Skip to main content
SpringerLink
Log in

Red cell ektacytometry in two patients with chronic hemolytic anemia and three new α-spectrin variants

  • Original Article
  • Published:
Annals of Hematology Aims and scope Submit manuscript

Abstract

Red blood cell (RBC) morphology is, in general, the key diagnostic feature for hereditary spherocytosis (HS) and hereditary elliptocytosis (HE). However, in hereditary pyropoikilocytosis (HPP), the severe clinical form of HE, the morphological diagnosis is difficult due to the presence of a RBC morphological picture characterized by a mixture of elliptocytes, spherocytes, tear-drop cells, and fragmented cells. This difficulty increases in new-borns and/or patients requiring frequent transfusions, making impossible the prediction of the disease course or its severity. Recently, it has been demonstrated that the measurement of osmotic gradient ektacytometry (OGE), using a laser-assisted optical rotational ektacytometer LoRRca (MaxSis, RR Mechatronics), allows a clear differentiation between HS and HE, where the truncated osmoscan curve reflects the inability of the already elliptical cells to deform further under shear stress in the face of hypotonicity. In HPP, however, the RBCs appear to have a significantly decreased ability to maintain deformability in these conditions, and the classical trapezoidal profile of HE is less evident or indistinguishable from HS. Here, two unrelated patients with hereditary hemolytic anemia (HHA) due to HPP and HS, respectively, are described with the joint inheritance of a complex set of five genetic defects. Two of these defects are novel alpha-spectrin gene (SPTA1) variants, one is a microdeletion that removes the entire SPTA1 gene, and two are well-known low-expression polymorphic alleles: α-LELY and α-LEPRA. In the HPP patient (ID1), with many circulating spherocytes, the interactions between the two SPTA1 gene variants may lead, in addition to an elongation defect (elliptocytes), to a loss of membrane stability and vesiculation (spherocytes), and RBCs appear to have a significantly decreased ability to maintain deformability in hypotonic conditions. Due to this, the classical trapezoidal profile of HE may become less evident or indistinguishable from HS. The second patient (ID2) was a classical severe form of HS with the presence of more than 20% of spherocytes and few pincered cells. The severity of clinical manifestation is due to the coinheritance of a microdeletion of chromosome 1 that removes the entire SPTA1 gene with a LEPRA SPTA1 variant in trans. The diagnostic interest of both observations is discussed.

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

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Iolascon A, Andolfo I, Russo R (2019) Advances in understanding the pathogenesis of red cell membrane disorders. Br J Haematol 187(1):13–24

    Article  Google Scholar 

  2. Barcellini W, Fattizzo B (2015) Clinical applications of hemolytic markers in the differential diagnosis and management of hemolytic anemia. Dis Markers 2015:635670. https://doi.org/10.1155/2015/63567

  3. Narla J, Mohandas N (2017) Red cell membrane disorders. Int J Lab Hematol 39:47–52

    Article  Google Scholar 

  4. Fermo E, Vercellati C, Bianchi P (2021) Screening tools for hereditary hemolytic anemia: new concepts and strategies. Expert Rev Hematol 14(3):281–292

    Article  CAS  Google Scholar 

  5. Cochran DL, Burnside LK (1999) Detecting and identifying hereditary pyropoikilocytosis. Lab Med 30(1):26–29

    Article  Google Scholar 

  6. Fermo E, Vercellati C, Marcello AP, Keskin EY, Perrotta S, Zaninoni A, ... Bianchi P (2021) Targeted next generation sequencing and diagnosis of congenital hemolytic anemias: a three years experience monocentric study. Front Physiol 12:684569

  7. Zaidi AU, Buck S, Gadgeel M, Herrera-Martinez M, Mohan A, Johnson K, ... Ravindranath Y (2020) Clinical diagnosis of red cell membrane disorders: comparison of osmotic gradient ektacytometry and eosin maleimide (EMA) fluorescence test for red cell band 3 (AE1, SLC4A1) content for clinical diagnosis. Front Physiol 11:636. https://doi.org/10.3389/fphys.2020.00636

