Skip to main content
SpringerLink
Log in

Identification of 47 novel mutations in patients with Alport syndrome and thin basement membrane nephropathy

  • Original Article
  • Published:
Pediatric Nephrology Aims and scope Submit manuscript

Abstract

Background

Alport syndrome (ATS) is a progressive hereditary nephropathy characterized by hematuria and proteinuria. It can be associated with extrarenal manifestations. In contrast, thin basement membrane nephropathy (TBMN) is characterized by microscopic hematuria, is largely asymptomatic, and is rarely associated with proteinuria and end-stage renal disease. Mutations have been identified in the COL4A5 gene in ATS and in the COL4A3 and COL4A4 genes in ATS and TBMN. To date, more than 1000 different mutations in COL4A5, COL4A3, and COL4A4 are known.

Methods

In this study mutational analysis by exon sequencing and multiplex ligation-dependent probe amplification was performed in a large European cohort of families with ATS and TBMN.

Results

Molecular diagnostic testing of 216 individuals led to the detection of 47 novel mutations, thereby expanding the spectrum of known mutations causing ATS and TBMN by up to 10 and 6 %, respectively, depending on the database. Remarkably, a high number of ATS patients with only single mutations in COL4A3 and COL4A4 were identified. Additionally, three ATS patients presented with synonymous sequence variants that possible affect correct mRNA splicing, as suggested by in silico analysis.

Conclusions

The results of this study clearly broaden the genotypic spectrum of known mutations for ATS and TBMN, which will in turn now facilitate future studies into genotype–phenotype correlations. Further studies should also examine the significance of single heterozygous mutations in COL4A3 and COL4A4 and of synonymous sequence variants associated with ATS.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Noone D, Licht C (2013) An update on the pathomechanisms and future therapies of Alport syndrome. Pediatr Nephrol 28:1025–1036

    Article  PubMed  Google Scholar 

  2. Marcocci E, Uliana V, Bruttini M, Artuso R, Silengo MC, Zerial M, Bergesio F, Amoroso A, Savoldi S, Pennesi M, Giachino D, Rombola G, Fogazzi GB, Rosatelli C, Martinhago CD, Carmellini M, Mancini R, Di Costanzo G, Longo I, Renieri A, Mari F (2009) Autosomal dominant Alport syndrome: molecular analysis of the COL4A4 gene and clinical outcome. Nephrol Dial Transplant 24:1464–1471

    Article  CAS  PubMed  Google Scholar 

  3. Nagel M, Nagorka S, Gross O (2005) Novel COL4A5, COL4A4, and COL4A3 mutations in Alport syndrome. Hum Mutat 26:60

    Article  PubMed  Google Scholar 

  4. Fallerini C, Dosa L, Tita R, Del Prete D, Feriozzi S, Gai G, Clementi M, La Manna A, Miglietti N, Mancini R, Mandrile G, Ghiggeri GM, Piaggio G, Brancati F, Diano L, Frate E, Pinciaroli AR, Giani M, Castorina P, Bresin E, Giachino D, De Marchi M, Mari F, Bruttini M, Renieri A, Ariani F (2014) Unbiased next generation sequencing analysis confirms the existence of autosomal dominant Alport syndrome in a relevant fraction of cases. Clin Genet 86:252–257

    Article  CAS  PubMed  Google Scholar 

  5. Hou P, Chen Y, Ding J, Li G, Zhang H (2007) A novel mutation of COL4A3 presents a different contribution to Alport syndrome and thin basement membrane nephropathy. Am J Nephrol 27:538–544

    Article  CAS  PubMed  Google Scholar 

  6. Cook C, Friedrich CA, Baliga R (2008) Novel COL4A3 mutations in African American siblings with autosomal recessive Alport syndrome. Am J Kidney Dis 51:e25–e28

    Article  PubMed  Google Scholar 

  7. Longo I, Scala E, Mari F, Caselli R, Pescucci C, Mencarelli MA, Speciale C, Giani M, Bresin E, Caringella DA, Borochowitz ZU, Siriwardena K, Winship I, Renieri A, Meloni I (2006) Autosomal recessive Alport syndrome: an in-depth clinical and molecular analysis of five families. Nephrol Dial Transplant 21:665–671

    Article  CAS  PubMed  Google Scholar 

  8. Baek JI, Choi SJ, Park SH, Choi JY, Kim CD, Kim YL, Kim UK (2009) Identification of novel variants in the COL4A4 gene in Korean patients with thin basement membrane nephropathy. Indian J Med Res 129:525–533

