It often leads to frequent recurrent stone formation. Cystine stones most frequently occur in the 1st through 3rd decades of life with a decreased incidence in old age. This case shows that the first diagnosis of cystinuria may be made even in the 8th decade, without any family history, and in a patient with a history of recurrent calcium stone disease. Therefore, the chance of cystinuria must be always considered, even in older calcium stone formers.

UP-03.069 Urological Management of Cystine Nephrolithiasis; 20 Years of Experience in Four Different Centres

UP-03.069 Urological Management of Cystine Nephrolithiasis; 20 Years of Experience in Four Different Centres

Pharmacotherapy of urolithiasis: evidence from clinical trials

Urolithiasis is a worldwide problem with significant health and economic burdens. Medical therapy that alters the course of stone disease has enormous medical and financial impact. Urolithiasis is a final manifestation of a broad range of etiologies and pathogenesis. The modest progress in understanding the pathophysiology has hampered successful development of targeted therapy. Current regimens are based mostly on rational alteration of urinary biochemistry and physical chemistry to lower the risk of precipitation. In terms of pharmacotherapy, there are drugs to successfully improve hypercalciuria, hypocitraturia, aciduria, hyperuricosuria, and hypercystinuria. These agents have been proven to be effective in randomized controlled trials in improving urinary biochemical and physicochemical risk factors, as well as clinical outcomes. Although our current regimens have clearly improved the management and lives of stone formers, there are still clearly identifiable immense voids in the knowledge of pathophysiology of stone disease that can be filled with combined basic science and clinical studies.

Cystinuria: from diagnosis to follow-up

Cystinuria is an autosomal recessive disorder characterized by an impaired transport of cystine and dibasic aminoacids, lysine, arginine and ornithine in the proximal renal tubule and in the epithelial cells of the gastrointestinal tract. Recurrent cystine nephrolithiasis is the main clinical feature. Mutations in SLC3A1 and/or SLC7A9 genes, which are encoding respectively the rBAT and the b(0,+)AT proteins of the amino acid transport system, are responsible of this disorder thus inducing a high dibasic amino acid excretion. Diagnostic is based on stone analysis by infrared spectroscopy or microscopic examination of urine which may reveal typical cystine crystals. Quantitative cystine excretion, which may be assessed by aminoacid chromatography, is higher in cystinic patients. Molecular approach can identify mutations which are responsible of this pathology. Medical treatment is mainly based on hydratation and urine alkalinisation, with the addition of thiol derivative only in refractory cases. Follow-up based on pH and specific gravity determination in urine samples and cystine crystal volume measurement are used to optimally monitor the medical treatment of cystinuric patients. Even with medical management, long-term outcome is poor due to insufficient efficacy and low patient compliance. Many patients suffer from renal insufficiency as a result of recurrent stone formation and repeated surgical procedures.

Studies of urinary cystine precipitation in vitro: ontogeny of cystine nephrolithiasis and identification of meso-2,3-dimercaptosuccinic acid as a potential therapy for cystinuria

Children with fully recessive (Type I/I) cystinuria have a high risk of stone formation in the first decade of life. To assess the tendency for cystine to precipitate in individual urine samples, we developed an in vitro assay in which radiolabelled cystine (4 mM) was dissolved in urine at 37 °C after alkalization to pH 10. Samples were then brought to pH 5, cooled, and centrifuged. The % decrease in supernatant cpm was used as a measure of cystine precipitation (CP). CP varied widely among normal children (74% ± 34) whereas variability of repeated determinations on a single adult individual was modest (64% ± 3.3). The assay was used to compare various potential therapies for cystinuria. Precipitation of exogenous cystine from normal urine was strongly inhibited by addition of d-penicillamine (CP: 8% ± 3) or dimercaptosuccinic acid (DMSA) (CP: 5% ± 1), at urinary concentrations attained by standard oral doses of each drug. Mercaptopropionylglycine (MPG) was moderately effective (CP: 43% ± 9), whereas captopril was a weak inhibitor (CP: 63% ± 12). Precipitation of endogenous cystine (2191 μmol/L) from a cystinuric patient showed that DMSA and d-penicillamine were again highly effective compared to the other agents. In addition DMSA and penicillamine added to the same patient’s urine reduced the free cystine by 50% (as measured by automated amino acid analyzer) whereas MPG and captopril had no effect. In conclusion, DMSA is comparable to d-penicillamine as an in vitro inhibitor.

