Advertisement

The Indian Journal of Pediatrics

, Volume 86, Issue 8, pp 692–699 | Cite as

Glucose Phosphate Isomerase Deficiency: High Prevalence of p.Arg347His Mutation in Indian Population Associated with Severe Hereditary Non-Spherocytic Hemolytic Anemia Coupled with Neurological Dysfunction

  • Prabhakar S. KedarEmail author
  • Rashmi Dongerdiye
  • Pooja Chilwirwar
  • Vinod Gupta
  • Ashish Chiddarwar
  • Rati Devendra
  • Prashant Warang
  • Harsha Prasada
  • Abhilasha Sampagar
  • Sunil Bhat
  • S. Chandrakala
  • Manisha Madkaikar
Original Article

Abstract

Objectives

Glucose-6-phosphate isomerase (GPI) deficiency is an autosomal recessive genetic disorder causing hereditary non-spherocytic hemolytic anemia (HNSHA) coupled with a neurological disorder. The aim of this study was to identify GPI genetic defects in a cohort of Indian patients with HNSHA coupled with neurological dysfunction.

Methods

Thirty-five patients were screened for GPI deficiency in the HNSHA patient group; some were having neurological dysfunction. Enzyme activity was measured by spectrophotometric method. The genetic study was done by single-stranded conformation polymorphism (SSCP) analysis, restriction fragment length polymorphism (RFLP) analysis by the restriction enzyme AciI for p.Arg347His (p.R347H) and confirmation by Sanger’s sequencing.

Results

Out of 35 patients, 15 showed 35% to 70% loss of GPI activity, leading to neurological problems with HNSHA. Genetic analysis of PCR products of exon 12 of the GPI gene showed altered mobility on SSCP gel. Sanger’s sequencing revealed a homozygous c1040G > A mutation predicting a p.Arg347His replacement which abolishes AciI restriction site. The molecular modeling analysis suggests p.Arg347 is involved in dimerization of the enzyme. Also, this mutation generates a more labile enzyme which alters its three-dimensional structure and function.

Conclusions

This report describes the high prevalence of p.Arg347His pathogenic variant identified in Indian GPI deficient patients with hemolytic anemia and neuromuscular impairment. It suggests that neuromuscular impairment with hemolytic anemia cases could be investigated for p.Arg347His pathogenic variant causing GPI deficiency because of neuroleukin activity present in the GPI monomer which has neuroleukin action at the same active site and generates neuromuscular problems as well as hemolytic anemia.

Keywords

Glucose-6-phosphate isomerase (GPI) deficiency Neurological dysfunction Hereditary non-spherocytic hemolytic anemia (HNSHA) Glycolysis Molecular modeling India 

Notes

Authors’ Contribution

PSK: Designed the experiment. RMD, AC, RD, PC, VG: Collected the biochemical, hematological data and performed lab experiments. HP, AS, SB, SC: Referred the case and did clinical examinations. PSK, PW, RD: Conducted the literature search, designed the figures and tables, and analyzed and interpreted the data. PSK, RD: Revised the first draft and completed the final manuscript. All authors approved the final manuscript. PC, VG, RD, AC, PW, PSK prepared the manuscript. MM is the Director, NIIH and also a clinician. She gave her suggestion about the cases and helped in clinical correlation; All authors read and approved the final manuscript. PSK is the guarantor.

Compliance with Ethical Standards

Conflict of Interest

None.

Source of Funding

This study was performed with the financial support from Indian Council of Medical Research, New Delhi, and Department of Biotechnology, New Delhi.