    CAS  PubMed  Google Scholar 

  9. Fujinaga S, Kaneko K, Ohtomo Y, Murakami H, Takemoto M, Takada M, Shimizu T, Yamashiro Y (2006) Thin basement membrane nephropathy associated with minimal change disease in a 15-year-old boy. Pediatr Nephrol 21:277–280

    Article  PubMed  Google Scholar 

  10. Voskarides K, Pierides A, Deltas C (2008) COL4A3/COL4A4 mutations link familial hematuria and focal segmental glomerulosclerosis. glomerular epithelium destruction via basement membrane thinning? Connect Tissue Res 49:283–288

    Article  CAS  PubMed  Google Scholar 

  11. Tryggvason K, Patrakka J (2006) Thin basement membrane nephropathy. J Am Soc Nephrol 17:813–822

    Article  CAS  PubMed  Google Scholar 

  12. Cangiotti AM, Sessa A, Meroni M, Montironi R, Ragaiolo M, Mambelli V, Cinti S (1996) Evolution of glomerular basement membrane lesions in a male patient with Alport syndrome: ultrastructural and morphometric study. Nephrol Dial Transplant 11:1829–1834

    Article  CAS  PubMed  Google Scholar 

  13. Savige J, Rana K, Tonna S, Buzza M, Dagher H, Wang YY (2003) Thin basement membrane nephropathy. Kidney Int 64:1169–1178

    Article  PubMed  Google Scholar 

  14. Gubler MC (2007) Diagnosis of Alport syndrome without biopsy? Pediatr Nephrol 22:621–625

    Article  PubMed  Google Scholar 

  15. Savige J, Gregory M, Gross O, Kashtan C, Ding J, Flinter F (2013) Expert guidelines for the management of Alport syndrome and thin basement membrane nephropathy. J Am Soc Nephrol 24:364–375

    Article  CAS  PubMed  Google Scholar 

  16. Mencarelli MA, Heidet L, Storey H, van Geel M, Knebelmann B, Fallerini C, Miglietti N, Antonucci MF, Cetta F, Sayer JA, van den Wijngaard A, Yau S, Mari F, Bruttini M, Ariani F, Dahan K, Smeets B, Antignac C, Flinter F, Renieri A (2015) Evidence of digenic inheritance in Alport syndrome. J Med Genet 52:163–174

    Article  CAS  PubMed  Google Scholar 

  17. Bekheirnia MR, Reed B, Gregory MC, McFann K, Shamshirsaz AA, Masoumi A, Schrier RW (2010) Genotype-phenotype correlation in X-linked Alport syndrome. J Am Soc Nephrol 21:876–883

  18. Storey H, Savige J, Sivakumar V, Abbs S, Flinter FA (2013) COL4A3/COL4A4 mutations and features in individuals with autosomal recessive Alport syndrome. J Am Soc Nephrol 24:1945–1954

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Jais JP, Knebelmann B, Giatras I, De Marchi M, Rizzoni G, Renieri A, Weber M, Gross O, Netzer KO, Flinter F, Pirson Y, Verellen C, Wieslander J, Persson U, Tryggvason K, Martin P, Hertz JM, Schroder C, Sanak M, Krejcova S, Carvalho MF, Saus J, Antignac C, Smeets H, Gubler MC (2000) X-linked Alport syndrome: natural history in 195 families and genotype-phenotype correlations in males. J Am Soc Nephrol 11:649–657

  20. Gross O, Netzer KO, Lambrecht R, Seibold S, Weber M (2002) Meta-analysis of genotype-phenotype correlation in X-linked Alport syndrome: impact on clinical counselling. Nephrol Dial Transplant 17:1218–1227

    Article  PubMed  Google Scholar 

  21. Jais JP, Knebelmann B, Giatras I, De Marchi M, Rizzoni G, Renieri A, Weber M, Gross O, Netzer KO, Flinter F, Pirson Y, Dahan K, Wieslander J, Persson U, Tryggvason K, Martin P, Hertz JM, Schroder C, Sanak M, Carvalho MF, Saus J, Antignac C, Smeets H, Gubler MC (2003) X-linked Alport syndrome: natural history and genotype-phenotype correlations in girls and women belonging to 195 families: a “European Community Alport Syndrome Concerted Action” study. J Am Soc Nephrol 14:2603–2610