Laterality of symptomatic cystine calculi

ObjectivesTo evaluate patterns of laterality in cystine stone formation to determine whether phenotypic expression of this genetic disease is distributed equally in each kidney. Cystinuria is a genetic defect that may result in the formation of recurrent cystine calculi. Significant nongenetic factors may play a role in the manifestation of this disease. MethodsThirty-four patients seen in the Stone Center at the University of California, San Francisco were retrospectively evaluated for treatments and patterns of symptomatic stone formation between 1989 and 2002. A conservative surgical regimen and routine radiographic examinations were used. Treatments were used as a surrogate for development of symptomatic calculi. ResultsThe mean age at last follow-up was 38.2 years. The mean age at presentation was 18 years. The 34 patients underwent a total of 249 procedures, averaging 7.3 procedures per patient. Of the 34 patients, 29% required only unilateral surgery and formed unilateral symptomatic calculi exclusively (P <0.0001) during their lifetime. Of the 10 patients with unilateral stone formation, 2 (20%) were female and 8 (80%) were male. Of the 24 patients with bilateral stone formation, 12 (50%) were female and 12 (50%) were male. Of the 10 patients with unilateral stone formation, 6 developed left-sided calculi and 4 developed right-sided calculi. ConclusionsA subset of one third of patients with cystine stone formation will develop unilateral calculi exclusively during their lifetime. Understanding why the contralateral side did not form stones may provide insight into novel prophylactic regimens. The etiology of symptomatic unilateral cystine nephrolithiasis is unknown, but anatomic and external triggers should be considered.

Extracorporeal shock wave lithotripsy for cystine urolithiasis in children: outcome and complications.

The Siemens Lithostar Litotriptor was used to treat 6 children with cystine nephrolithiasis, previously treated by open surgery. Five children had renal calculi (3 multiple caliceal, 2 pelvis) and one had ureteral calculus. Stone size ranged from 0.2-2.5 cm in diameter, and stone burden was from 0.24 to 10.81 cm3 per kidney. From one to 4 ESWL sessions per unit were applied, with a total of 1,800 to 12,000 shock waves. The stone free rate at 3 months was 50%. A complete elimination was obtained with cystine stones in renal pelvis and ureter, however, up to 4 ESWL treatments failed in caliceal stones. Rather location of cystine calculi than previous surgery was associated with ESWL success rate. Two patients with positive urine cultures were successfully treated with appropriate antibiotics before ESWL was attempted. Perirenal hematoma was major complication demonstrated by radionuclide scintigraphy in one patient, and resolved spontaneously by 3 months. In the combined treatment of cystine urolithiasis in children ESWL, as auxillary procedure, was safe and effective in pelvis stone but failed in caliceal stones. Medical dissolution for retained fragments was found effective.

Ureteroscopic management of recurrent renal cystine calculi.

Patients with recurrent cystine nephrolithiasis oftentimes require multiple procedures for stone removal. As the majority of cystine stones are resistant to the effects of shockwave lithotripsy, repeat percutaneous surgery is often required and may cause renal damage. Moreover, repeat percutaneous access may become more difficult as perinephric fibrosis develops. Small-caliber ureteroscopes along with the holmium laser now enable routine intrarenal ureteroscopic access to symptomatic renal stones. Herein, we present our experience in managing recurrent renal cystine calculi using flexible ureterorenoscopy and assess whether such an approach may be used as an alternative to percutaneous surgery in selected patients. Three patients with large-volume (mean diameter 22 mm) renal cystine stones were managed with a 7.5F flexible ureterorenoscope combined with holmium laser lithotripsy to fragment the stones completely. The mean treatment time was 97 minutes, with successful fragmentation in all cases. Two of the three patients were completely stone free on follow-up intravenous urography, with the third patient having only small-volume residual fragments in a lower pole calix. All patients are currently asymptomatic and are being maintained on high oral fluid intake, urinary alkalization with potassium citrate, and alpha-mercaptopropionylglycine to reduce urinary cystine excretion. Flexible ureterorenoscopy with holmium laser lithotripsy provides a reasonable alternative for the management for recurrent cystine calculi in patients who are not candidates for repeat percutaneous procedures. Although it is time consuming, complete stone fragmentation, along with clearance of fragments, can be achieved in the majority of patients.