References

  1. 1.
    Manco L, Bento C, Victor BL, et al. Hereditary nonspherocytic hemolytic anemia caused by red cell glucose-6-phosphate isomerase (GPI) deficiency in two Portuguese patients: clinical features and molecular study. Blood Cells Mol Dis. 2016;60:18–23.CrossRefGoogle Scholar
  2. 2.
    Kugler W, Breme K, Laspe P, et al. Molecular basis of neurological dysfunction coupled with hemolytic anemia in human glucose-6-phosphate isomerase (GPI) deficiency. Hum Genet. 1998;103:450–4.CrossRefGoogle Scholar
  3. 3.
    Haga A, Niinaka Y, Raz A. Phosphohexose isomerase/autocrine motility factor/neuroleukin/maturation factor is a multifunctional phosphoprotein. Biochim Biophys Acta. 2000;1480:235–44.CrossRefGoogle Scholar
  4. 4.
    Ravindranath Y, Paglia DE, Warrier I, Valentine W, Nakatani M, Brockway RA. Glucose phosphate isomerase deficiency as a cause of hydrops fetalis. N Engl J Med. 1987;316:258–61.CrossRefGoogle Scholar
  5. 5.
    Kanno H, Fujii H, Hirono A, et al. Molecular analysis of glucose phosphate Isomerase deficiency associated with hereditary hemolytic anemia. Blood. 1996;88:2321–5.Google Scholar
  6. 6.
    Warang P, Kedar P, Ghosh K, Colah RB. Hereditary non-spherocytic hemolytic anemia and severe glucose phosphate isomerase deficiency in an Indian patient homozygous for the L487F mutation in the human GPI gene. Int J Hematol. 2012;96:263–7.CrossRefGoogle Scholar
  7. 7.
    Zaidi AU, Kedar P, Koduri PR, et al. Glucose phosphate isomerase (GPI) Tadikonda: characterization of a novel Pro340Ser mutation. Pediatr Hematol Oncol. 2017;34:449–54.CrossRefGoogle Scholar
  8. 8.
    Kedar P, Gupta V, Dongerdiye R, Chiddarwar A, Warang P, Madkaikar MR. Molecular diagnosis of unexplained hemolytic anemia using targeted next-generation sequencing panel revealed (p.Ala337Thr) novel mutation in the GPI gene in two Indian patients. J Clin Pathol. 2019;72:81–5.CrossRefGoogle Scholar
  9. 9.
    Walker JI, Layton DM, Bellingham AJ, Morgan MJ, Faik P. DNA sequence abnormalities in human glucose 6-phosphate isomerase deficiency. Hum Mol Genet. 1993;2:327–9.CrossRefGoogle Scholar
  10. 10.
    Kugler W, Lakomek M. Glucose-6-phosphate isomerase deficiency. Best Pract Res Clin Haematol. 2000;13:89–101.CrossRefGoogle Scholar
  11. 11.
    Beutler E, West C, Britton HA, Harris J, Forman L. Glucose phosphate isomerase (GPI) deficiency mutations associated with hereditary nonspherocytic hemolytic anemia (HNSHA). Blood Cells Mol Dis. 1997;23:402–9.CrossRefGoogle Scholar
  12. 12.
    Xu W, Beutler E. The characterization of gene mutations for human glucose phosphate isomerase deficiency associated with chronic hemolytic anemia. J Clin Invest. 1994;94:2326–9.CrossRefGoogle Scholar
  13. 13.
    Baronciani L, Zanella A, Bianchi P, et al. Study of the molecular defects in glucose phosphate isomerase-deficient patients affected by chronic hemolytic anemia. Blood. 1996;88:2306–10.Google Scholar
  14. 14.
    Repiso A, Oliva B, Vives-Corrons JL, Beutler E, Carreras J, Climent F. Red cell glucose phosphate isomerase (GPI): a molecular study of three novel mutations associated with hereditary nonspherocytic hemolytic anemia. Hum Mutat. 2006;27:1159.CrossRefGoogle Scholar
  15. 15.
    Clarke JL, Vulliamy TJ, Roper D. Combined glucose-6-phosphate dehydrogenase, and glucose phosphate isomerase deficiency can alter clinical outcome. Blood Cells Mol Dis. 2003;30:258–63.CrossRefGoogle Scholar
  16. 16.
    Lakomek M, Winkler H. Erythrocyte pyruvate kinase- and glucose phosphate isomerase deficiency: perturbation of glycolysis by structural defects and functional alterations of defective enzymes and its relation to the clinical severity of chronic hemolytic anemia. Biophys Chem. 1997;66:269–84.CrossRefGoogle Scholar
  17. 17.
    Mojzikova R, Koralkova P, Holub D, et al. Two novel mutations (p.(Ser160Pro) and p.(Arg472Cys)) causing glucose-6-phosphate isomerase deficiency are associated with erythroid dysplasia and inappropriately suppressed hepcidin. Blood Cells Mol Dis. 2018;69:23–9.CrossRefGoogle Scholar
  18. 18.
