244c6058e0921bc8c9d0e225cf68ad07 270643.pdf 49b67159a50e16a7763518dc5696efe05948aedc 270643.pdf be4398a043ff74d2a0b240b6c0c06c5f38b7016975af2a5a5ff6f49d35b6c002 270643.pdf Title: Concomitant Hereditary Spherocytosis and Pyruvate Kinase Deficiency in a Spanish Family with Chronic Hemolytic Anemia: Contribution of Laser Ektacytometry to Clinical Diagnosis Subject: 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 -spectrin (SPTB) c.647G>A variant and pyruvate kinase (PKLR) c.1706G>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 -spectrin (SPTB; c.647G>A) in seven members with HS, three of them co-inherited the PKLR variant c.1706G>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>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. Keywords: anemia; hemolysis; RBC membrane; pyruvate-kinase deficiency; beta-spectrin; hereditary spherocytosis Author: Joan-Lluis Vives Corrons, Elena Krishnevskaya, Laura Montllor, Valentina Leguizamon and Marta Garcia Bernal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Mar 28 09:17:49 2022 CEST ModDate: Mon Mar 28 09:22:04 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 8 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 958564 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- DRZUJM+VnURWPalladioL Type 1 Custom yes yes no 10 0 QDCWHF+EURM10 Type 1 Builtin yes yes no 15 0 WNPGHG+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 EEMDES+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 NLRDGI+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 36 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 40 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 44 0 PFKOHU+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 56 0 AINHDX+CMSY10 Type 1 Builtin yes yes no 81 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 87 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 93 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 99 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 111 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 117 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 123 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 133 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 139 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 145 0 FNTSBS+PalatinoLinotype-BoldItalic CID TrueType Identity-H yes yes yes 151 0 CRUZTT+VnURWPalladioL-Italic Type 1 Custom yes yes no 168 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-05-30 02:32:11 CEST RepresentationInformation: 270643.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-05-29 11:56:15 CEST Size: 958564 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 293 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Info: Title: Concomitant Hereditary Spherocytosis and Pyruvate Kinase Deficiency in a Spanish Family with Chronic Hemolytic Anemia: Contribution of Laser Ektacytometry to Clinical Diagnosis Author: Joan-Lluis Vives Corrons, Elena Krishnevskaya, Laura Montllor, Valentina Leguizamon and Marta Garcia Bernal Subject: 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 -spectrin (SPTB) c.647G>A variant and pyruvate kinase (PKLR) c.1706G>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 -spectrin (SPTB; c.647G>A) in seven members with HS, three of them co-inherited the PKLR variant c.1706G>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>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. 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