Genomic organization of a human cystine transporter gene (SLC3A1) and identification of novel mutations causing cystinuria

Cystinuria is a common inherited aminoaciduria that leads to recurrent cystine nephrolithiasis. Mutations in a gene encoding a renal amino acid transporter (SLC3A1) have been identified in patients with cystinuria establishing one molecular cause for the disease. To facilitate systematic screening of this gene for mutations, we have delineated the complete genomic organization of the SLC3A1 coding region using polymerase chain reaction strategies. The complete coding region of the gene is contained within a single yeast artificial chromosome clone and consists of 10 exons and 9 introns. Oligonucleotide primers capable of amplifying selected exons have been made and used in mutational analysis of DNA from 24 cystinuria probands. We illustrate the usefulness of this approach by identifying two novel SLC3A1 mutations. One novel mutation causes replacement of a highly conserved arginine residue (arginine-452) with tryptophan in the cytoplasmic loop between the putative third and fourth membrane spanning segments. A second previously unreported mutation results in replacement of a highly conserved tyrosine (tyrosine-461) residue with histidine in the same region of the protein. In addition, we detected three previously reported SLC3A1 mutations, R270X, 1500 +1/G to T, and M467T, the latter being present in approximately 20% of cystinuria chromosomes examined. Our findings provide a foundation for the development of more accessible diagnostic screening assays for detecting SLC3A1 mutations using patient genomic DNA, and also contribute to the emerging spectrum of cystinuria genotypes.

New technique for endourologic treatment of cystine nephrolithiasis

Cystine renal calculi are fragmented using a unique combination of ultrasonic and electrohydraulic lithotripsy. This endourologic application of a mining principle can be used to fragment any hard urinary stones.

Management of cystine nephrolithiasis by endourologic methods and shock-wave lithotripsy

Cystinuria is a rare cause of renal calculi, accounting for about 1 percent of cases. Herein we describe. our experience with a series of 10 patients with cystinuria and highlight the particular problems of the use of extracorporeal shock-wave lithotripsy (ESWL) to treat these types of stones and our resultant emphasis on percutaneous techniques.

The Spectrum of Metabolic Abnormalities in Patients with Cystine Nephrolithiasis

To elucidate the pathophysiology of mixed stone formation in cystinuria, 27 patients with documented cystine nephrolithiasis underwent an inpatient evaluation under a constant dietary regimen. All patients had homozygous cystinuria, since the daily urinary cystine excretion exceeded 250mg. per gm. creatinine. Hypercalciuria was noted in 5 patients (18.5 per cent), 4 of whom had fasting hypercalciuria. Hyperuricosuria was found in 6 patients (22.2 per cent) and it was not caused by a consumption of a diet rich in animal proteins, since urinary pH was higher and urinary sulfate lower than in control subjects. Serum uric acid was slightly lower and uric acid clearance was higher in hyperuricosuric patients than in control subjects. Hypocitraturia was found in 12 patients (44.4 per cent) and it was associated with defective renal acidification in 4 of 5 patients in whom it was tested.Thus, hypercalciuria, hyperuricosuria and hypocitraturia frequently accompany cystinuria in patients with cystine nephrolithiasis. These conditions might be renal in origin, rather than a result of dietary or environmental aberrations. They may contribute to the formation of calcium and uric acid stones, which sometimes complicate cystine nephrolithiasis.

CYSTINE NEPHROLITHIASIS

CYSTINE NEPHROLITHIASIS

"CYSTINE NEPHROLITHIASIS"

"CYSTINE NEPHROLITHIASIS"

"CYSTINE NEPHROLITHIASIS"

"CYSTINE NEPHROLITHIASIS"