{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:57:28Z","timestamp":1760151448139,"version":"build-2065373602"},"reference-count":35,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,3,28]],"date-time":"2022-03-28T00:00:00Z","timestamp":1648425600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Background: Hereditary spherocytosis (HS) and pyruvate kinase deficiency (PKD) are the most common causes of hereditary chronic hemolytic anemia. Here, we describe clinical and genetic characteristics of a Spanish family with concomitant \u03b2-spectrin (SPTB) c.647G&gt;A variant and pyruvate kinase (PKLR) c.1706G&gt;A variant. Methods: A family of 11 members was studied. Hematological investigation, hemolysis tests, and specific red cell studies were performed in all family members, according to conventional procedures. An ektacytometric study was performed using the osmoscan module of the Lorca ektacytometer (MaxSis. RR Mechatronics). The presence of the SPTB and PKLR variants was confirmed by t-NGS. Results: The t-NGS genetic characterization of the 11 family members showed the presence of a heterozygous mutation for the \u03b2-spectrin (SPTB; c.647G&gt;A) in seven members with HS, three of them co-inherited the PKLR variant c.1706G&gt;A. In the remaining four members, no gene mutation was found. Ektacytometry allowed a clear diagnostic orientation of HS, independently from the PKLR variant. Conclusions: This family study allows concluding that the SPTB mutation, (c.647G&gt;A) previously described as likely pathogenic (LP), should be classified as pathogenic (P), according to the recommendations for pathogenicity of the American College of Medical Genetics and the Association for Molecular Pathology. In addition, after 6 years of clinical follow-up of the patients with HS, it can be inferred that the chronic hemolytic anemia may be attributable to the SPTB mutation only, without influence of the concomitant PKLR. Moreover, only the family members with the SPTB mutation exhibited an ektacytometric profile characteristic of HS.<\/jats:p>","DOI":"10.3390\/cells11071133","type":"journal-article","created":{"date-parts":[[2022,3,29]],"date-time":"2022-03-29T21:40:14Z","timestamp":1648590014000},"page":"1133","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Concomitant Hereditary Spherocytosis and Pyruvate Kinase Deficiency in a Spanish Family with Chronic Hemolytic Anemia: Contribution of Laser Ektacytometry to Clinical Diagnosis"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1820-2594","authenticated-orcid":false,"given":"Joan-Lluis","family":"Vives Corrons","sequence":"first","affiliation":[{"name":"Red Cell Pathology and Hematopoietic Disorders, Institute for Leukaemia Research Josep Carreras, 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elena","family":"Krishnevskaya","sequence":"additional","affiliation":[{"name":"Red Cell Pathology and Hematopoietic Disorders, Institute for Leukaemia Research Josep Carreras, 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Montllor","sequence":"additional","affiliation":[{"name":"Red Cell Pathology and Hematopoietic Disorders, Institute for Leukaemia Research Josep Carreras, 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentina","family":"Leguizamon","sequence":"additional","affiliation":[{"name":"Red Cell Pathology and Hematopoietic Disorders, Institute for Leukaemia Research Josep Carreras, 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta","family":"Garcia Bernal","sequence":"additional","affiliation":[{"name":"Pediatric Hematology Department, University Hospital M\u00fatua Terrassa Terrassa, 08221 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1111\/j.1365-2141.1999.01130.x","article-title":"Red blood cell membrane disorders","volume":"104","author":"Tse","year":"1999","journal-title":"Br. J. Haematol."},{"key":"ref_2","unstructured":"Sans-Sabrafen, J., Besses Raebel, C., and Vives Corrons, J.L. (2006). Anemias hemol\u00edticas. Fisiopatolog\u00eda y diagn\u00f3stico. Anemias hemol\u00edticas. cong\u00e9nitas. Hematolog\u00eda Cl\u00ednica, Elsevier. [5th ed.]."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e24034","DOI":"10.1002\/jcla.24034","article-title":"The diagnostic protocol for hereditary spherocytosis-2021 update","volume":"35","author":"Wu","year":"2021","journal-title":"J. Clin. Lab. Anal."