Abstract
Background
Because of resistance to immunosuppressants in nephrotic syndrome and reduction of proteinuria relapses following renal transplantation, it seems that new horizons have arisen from mutational screening of the podocin gene. The aim of this study was to assess electronic microarray screening of the podocin mutation.
Methods
Twelve previously identified podocin mutations were screened by the electronic microarray method in known DNA samples and in patients (aged 5 months–18 years, n = 38) with steroid-resistant primary nephrotic syndrome, isolated proteinuria, end-stage renal disease secondary to idiopathic nephrotic syndrome, and proteinuria relapses following renal transplantation.
Results
DNA samples previously supplied to define the mutation profile for analysis and which were used as controls were completely and correctly detected by this method. None of the 12 mutations was detected in our patients. The duration of analysis for one mutation, including hybridization, was only 30 min for 38 cases.
Conclusion
Electronic microarray screening for NPHS2 mutations is not only rapid but also accurate. Previous identification of the mutation profile most often encountered in the investigated population is needed, however.
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References
Niaudet P (2004) Steroid-resistant idiopathic nephrotic syndrome in children. In: Avner ED, Harmon WE, Niaudet P (eds) Pediatric nephrology, 5th edn. Lippincott Williams & Wilkins, Philadelphia
Huber TB, Kottgen M, Schilling B et al (2001) Interaction with podocin facilitates nephrin signaling. J Biol Chem 276(45):41543–41546
Schwarz K, Simons M, Reiser J et al (2001) Podocin, a raft-associated component of the glomerular slit diaphragm interacts with CD2AP and nephrin. J Clin Invest 108(11):1621–1629
Kaplan JM, Kim SH, North KN et al (2000) Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis. Nat Genet 24(3):251–256
Caridi G, Perfumo F, Ghiggeri GM (2005) NPHS2 (Podocin) mutations in nephrotic syndrome. Clinical spectrum and fine mechanisms. Pediatr Res 57(5 Pt 2):54R–61R
Niaudet P (2004) Podocin and nephrotic syndrome: implications for the clinician. J Am Soc Nephrol 15(3):832–834
Westin L, Xu X, Miller C et al (2000) Anchored multiplex amplification on a microelectronic chip array. Nat Biotechnol 18(2):199–204
Gilles PN, Wu DJ, Foster CB et al (1999) Single nucleotide polymorphic discrimination by an electronic dot blot assay on semiconductor microchips. Nat Biotechnol 17(4):365–370
Mayo Clinic (1999) ASSAYS: beta testing of Nanogen’s NanoChip. Drug Discovery/Technology News, 12/1/1999
Corinne Antignac; Inserm U574 and Department of Genetics, Tour Lavoisier 6° etage, Necker Hospital 149 rue de Sevres, 75015 Paris, France
A report of the International Study of Kidney Disease in Children (1981) The primary nephrotic syndrome in children. Identification of patients with minimal change nephrotic syndrome from initial response to prednisone. J Pediatr 98(4):561–564
Boute N, Gribouval O, Roselli S et al (2000) NPHS2, encoding the glomerular protein podocin, is mutated in autosomal recessive steroid-resistant nephrotic syndrome. Nat Genet 24(4):349–354
Karle SM, Uetz B, Ronner V et al (2002) Novel mutations in NPHS2 detected in both familial and sporadic steroid-resistant nephrotic syndrome. J Am Soc Nephrol 13(2):388–393
Ruf RG, Lichtenberger A, Karle SM et al (2004) Patients with mutations in NPHS2 (podocin) do not respond to standard steroid treatment of nephrotic syndrome. J Am Soc Nephrol 15(3):722–732
Fuchshuber A, Jean G, Gribouval O et al (1995) Mapping a gene (SRN1) to chromosome 1q25–q31 in idiopathic nephrotic syndrome confirms a distinct entity of autosomal recessive nephrosis. Hum Mol Genet 4(11):2155–2158
Huber TB, Simons M, Hartleben B et al (2003) Molecular basis of the functional podocin–nephrin complex: mutations in the NPHS2 gene disrupt nephrin targeting to lipid raft microdomains. Hum Mol Genet 12(24):3397–3405
Berdeli A, Mir S, Yavascan O et al (2007) NPHS2 (podicin) mutations in Turkish children with idiopathic nephrotic syndrome. Pediatr Nephrol 22(12):2031–2040
Weber S, Gribouval O, Esquivel EL et al (2004) NPHS2 mutation analysis shows genetic heterogeneity of steroid resistant NS and low post-transplant recurrence. Kidney Int 66:571–579
Caridi G, Bertelli R, Di Duca M et al (2003) Broadening the spectrum of diseases related to podocin mutations. J Am Soc Nephrol 4:1278–1286
Hinkes B, Vlangos C, Heeringa S et al (2008) Specific podocin mutations correlate with age of onset in steroid-resistant nephrotic syndrome. J Am Soc Nephrol 19(2):365–371
Hinkes BG, Mucha B, Vlangos CN et al (2007) Nephrotic syndrome in the first year of life: two-thirds of cases are caused by mutations in 4 genes (NPHS1, NPHS2, WT1, and LAMB2. Pediatrics 119(4):e907–e919
Ozcakar ZB, Cengiz FB, Cakar N et al (2006) Analysis of NPHS2 mutations in Turkish steroid-resistant nephrotic syndrome patients. Pediatr Nephrol 21(8):1093–1096
Maruyama K, Iijima K, Ikeda M et al (2003) NPHS2 mutations in sporadic steroid-resistant nephrotic syndrome in Japanese children. Pediatr Nephrol 18(5):412–416
Acknowledgements
We express our gratitude to Ibrahim Aksu, Montwell Ltd, Ankara, for providing facilities, and continuous help and support, and to Corinnes Antignac, Inserm U574 and Department of Genetics, Necker Hospital, Paris, France, for providing the identified DNA samples.
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Sakallioglu, O., Gok, F., Kalman, S. et al. Electronic microarray screening of podocin mutations: a single-center study. Int Urol Nephrol 40, 1045–1051 (2008). https://doi.org/10.1007/s11255-008-9426-8
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DOI: https://doi.org/10.1007/s11255-008-9426-8