Advertisement

Pediatric Nephrology

, Volume 33, Issue 10, pp 1741–1750 | Cite as

Use of genomic and functional analysis to characterize patients with steroid-resistant nephrotic syndrome

  • Thomas M. Kitzler
  • Nadezda Kachurina
  • Martin M. Bitzan
  • Elena Torban
  • Paul R. Goodyer
Original Article

Abstract

Background

Children with genetic causes of steroid-resistant nephrotic syndrome (SRNS) usually do well after renal transplantation, while some with idiopathic SRNS show recurrence due to a putative podocyte-toxic factor. Distinguishing different forms of SRNS based on clinical criteria has been difficult. The aim of our study was to test a novel approach that allows categorization of patients into clinically useful subgroups.

Methods

Seventeen patients with clinically confirmed SRNS were analyzed by next-generation sequencing (NGS) of 37 known SRNS genes and a functional assay of cultured human podocytes, which indirectly tests for toxicity of patients’ sera by evidenced loss of podocyte focal adhesion complex (FAC) number.

Results

We identified a pathogenic mutation in seven patients (41%). Sera from patients with monogenic SRNS caused mild loss of FAC number down to 73% compared to untreated controls, while sera from seven of the remaining ten patients with idiopathic SRNS caused significant FAC number loss to 43% (non-overlapping difference 30%, 95% CI 26–36%, P < 0.001). All patients with recurrent SRNS (n = 4) in the graft showed absence of podocyte gene mutations but significant FAC loss. Three patients had no mutation nor serum podocyte toxicity.

Conclusions

Our approach allowed categorization of patients into three subgroups: (1) patients with monogenic SRNS; (2) patients with idiopathic SRNS and marked serum podocyte toxicity; and (3) patients without identifiable genetic cause nor evidence of serum podocyte toxicity. Post-transplant SRNS recurrence risk appears to be low in groups 1 and 3, but high in group 2.

Keywords

Focal segmental glomerulosclerosis Steroid-resistant nephrotic syndrome Recurrent FSGS Next-generation sequencing Functional podocyte assay 

Notes

Acknowledgements

We thank all patients and their families at the Montreal Children’s Hospital and McMaster Children’s Hospital. We would also like to thank Dr. Vladimir Belostotsky who helped with recruitment and sample collection. Finally, we thank Dr. Moin Saleem for the gift of the human podocyte cell line.

Funding

This project received funds from the Pilot Project Competition of the Montreal Children’s Hospital Research Institute.

Compliance with ethical standards

This was a retrospective study in patients with established SRNS/FSGS and was approved by the local institutional review board. Informed consent was obtained from each participant or, where appropriate, the participant’s parents.

All procedures performed involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Conflict of interest

The authors declare that they have no conflicts of interest.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

467_2018_3995_MOESM1_ESM.docx (166 kb)
ESM 1 (DOCX 165 kb)