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1111\/ijlh.12746","article-title":"Osmotic gradient ektacytometry: A valuable screening test for hereditary spherocytosis and other red blood cell membrane disorders","volume":"40","author":"Rizzuto","year":"2018","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s12185-020-03010-9","article-title":"Characterization of hereditary red blood cell membranopathies using combined targeted next-generation sequencing and osmotic gradient ektacytometry","volume":"113","author":"Krishnevskaya","year":"2021","journal-title":"Int. J. Hematol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1002\/ajh.2830400207","article-title":"Splenic sequestration associated with sickle cell trait and hereditary spherocytosis","volume":"40","author":"Yang","year":"1992","journal-title":"Am. J. Hematol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1111\/j.1365-2141.1993.tb03347.x","article-title":"Coexistence of hereditary spherocytosis (HS) due to band 3 deficiency and \u03b2-thalassaemia trait: Partial correction of HS phenotype","volume":"85","author":"Perrotta","year":"1993","journal-title":"Br. J. Haematol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1111\/j.1365-2141.1994.tb04885.x","article-title":"Glucose 6-phosphate dehydrogenase deficiency and red cell membrane defects: Additive or synergistic interaction in producing chronic haemolytic anaemia","volume":"87","author":"Alfinito","year":"1994","journal-title":"Br. J. Haematol."},{"key":"ref_9","first-page":"176","article-title":"Combination of pyruvate kinase (PK) deficiency and hereditary spherocytosis (HS)","volume":"18","author":"Brook","year":"1970","journal-title":"Clin. Res."},{"key":"ref_10","first-page":"227","article-title":"Co-existence of hereditary spherocytosis and a new red cell pyruvate kinase variant: PK Mallorca","volume":"85","author":"Zarza","year":"2000","journal-title":"Haematologica"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"421","DOI":"10.7754\/Clin.Lab.2012.120905","article-title":"A case of hereditary spherocytosis misdiagnosed as pyruvate kinase deficient hemolytic anemia","volume":"59","author":"Vercellati","year":"2013","journal-title":"Clin. Lab."},{"key":"ref_12","first-page":"E35","article-title":"Partial pyruvate kinase deficiency aggravates the phenotypic expression of band 3 deficiency in a family with hereditary spherocytosis","volume":"90","author":"Veldthuis","year":"2015","journal-title":"Am. J. Hematol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1111\/bjh.16084","article-title":"Hereditary spherocytosis is associated with decreased pyruvate kinase activity due to impaired structural integrity of the red blood cell membrane","volume":"187","author":"Andres","year":"2019","journal-title":"Br. J. Haematol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3585","DOI":"10.1182\/blood.V95.11.3585","article-title":"Estimating the prevalence of pyruvate kinase deficiency from the gene frequency in the general white population","volume":"95","author":"Beutler","year":"2000","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1111\/ejh.13424","article-title":"Prevalence of pyruvate kinase deficiency: A systematic literature review","volume":"105","author":"Secrest","year":"2020","journal-title":"Eur. J. Haematol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2183","DOI":"10.1182\/blood-2017-10-810796","article-title":"Clinical spectrum of pyruvate kinase deficiency: Data from the pyruvate kinase deficiency natural history study","volume":"131","author":"Grace","year":"2018","journal-title":"Blood"},{"key":"ref_17","unstructured":"Enegela, O.A., and Anjum, F. (2021). Pyruvate Kinase deficiency. Stat Pearls, Stat Pearls Publishing."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1007\/BF00287063","article-title":"Hereditary erythrocyte pyruvate-kinase (PK) deficiency and chronic hemolytic anemia: Clinical, genetic and molecular studies in six new Spanish patients","volume":"53","author":"Marie","year":"1980","journal-title":"Hum. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1046\/j.1365-2141.1998.01013.x","article-title":"Molecular characterisation of PK-LR gene in pyruvate kinase-deficient patients","volume":"103","author":"Zarza","year":"1998","journal-title":"Br. J. Haematol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.medcli.2016.10.004","article-title":"Red cell pyruvate kinase deficiency in Spain: A study of 15 cases","volume":"148","author":"Montllor","year":"2017","journal-title":"Med. Clin."