    Burger NCM, Van Wijk R, Bresters D, Schell EA. A novel mutation of glucose phosphate isomerase (GPI) causing severe neonatal anemia due to GPI deficiency. J Pediatr Hematol Oncol. 2018 Dec 21. [Ahead of print].Google Scholar
  19. 19.
    Zhu X, Petrovski S, Xie P, et al. Whole-exome sequencing in undiagnosed genetic diseases: interpreting 119 trios. Genet Med. 2015;17:774–81.CrossRefGoogle Scholar
  20. 20.
    Dacie JV, Lewis SM. Practical Haematology. 10th ed. Edinburgh: Churchill Livingstone; 2006. p. 60–78.Google Scholar
  21. 21.
    Beutler E. Red Cell Metabolism: A Manual of Biochemical Methods. 3rd ed. New York: Grune & Stratton, Inc; 1984.Google Scholar
  22. 22.
    Davies C, Muirhead H, Chirgwin J. The structure of human phosphoglucose isomerase complexed with a transition-state analog. Acta Crystallogr D Biol Crystallogr. 2003;59:1111–3.CrossRefGoogle Scholar
  23. 23.
    Feyfant E, Sali A, Fiser A. Modeling mutations in protein structures. Protein Sci. 2007;16:2030–41.CrossRefGoogle Scholar
  24. 24.
    Repiso A, Oliva B, Vives Corrons JL, Carreras J, Climent F. Glucose phosphate isomerase deficiency: enzymatic and familial characterization of Arg346His mutation. Biochim Biophys Acta. 2005;1740:467–71.CrossRefGoogle Scholar
  25. 25.
    Jamwal M, Aggarwal A, Das A, et al. Next-generation sequencing unravels homozygous mutation in glucose-6-phosphate isomerase, GPIc.1040G> A (p.Arg347His) causing hemolysis in an Indian infant. Clin Chim Acta. 2017;468:81–4.CrossRefGoogle Scholar
  26. 26.
    Magor GW, Tallack MR, Gillinder KR, et al. KLF1-null neonates display hydrops fetalis and a deranged erythroid transcriptome. Blood. 2015;125:2405–17.CrossRefGoogle Scholar
  27. 27.
    Koralkova P, van Solinge WW, van Wijk R. Rare hereditary red blood cell enzymopathies associated with hemolytic anemia pathophysiology, clinical aspects, and laboratory diagnosis. Int J Lab Hematol. 2014;36:388–97.CrossRefGoogle Scholar
  28. 28.
    Grace RF, Zanella A, Neufeld EJ, et al. Erythrocyte pyruvate kinase deficiency: 2015 status report. Am J Hematol. 2015;90:825–30.CrossRefGoogle Scholar
  29. 29.
    Shalev O, Shalev RS, Forman L, Beutler E. GPI mount scopus--a variant of glucose phosphate isomerase deficiency. Ann Hematol. 1993;67:197–200.CrossRefGoogle Scholar
  30. 30.
    Schröter W, Eber SW, Bardosi A, Gahr M, Gabriel M, Sitzmann FC. Generalized glucose phosphate Isomerase (GPI) deficiency causing hemolytic anemia, neuromuscular symptoms and impairment of granulocytic function: a new syndrome due to a new stable GPI variant with a diminished specific activity (GPI Homburg). Eur J Pediatr. 1985;144:301–5.CrossRefGoogle Scholar
  31. 31.
    Roy NB, Wilson EA, Henderson S, et al. A novel 33-gene-targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rarely inherited anemias. Br J Haematol. 2016;175:318–30.CrossRefGoogle Scholar
  32. 32.
    Neubauer BA, Eber SW, Lakomek M, Gahr M, Schröter W. Combination of congenital nonspherocytic hemolytic anemia and impairment of granulocyte function in severe glucose phosphate isomerse deficiency. A new variant enzyme designated GPI Calden. Acta Haematol. 1990;83:206–10.CrossRefGoogle Scholar

Copyright information

© Dr. K C Chaudhuri Foundation 2019

Authors and Affiliations

  • Prabhakar S. Kedar
    • 1
    Email author
  • Rashmi Dongerdiye
    • 1
  • Pooja Chilwirwar
    • 1
  • Vinod Gupta
    • 1
  • Ashish Chiddarwar
    • 1
  • Rati Devendra
    • 1
  • Prashant Warang
    • 1
  • Harsha Prasada
    • 2
  • Abhilasha Sampagar
    • 3
  • Sunil Bhat
    • 4
  • S. Chandrakala
    • 5
  • Manisha Madkaikar
    • 1
  1. 1.Department of HematogeneticsICMR-National Institute of Immunohematology, King Edward Memorial Hospital CampusParelIndia
  2. 2.Department of Pediatrics, Kasturba Medical College HospitalManipal UniversityMangaloreIndia
  3. 3.Department of PediatricsKLES Dr. Prabhakar Kore Hospital, and MRCBelagaviIndia
  4. 4.Department of Hematology, Oncology, and Bone Marrow TransplantationMazumdar Shaw Cancer CenterBangaloreIndia
  5. 5.Department of HematologyKing Edward Memorial HospitalParelIndia

Personalised recommendations