    Article  PubMed  Google Scholar 

  22. Study Group Members of the Gesellschaft fur Padiatrische N, Gross O, Licht C, Anders HJ, Hoppe B, Beck B, Tonshoff B, Hocker B, Wygoda S, Ehrich JH, Pape L, Konrad M, Rascher W, Dotsch J, Muller-Wiefel DE, Hoyer P, Knebelmann B, Pirson Y, Grunfeld JP, Niaudet P, Cochat P, Heidet L, Lebbah S, Torra R, Friede T, Lange K, Muller GA, Weber M (2012) Early angiotensin-converting enzyme inhibition in Alport syndrome delays renal failure and improves life expectancy. Kidney Int 81:494–501

    Article  Google Scholar 

  23. Proesmans W, Van Dyck M (2004) Enalapril in children with Alport syndrome. Pediatr Nephrol 19:271–275

    Article  PubMed  Google Scholar 

  24. Rana K, Tonna S, Wang YY, Sin L, Lin T, Shaw E, Mookerjee I, Savige J (2007) Nine novel COL4A3 and COL4A4 mutations and polymorphisms identified in inherited membrane diseases. Pediatr Nephrol 22:652–657

    Article  PubMed  Google Scholar 

  25. Pierides A, Voskarides K, Athanasiou Y, Ioannou K, Damianou L, Arsali M, Zavros M, Pierides M, Vargemezis V, Patsias C, Zouvani I, Elia A, Kyriacou K, Deltas C (2009) Clinico-pathological correlations in 127 patients in 11 large pedigrees, segregating one of three heterozygous mutations in the COL4A3/COL4A4 genes associated with familial haematuria and significant late progression to proteinuria and chronic kidney disease from focal segmental glomerulosclerosis. Nephrol Dial Transplant 24:2721–2729

    Article  CAS  PubMed  Google Scholar 

  26. Voskarides K, Damianou L, Neocleous V, Zouvani I, Christodoulidou S, Hadjiconstantinou V, Ioannou K, Athanasiou Y, Patsias C, Alexopoulos E, Pierides A, Kyriacou K, Deltas C (2007) COL4A3/COL4A4 mutations producing focal segmental glomerulosclerosis and renal failure in thin basement membrane nephropathy. J Am Soc Nephrol 18:3004–3016

    Article  CAS  PubMed  Google Scholar 

  27. Hertz JM, Thomassen M, Storey H, Flinter F (2015) Clinical utility gene card for: Alport syndrome - update 2014. Eur J Hum Genet. doi:10.1038/ejhg.2014.254

  28. Wang YY, Rana K, Tonna S, Lin T, Sin L, Savige J (2004) COL4A3 mutations and their clinical consequences in thin basement membrane nephropathy (TBMN). Kidney Int 65:786–790

    Article  CAS  PubMed  Google Scholar 

  29. Heidet L, Arrondel C, Forestier L, Cohen-Solal L, Mollet G, Gutierrez B, Stavrou C, Gubler MC, Antignac C (2001) Structure of the human type IV collagen gene COL4A3 and mutations in autosomal Alport syndrome. J Am Soc Nephrol 12:97–106

    CAS  PubMed  Google Scholar 

  30. Chatterjee R, Hoffman M, Cliften P, Seshan S, Liapis H, Jain S (2013) Targeted exome sequencing integrated with clinicopathological information reveals novel and rare mutations in atypical, suspected and unknown cases of Alport syndrome or proteinuria. PLoS One 8:e76360

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Lemmink HH, Nillesen WN, Mochizuki T, Schroder CH, Brunner HG, van Oost BA, Monnens LA, Smeets HJ (1996) Benign familial hematuria due to mutation of the type IV collagen alpha4 gene. J Clin Invest 98:1114–1118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Knebelmann B, Deschenes G, Gros F, Hors MC, Grunfeld JP, Zhou J, Tryggvason K, Gubler MC, Antignac C (1992) Substitution of arginine for glycine 325 in the collagen alpha 5 (IV) chain associated with X-linked Alport syndrome: characterization of the mutation by direct sequencing of PCR-amplified lymphoblast cDNA fragments. Am J Hum Genet 51:135–142

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Martin P, Heiskari N, Zhou J, Leinonen A, Tumelius T, Hertz JM, Barker D, Gregory M, Atkin C, Styrkarsdottir U, Neumann H, Springate J, Shows T, Pettersson E, Tryggvason K (1998) High mutation detection rate in the COL4A5 collagen gene in suspected Alport syndrome using PCR and direct DNA sequencing. J Am Soc Nephrol 9:2291–2301