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e276","DOI":"10.1097\/HS9.0000000000000276","article-title":"The Complexity of Genotype-Phenotype Correlations in Hereditary Spherocytosis: A Cohort of 95 Patients: Genotype-Phenotype Correlation in Hereditary Spherocytosis","volume":"3","author":"Huisjes","year":"2019","journal-title":"Hemasphere"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and guidelines for the interpretation of sequence variants: A joint consensus recommendation of the American College of Medical Genetics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"ref_23","unstructured":"Vives Corrons, J.L., and Aguilar Bascompte, J.L. (2014). Manual of Hematology Laboratory Diagnostic Procedures, Elsevier. [4th ed.]. (In Spanish)."},{"key":"ref_24","unstructured":"Beutler, E. (1984). Red Cell Metabolism: A Manual of Biochemical Methods, Grune & Stratton, Inc."},{"key":"ref_25","first-page":"924","article-title":"Rapid flow cytometric test for the diagnosis of membrane cytoskeleton-associated haemolytic anaemia","volume":"111","author":"King","year":"2000","journal-title":"Br. J. Haematol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1111\/ijlh.12070","article-title":"Hereditary red cell membrane disorders and laboratory diagnostic testing","volume":"35","author":"King","year":"2013","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1111\/j.1365-2141.2005.05520.x","article-title":"Red cell pyruvate kinase deficiency: 17 new mutations of the PK-LR gene","volume":"129","author":"Fermo","year":"2005","journal-title":"Br. J. Haematol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1111\/j.1600-0609.1981.tb00448.x","article-title":"High prevalence of increased osmotic fragility of red blood cells among Norwegian blood donors","volume":"27","author":"Godal","year":"1981","journal-title":"Scand. J. Haematol."},{"key":"ref_29","first-page":"994","article-title":"Erythroid membrane protein defects in hereditary spherocytosis. A study of 62 Spanish cases","volume":"85","author":"Ricard","year":"2000","journal-title":"Haematologica"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1310","DOI":"10.3324\/haematol.12546","article-title":"Clinical and hematologic features of 300 patients affected by hereditary spherocytosis grouped according to the type of the membrane protein defect","volume":"93","author":"Mariani","year":"2008","journal-title":"Haematologica"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"516","DOI":"10.3324\/haematol.2011.052845","article-title":"Diagnostic power of laboratory tests for hereditary spherocytosis: A comparison study in 150 patients grouped according to molecular and clinical characteristics","volume":"97","author":"Bianchi","year":"2012","journal-title":"Haematologica"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1016\/S0140-6736(08)61588-3","article-title":"Hereditary spherocytosis","volume":"18","author":"Perrotta","year":"2008","journal-title":"Lancet"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1002\/ajh.24088","article-title":"Erythrocyte pyruvate kinase deficiency: 2015 status report","volume":"90","author":"Grace","year":"2015","journal-title":"Am. J. Hematol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2218","DOI":"10.3324\/haematol.2019.241141","article-title":"Molecular heterogeneity of pyruvate kinase deficiency","volume":"105","author":"Bianchi","year":"2020","journal-title":"Haematologica"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1186\/s13023-021-02036-4","article-title":"Confounding factors in the diagnosis and clinical course of rare congenital hemolytic anemias","volume":"16","author":"Fattizzo","year":"2021","journal-title":"Orphanet J. Rare Dis."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/7\/1133\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:44:40Z","timestamp":1760136280000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/7\/1133"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,28]]},"references-count":35,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,4]]}},"alternative-id":["cells11071133"],"URL":"https:\/\/doi.org\/10.3390\/cells11071133","relation":{},"ISSN":["2073-4409"],"issn-type":[{"type":"electronic","value":"2073-4409"}],"subject":[],"published":{"date-parts":[[2022,3,28]]}}}