References

  1. 1.
    Eddy AA, Symons JM (2003) Nephrotic syndrome in childhood. Lancet 362:629–639CrossRefPubMedGoogle Scholar
  2. 2.
    Swaminathan S, Leung N, Lager DJ, Melton LJ 3rd, Bergstralh EJ, Rohlinger A, Fervenza FC (2006) Changing incidence of glomerular disease in Olmsted County, Minnesota: a 30-year renal biopsy study. Clin J Am Soc Nephrol 1:483–487CrossRefPubMedGoogle Scholar
  3. 3.
    Korbet SM, Schwartz MM, Lewis EJ (1994) Primary focal segmental glomerulosclerosis: clinical course and response to therapy. Am J Kidney Dis 23:773–783CrossRefPubMedGoogle Scholar
  4. 4.
    Newman WJ, Tisher CC, McCoy RC, Gunnells JC, Krueger RP, Clapp JR, Robinson RR (1976) Focal glomerular sclerosis: contrasting clinical patterns in children and adults. Medicine (Baltimore) 55:67–87CrossRefGoogle Scholar
  5. 5.
    White RH, Glasgow EF, Mills RJ (1970) Clinicopathological study of nephrotic syndrome in childhood. Lancet 1:1353–1359CrossRefPubMedGoogle Scholar
  6. 6.
    McCarthy HJ, Bierzynska A, Wherlock M, Ognjanovic M, Kerecuk L, Hegde S, Feather S, Gilbert RD, Krischock L, Jones C, Sinha MD, Webb NJ, Christian M, Williams MM, Marks S, Koziell A, Welsh GI, Saleem MA, Group RtUSS (2013) Simultaneous sequencing of 24 genes associated with steroid-resistant nephrotic syndrome. Clin J Am Soc Nephrol 8:637–648CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Sadowski CE, Lovric S, Ashraf S, Pabst WL, Gee HY, Kohl S, Engelmann S, Vega-Warner V, Fang H, Halbritter J, Somers MJ, Tan W, Shril S, Fessi I, Lifton RP, Bockenhauer D, El-Desoky S, Kari JA, Zenker M, Kemper MJ, Mueller D, Fathy HM, Soliman NA, Group SS, Hildebrandt F (2015) A single-gene cause in 29.5% of cases of steroid-resistant nephrotic syndrome. J Am Soc Nephrol 26:1279–1289CrossRefPubMedGoogle Scholar
  8. 8.
    Sen ES, Dean P, Yarram-Smith L, Bierzynska A, Woodward G, Buxton C, Dennis G, Welsh GI, Williams M, Saleem MA (2017) Clinical genetic testing using a custom-designed steroid-resistant nephrotic syndrome gene panel: analysis and recommendations. J Med Genet 54:795–804CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Abbott KC, Sawyers ES, Oliver JD 3rd, Ko CW, Kirk AD, Welch PG, Peters TG, Agodoa LY (2001) Graft loss due to recurrent focal segmental glomerulosclerosis in renal transplant recipients in the United States. Am J Kidney Dis 37:366–373CrossRefPubMedGoogle Scholar
  10. 10.
    Cheigh JS, Mouradian J, Susin M, Stubenbord WT, Tapia L, Riggio RR, Stenzel KH, Rubin AL (1980) Kidney transplant nephrotic syndrome: relationship between allograft histopathology and natural course. Kidney Int 18:358–365CrossRefPubMedGoogle Scholar
  11. 11.
    Choy BY, Chan TM, Lai KN (2006) Recurrent glomerulonephritis after kidney transplantation. Am J Transplant 6:2535–2542CrossRefPubMedGoogle Scholar
  12. 12.
    Kim EM, Striegel J, Kim Y, Matas AJ, Najarian JS, Mauer SM (1994) Recurrence of steroid-resistant nephrotic syndrome in kidney transplants is associated with increased acute renal failure and acute rejection. Kidney Int 45:1440–1445CrossRefPubMedGoogle Scholar
  13. 13.
    Tejani A, Stablein DH (1992) Recurrence of focal segmental glomerulosclerosis posttransplantation: a special report of the North American Pediatric Renal Transplant Cooperative Study. J Am Soc Nephrol 2:S258–S263PubMedGoogle Scholar
  14. 14.
    Gallon L, Leventhal J, Skaro A, Kanwar Y, Alvarado A (2012) Resolution of recurrent focal segmental glomerulosclerosis after retransplantation. N Engl J Med 366:1648–1649CrossRefPubMedGoogle Scholar
  15. 15.
    Lagrue G, Branellec A, Niaudet P, Heslan JM, Guillot F, Lang P (1991) Transmission of nephrotic syndrome to two neonates. Spontaneous regression. Presse Med 20:255–257PubMedGoogle Scholar
  16. 16.