    CAS  PubMed  Google Scholar 

  34. Beicht S, Strobl-Wildemann G, Rath S, Wachter O, Alberer M, Kaminsky E, Weber LT, Hinrichsen T, Klein HG, Hoefele J (2013) Next generation sequencing as a useful tool in the diagnostics of mosaicism in Alport syndrome. Gene 526:474–477

    Article  CAS  PubMed  Google Scholar 

  35. Boye E, Flinter F, Zhou J, Tryggvason K, Bobrow M, Harris A (1995) Detection of 12 novel mutations in the collagenous domain of the COL4A5 gene in Alport syndrome patients. Hum Mutat 5:197–204

    Article  CAS  PubMed  Google Scholar 

  36. Plant KE, Green PM, Vetrie D, Flinter FA (1999) Detection of mutations in COL4A5 in patients with Alport syndrome. Hum Mutat 13:124–132

    Article  CAS  PubMed  Google Scholar 

  37. Massella L, Gangemi C, Giannakakis K, Crisafi A, Faraggiana T, Fallerini C, Renieri A, Muda AO, Emma F (2013) Prognostic value of glomerular collagen IV immunofluorescence studies in male patients with X-linked Alport syndrome. Clin J Am Soc Nephrol 8:749–755

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Hashimura Y, Nozu K, Kaito H, Nakanishi K, Fu XJ, Ohtsubo H, Hashimoto F, Oka M, Ninchoji T, Ishimori S, Morisada N, Matsunoshita N, Kamiyoshi N, Yoshikawa N, Iijima K (2014) Milder clinical aspects of X-linked Alport syndrome in men positive for the collagen IV alpha5 chain. Kidney Int 85:1208–1213

    Article  CAS  PubMed  Google Scholar 

  39. Wang F, Zhao D, Ding J, Zhang H, Zhang Y, Yu L, Xiao H, Yao Y, Zhong X, Wang S (2012) Skin biopsy is a practical approach for the clinical diagnosis and molecular genetic analysis of X-linked Alport’s syndrome. J Mol Diagn 14:586–593

    Article  CAS  PubMed  Google Scholar 

  40. Hertz JM, Juncker I, Marcussen N (2008) MLPA and cDNA analysis improves COL4A5 mutation detection in X-linked Alport syndrome. Clin Genet 74:522–530

    Article  CAS  PubMed  Google Scholar 

  41. Knebelmann B, Breillat C, Forestier L, Arrondel C, Jacassier D, Giatras I, Drouot L, Deschenes G, Grunfeld JP, Broyer M, Gubler MC, Antignac C (1996) Spectrum of mutations in the COL4A5 collagen gene in X-linked Alport syndrome. Am J Hum Genet 59:1221–1232

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Gross O, Friede T, Hilgers R, Gorlitz A, Gavenis K, Ahmed R, Durr U (2012) Safety and efficacy of the ACE-inhibitor Ramipril in Alport syndrome: the double-blind, randomized, placebo-controlled, multicenter phase III EARLY PRO-TECT Alport trial in pediatric patients. ISRN Pediatr 2012:436046

    Article  PubMed  PubMed Central  Google Scholar 

  43. Miner JH, Baigent C, Flinter F, Gross O, Judge P, Kashtan CE, Lagas S, Savige J, Blatt D, Ding J, Gale DP, Midgley JP, Povey S, Prunotto M, Renault D, Skelding J, Turner AN, Gear S (2014) The 2014 International Workshop on Alport Syndrome. Kidney Int 86:679–684

    Article  PubMed  PubMed Central  Google Scholar 

  44. King K, Flinter FA, Nihalani V, Green PM (2002) Unusual deep intronic mutations in the COL4A5 gene cause X linked Alport syndrome. Hum Genet 111:548–554

    Article  CAS  PubMed  Google Scholar 

  45. Tryggvason K, Zhou J, Hostikka SL, Shows TB (1993) Molecular genetics of Alport syndrome. Kidney Int 43:38–44

    Article  CAS  PubMed  Google Scholar 

  46. Anton-Martin P, Aparicio Lopez C, Ramiro-Leon S, Santillan Garzon S, Santos-Simarro F, Gil-Fournier B (2012) Alport syndrome: de novo mutation in the COL4A5 gene converting glycine 1205 to valine. Clin Med Insights Pediatr 6:41–49