  8. Russo R AI, Manna F, Gambale A, Pignataro P, De Rosa G, Iolascon A (2016) RedPlex: a targeted next-generation sequencing-based diagnosis for patients with hereditary haemolytic anemias. EMJ Hematol 6(1):50–52

  9. Vives-Corrons JL, Krishnevskaya E, Rodriguez IH, Ancochea A (2021) Characterization of hereditary red blood cell membranopathies using combined targeted next-generation sequencing and osmotic gradient ektacytometry. Int J Hematol 113(2):163–174

    Article  CAS  Google Scholar 

  10. Llaudet-Planas E, Vives-Corrons JL, Rizzuto V, Gómez-Ramírez P, Sevilla Navarro J, CollSibina MT et al (2018) Osmotic gradient ektacytometry: a valuable screening test for hereditary spherocytosis and other red blood cell membrane disorders. Int J Lab Hem 40:94–102

    Article  CAS  Google Scholar 

  11. Lux SE (2016) Anatomy of the red cell membrane skeleton: unanswered questions. Blood 127(2):187–199

    Article  CAS  Google Scholar 

  12. Speicher DW, Weglarz L, DeSilva TM (1992) Properties of human red cell spectrin heterodimer (side-to-side) assembly and identification of an essential nucleation site. J Biol Chem 267(21):14775–14782

    Article  CAS  Google Scholar 

  13. An X, Lecomte MC, Chasis JA, Mohandas N, Gratzer W (2002) Shear-response of the spectrin dimer-tetramer equilibrium in the red blood cell membrane. J Biol Chem 277(35):31796–31800

    Article  CAS  Google Scholar 

  14. Liu SC, Palek J, Prchal J, Castleberry RP (1981) Altered spectrin dimer-dimer association and instability of erythrocyte membrane skeletons in hereditary pyropoikilocytosis. J Clin Investig 68(3):597–605

    Article  CAS  Google Scholar 

  15. Gaetani M, Mootien S, Harper S, Gallagher PG, Speicher DW (2008) Structural and functional effects of hereditary hemolytic anemia-associated point mutations in the alpha spectrin tetramer site. Blood, J Am Soc Hemat 111(12):5712–5720

    CAS  Google Scholar 

  16. Da Costa L, Galimand J, Fenneteau O, Mohandas N (2013) Hereditary spherocytosis, elliptocytosis, and other red cell membrane disorders. Blood Rev 27(4):167–178

    Article  Google Scholar 

  17. Agre P (2020) Partial deficiencies of erythrocyte spectrin in hereditary spherocytosis. In Red Blood Cell Membranes (pp. 707–726). CRC Press

  18. Delaunay J, Nouyrigat V, Proust A, Schischmanoff PO, Cynober T, Yvart J... & Tchernia G (2004) Different impacts of alleles αLEPRA and αLELY as assessed versus a novel, virtually null allele of the SPTA1 gene in trans. British J haem, 127(1), 118-122

  19. Wichterle H, Hanspal M, Palek J, Jarolim P (1996) Combination of two mutant alpha spectrin alleles underlies a severe spherocytic hemolytic anemia. J Clin Investig 98(10):2300–2307

    Article  CAS  Google Scholar 

  20. Wilmotte R, Mare’chal J, Morle’ L, Baklouti F, Philippe N, Kastally R, Kotula L, Delaunay J, Alloisio N (1993) Low expression allele aLELY of red cell spectrin is associated with mutations in exon 40 (aV/41 polymorphism) and intron 45 and with partial skipping of exon 46. J Clin Invest 91:2091–2096

    Article  CAS  Google Scholar 

  21. Alloisio N, Morle’ L, Mare’chal J, Roux AF, Ducluzeau MT, Guetarni D, Pothier B, Baklouti F, Ghanem A, Kastally R, Delaunay J (1991) SpaV/41: a common spectrin polymorphism at the aIV-aV domain junction. Relevance to the expression level of hereditary elliptocytosis due to a-spectrin variants located in trans. J Clin Invest 87:2169–2177