    Torban E, Bitzan M, Goodyer P (2012) Recurrent focal segmental glomerulosclerosis: a discrete clinical entity. Int J Nephrol 2012:246128CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Saleem MA, O'Hare MJ, Reiser J, Coward RJ, Inward CD, Farren T, Xing CY, Ni L, Mathieson PW, Mundel P (2002) A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol 13:630–638PubMedGoogle Scholar
  18. 18.
    Kachurina N, Chung CF, Benderoff E, Babayeva S, Bitzan M, Goodyer P, Kitzler T, Matar D, Cybulsky AV, Alachkar N, Torban E (2016) Novel unbiased assay for circulating podocyte-toxic factors associated with recurrent focal segmental glomerulosclerosis. Am J Physiol Renal Physiol 310:F1148–F1156CrossRefPubMedGoogle Scholar
  19. 19.
    Babayeva S, Miller M, Zilber Y, El Kares R, Bernard C, Bitzan M, Goodyer P, Torban E (2011) Plasma from a case of recurrent idiopathic FSGS perturbs non-muscle myosin IIA (MYH9 protein) in human podocytes. Pediatr Nephrol 26:1071–1081CrossRefPubMedGoogle Scholar
  20. 20.
    Li H, Durbin R (2009) Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 25:1754–1760CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A, Garimella K, Altshuler D, Gabriel S, Daly M, DePristo MA (2010) The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res 20:1297–1303CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Bitzan M, Babayeva S, Vasudevan A, Goodyer P, Torban E (2012) TNFalpha pathway blockade ameliorates toxic effects of FSGS plasma on podocyte cytoskeleton and beta3 integrin activation. Pediatr Nephrol 27:2217–2226CrossRefPubMedGoogle Scholar
  23. 23.
    McKenzie LM, Hendrickson SL, Briggs WA, Dart RA, Korbet SM, Mokrzycki MH, Kimmel PL, Ahuja TS, Berns JS, Simon EE, Smith MC, Trachtman H, Michel DM, Schelling JR, Cho M, Zhou YC, Binns-Roemer E, Kirk GD, Kopp JB, Winkler CA (2007) NPHS2 variation in sporadic focal segmental glomerulosclerosis. J Am Soc Nephrol 18:2987–2995CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Boyer O, Benoit G, Gribouval O, Nevo F, Pawtowski A, Bilge I, Bircan Z, Deschenes G, Guay-Woodford LM, Hall M, Macher MA, Soulami K, Stefanidis CJ, Weiss R, Loirat C, Gubler MC, Antignac C (2010) Mutational analysis of the PLCE1 gene in steroid resistant nephrotic syndrome. J Med Genet 47:445–452CrossRefPubMedGoogle Scholar
  25. 25.
    Hinkes B, Wiggins RC, Gbadegesin R, Vlangos CN, Seelow D, Nurnberg G, Garg P, Verma R, Chaib H, Hoskins BE, Ashraf S, Becker C, Hennies HC, Goyal M, Wharram BL, Schachter AD, Mudumana S, Drummond I, Kerjaschki D, Waldherr R, Dietrich A, Ozaltin F, Bakkaloglu A, Cleper R, Basel-Vanagaite L, Pohl M, Griebel M, Tsygin AN, Soylu A, Muller D, Sorli CS, Bunney TD, Katan M, Liu J, Attanasio M, O'Toole JF, Hasselbacher K, Mucha B, Otto EA, Airik R, Kispert A, Kelley GG, Smrcka AV, Gudermann T, Holzman LB, Nurnberg P, Hildebrandt F (2006) Positional cloning uncovers mutations in PLCE1 responsible for a nephrotic syndrome variant that may be reversible. Nat Genet 38:1397–1405CrossRefPubMedGoogle Scholar
  26. 26.
    Little M, Wells C (1997) A clinical overview of WT1 gene mutations. Hum Mutat 9:209–225CrossRefPubMedGoogle Scholar
  27. 27.
    Tonna SJ, Needham A, Polu K, Uscinski A, Appel GB, Falk RJ, Katz A, Al-Waheeb S, Kaplan BS, Jerums G, Savige J, Harmon J, Zhang K, Curhan GC, Pollak MR (2008) NPHS2 variation in focal and segmental glomerulosclerosis. BMC Nephrol 9:13.  https://doi.org/10.1186/1471-2369-9-13 CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Chun MJ, Korbet SM, Schwartz MM, Lewis EJ (2004) Focal segmental glomerulosclerosis in nephrotic adults: presentation, prognosis, and response to therapy of the histologic variants. J Am Soc Nephrol 15:2169–2177CrossRefPubMedGoogle Scholar
  29. 29.
    Sethi S, Zand L, Nasr SH, Glassock RJ, Fervenza FC (2014) Focal and segmental glomerulosclerosis: clinical and kidney biopsy correlations. Clin Kidney J 7:531–537CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Thomas DB, Franceschini N, Hogan SL, Ten Holder S, Jennette CE, Falk RJ, Jennette JC (2006) Clinical and pathologic characteristics of focal segmental glomerulosclerosis pathologic variants. Kidney Int 69:920–926CrossRefPubMedGoogle Scholar