    Article  PubMed  PubMed Central  Google Scholar 

  47. Strachan T, Read AP (1999) Human molecular genetics. Wiley-Liss, New York

  48. Kashtan CE (1999) Alport syndrome. An inherited disorder of renal, ocular, and cochlear basement membranes. Medicine (Baltimore) 78:338–360

    Article  CAS  Google Scholar 

  49. Slajpah M, Gorinsek B, Berginc G, Vizjak A, Ferluga D, Hvala A, Meglic A, Jaksa I, Furlan P, Gregoric A, Kaplan-Pavlovcic S, Ravnik-Glavac M, Glavac D (2007) Sixteen novel mutations identified in COL4A3, COL4A4, and COL4A5 genes in Slovenian families with Alport syndrome and benign familial hematuria. Kidney Int 71:1287–1295

    Article  CAS  PubMed  Google Scholar 

  50. Demosthenous P, Voskarides K, Stylianou K, Hadjigavriel M, Arsali M, Patsias C, Georgaki E, Zirogiannis P, Stavrou C, Daphnis E, Pierides A, Deltas C, Hellenic Nephrogenetics Research C (2012) X-linked Alport syndrome in Hellenic families: phenotypic heterogeneity and mutations near interruptions of the collagen domain in COL4A5. Clin Genet 81:240–248

    Article  CAS  PubMed  Google Scholar 

  51. Deltas C, Pierides A, Voskarides K (2012) The role of molecular genetics in diagnosing familial hematuria(s). Pediatr Nephrol 27:1221–1231

    Article  PubMed  PubMed Central  Google Scholar 

  52. Pierides A, Voskarides K, Kkolou M, Hadjigavriel M, Deltas C (2013) X-linked, COL4A5 hypomorphic Alport mutations such as G624D and P628L may only exhibit thin basement membrane nephropathy with microhematuria and late onset kidney failure. Hippokratia 17:207–213

    CAS  PubMed  PubMed Central  Google Scholar 

  53. Barker DF, Denison JC, Atkin CL, Gregory MC (2001) Efficient detection of Alport syndrome COL4A5 mutations with multiplex genomic PCR-SSCP. Am J Med Genet 98:148--160

Download references

Acknowledgments

We sincerely thank the patients and the physicians for participation in this study.

Author information

Author notes

    Authors and Affiliations

    1. Pediatric Nephrology, Pediatrics II, University of Duisburg-Essen, Essen, Germany

      Stefanie Weber, Katja Strasser & Peter F. Hoyer

    2. Center for Human Genetics and Laboratory Diagnostics Dr. Klein, Dr. Rost and Colleagues, Martinsried, Germany

      Sabine Rath, Achim Kittke, Sonja Beicht, Hanns-Georg Klein & Julia Hoefele

    3. Department of Infectious Diseases and Tropical Medicine, Ludwig-Maximilians University, Munich, Germany

      Martin Alberer

    4. Pediatric Nephrology, University Children’s Hospital, Ludwig-Maximilians University, Munich, Germany

      Bärbel Lange-Sperandio & Sabine Ponsel

    5. Pediatric Nephrology, University Children’s Hospital, Cologne, Germany

      Marcus R. Benz & Lutz T. Weber

    6. Institute of Human Genetics, Technische Universität München, Munich, Germany

      Julia Hoefele

    Authors
    1. Stefanie Weber
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Katja Strasser
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Sabine Rath
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Achim Kittke
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Sonja Beicht
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Martin Alberer
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Bärbel Lange-Sperandio
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Peter F. Hoyer
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Marcus R. Benz
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Sabine Ponsel
      View author publications

      You can also search for this author in PubMed Google Scholar

    11. Lutz T. Weber
      View author publications

      You can also search for this author in PubMed Google Scholar

    12. Hanns-Georg Klein
      View author publications

      You can also search for this author in PubMed Google Scholar

    13. Julia Hoefele
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Julia Hoefele.

    Ethics declarations

    Approval for this study was obtained from the Ethics Committee of the Ludwig-Maximilians University, Munich, Germany. Written informed consent was obtained.

    Conflict of interest

    The authors declare no conflict of interest.

    Additional information

    Stefanie Weber and Katja Strasser contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Weber, S., Strasser, K., Rath, S. et al. Identification of 47 novel mutations in patients with Alport syndrome and thin basement membrane nephropathy. Pediatr Nephrol 31, 941–955 (2016). https://doi.org/10.1007/s00467-015-3302-4

    Download citation

    • Received:

    • Revised:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00467-015-3302-4

    Keywords

    Search

    Navigation