    Article  CAS  Google Scholar 

  22. Wilmotte R, Harper SL, Ursitti JA, Maréchal J, Delaunay J, Speicher DW (1997) The exon 46-encoded sequence is essential for stability of human erythroid α-spectrin and heterodimer formation. Blood, J Am Soc Hem 90(10):4188–4196

    CAS  Google Scholar 

  23. Gallagher PG, Maksimova Y, Lezon-Geyda K, Newburger PE, Medeiros D, Hanson RD, ... & Schulz VP (2019) Aberrant splicing contributes to severe α-spectrin–linked congenital hemolytic anemia. J Clin Invest, 129(7), 2878-2887

  24. Knowles WJ, Morrow JS, Speicher DW et al (1983) Molecular and functional changes in spectrin from patients with hereditary pyropoikilocytosis. J Clin Investig 71(6):1867–1877. https://doi.org/10.1172/jci110942

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Larocci TA, Wagner GM, Mohandas N, Lane PA, Mentzer WC (1988) Hereditary poikilocytic anemia associated with the co-inheritance of two alpha spectrin abnormalities. Blood 71:1390–1396

    Article  Google Scholar 

  26. Mohandas N, Clark MR, Health BP, Rossi M, Wolfe LC, Lux SE, Shohet SB (1982) A technique to detect reduced mechanical stability of red cell membranes: relevance to elliptocytic disorders. Blood. 59:768–774

    Article  CAS  Google Scholar 

  27. Hanspal M, Hanspal JS, Sahr KE, Fibach E, Nachman J, Palek J (1993) Molecular basis of spectrin deficiency in hereditary pyropoikilocytosis. Blood 82(5):1652–1660 (PMID: 8364214)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are indebted to the European Commission for the Equality Plus (Erasmus +) Grant (Ref. 2019-1-TR01-KA202-076789)

Author information

Authors and Affiliations

  1. Red Cell Pathology and Haematopoietic Disorders (Rare Anaemias Unit), Institute for Leukaemia Research Josep Carreras (IJC), Ctra de Can Ruti, Camí de les Escoles s/n Badalona, 08916, Barcelona, Spain

    Joan-Lluis Vives-Corrons & Elena Krishnevskaya

  2. Haematology Department, Hospital Universitari Germans Trias I Pujol, Badalona, Barcelona, Spain

    Inés Hernández-Rodriguez

  3. Red Blood Cell Disorders Unit, Hematology Department, Virgen del Rocío University Hospital, Institute of Biomedicine of Seville (IBiS-CSIC), Seville, Spain

    Salvador Payán-Pernia

  4. Hematology Department, Hospital Universitari de La Santa Creu i Sant Pau, Barcelona, Spain

    Ángel F. Remacha Sevilla & Isabel Badell

  5. Department of Pediatrics, Hospital Universitari de La Santa Creu i Sant Pau, National Reference Center (CSUR Accreditation) for Hereditary Red Blood Cell Disorders (Hospital de La Santa Creu i Sant Pau-Hospital Sant Joan de Déu), Barcelona, Spain

    Isabel Badell

Authors
  1. Joan-Lluis Vives-Corrons
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elena Krishnevskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Inés Hernández-Rodriguez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Salvador Payán-Pernia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ángel F. Remacha Sevilla
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Isabel Badell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Joan-Lluis Vives-Corrons.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Research involving human participants and/or animals

All procedures performed in studies 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.

Informed consent

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

Disclosure

The authors have no other relevant affiliations or financial involvement with any organisation or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Vives-Corrons, JL., Krishnevskaya, E., Hernández-Rodriguez, I. et al. Red cell ektacytometry in two patients with chronic hemolytic anemia and three new α-spectrin variants. Ann Hematol 101, 549–555 (2022). https://doi.org/10.1007/s00277-021-04723-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00277-021-04723-5

Keywords

Search

Navigation