  31. 31.
    Mohey H, Thibaudin L, Laurent B, Berthoux F (2013) The podocin mutation R229Q and early recurrence (within the first year) of glomerular disease after renal transplantation. Ann Transplant 18:436–442CrossRefPubMedGoogle Scholar
  32. 32.
    Weber S, Tonshoff B (2005) Recurrence of focal-segmental glomerulosclerosis in children after renal transplantation: clinical and genetic aspects. Transplantation 80:S128–S134CrossRefPubMedGoogle Scholar
  33. 33.
    Jungraithmayr TC, Hofer K, Cochat P, Chernin G, Cortina G, Fargue S, Grimm P, Knueppel T, Kowarsch A, Neuhaus T, Pagel P, Pfeiffer KP, Schafer F, Schonermarck U, Seeman T, Toenshoff B, Weber S, Winn MP, Zschocke J, Zimmerhackl LB (2011) Screening for NPHS2 mutations may help predict FSGS recurrence after transplantation. J Am Soc Nephrol 22:579–585CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Bierzynska A, McCarthy HJ, Soderquest K, Sen ES, Colby E, Ding WY, Nabhan MM, Kerecuk L, Hegde S, Hughes D, Marks S, Feather S, Jones C, Webb NJ, Ognjanovic M, Christian M, Gilbert RD, Sinha MD, Lord GM, Simpson M, Koziell AB, Welsh GI, Saleem MA (2017) Genomic and clinical profiling of a national nephrotic syndrome cohort advocates a precision medicine approach to disease management. Kidney Int 91:937–947CrossRefPubMedGoogle Scholar
  35. 35.
    Konigshausen E, Sellin L (2016) Circulating permeability factors in primary focal segmental glomerulosclerosis: a review of proposed candidates. Biomed Res Int 2016:3765608.  https://doi.org/10.1155/2016/3765608 CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Kronbichler A, Saleem MA, Meijers B, Shin JI (2016) Soluble urokinase receptors in focal segmental glomerulosclerosis: a review on the scientific point of view. J Immunol Res 2016:2068691CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Pedigo CE, Ducasa GM, Leclercq F, Sloan A, Mitrofanova A, Hashmi T, Molina-David J, Ge M, Lassenius MI, Forsblom C, Lehto M, Groop PH, Kretzler M, Eddy S, Martini S, Reich H, Wahl P, Ghiggeri G, Faul C, Burke GW 3rd, Kretz O, Huber TB, Mendez AJ, Merscher S, Fornoni A (2016) Local TNF causes NFATc1-dependent cholesterol-mediated podocyte injury. J Clin Invest 126:3336–3350CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Savin VJ, Sharma M, Zhou J, Genochi D, Sharma R, Srivastava T, Ilahe A, Budhiraja P, Gupta A, McCarthy ET (2017) Multiple targets for novel therapy of FSGS associated with circulating permeability factor. Biomed Res Int 2017:6232616CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Wei C, El Hindi S, Li J, Fornoni A, Goes N, Sageshima J, Maiguel D, Karumanchi SA, Yap HK, Saleem M, Zhang Q, Nikolic B, Chaudhuri A, Daftarian P, Salido E, Torres A, Salifu M, Sarwal MM, Schaefer F, Morath C, Schwenger V, Zeier M, Gupta V, Roth D, Rastaldi MP, Burke G, Ruiz P, Reiser J (2011) Circulating urokinase receptor as a cause of focal segmental glomerulosclerosis. Nat Med 17:952–960CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Taneda S, Honda K, Ohno M, Uchida K, Nitta K, Oda H (2015) Podocyte and endothelial injury in focal segmental glomerulosclerosis: an ultrastructural analysis. Virchows Arch 467:449–458CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© IPNA 2018

Authors and Affiliations

  • Thomas M. Kitzler
    • 1
  • Nadezda Kachurina
    • 2
  • Martin M. Bitzan
    • 3
  • Elena Torban
    • 2
  • Paul R. Goodyer
    • 3
  1. 1.Department of Medical Genetics, The Montreal Children’s HospitalMcGill University Health CenterMontrealCanada
  2. 2.Department of MedicineMcGill University and McGill University Health CenterMontrealCanada
  3. 3.Division of Nephrology, The Montreal Children’s HospitalMcGill University Health CenterMontrealCanada

Personalised recommendations