Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis
An erythrocytosis is present when the red blood cell mass is increased, demonstrated as elevated hemoglobin and hematocrit in the laboratory evaluation. Congenital predispositions for erythrocytosis are rare, with germline variants in several genes involved in oxygen sensing (VHL, EGLN1, and EPAS1),...
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Frontiers Media S.A.
2021-07-01
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Online Access: | https://www.frontiersin.org/articles/10.3389/fgene.2021.689868/full |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Aleša Kristan Tadej Pajič Tadej Pajič Tadej Pajič Aleš Maver Tadeja Režen Tanja Kunej Rok Količ Andrej Vuga Martina Fink Martina Fink Špela Žula Helena Podgornik Helena Podgornik Saša Anžej Doma Saša Anžej Doma Irena Preložnik Zupan Irena Preložnik Zupan Damjana Rozman Nataša Debeljak |
spellingShingle |
Aleša Kristan Tadej Pajič Tadej Pajič Tadej Pajič Aleš Maver Tadeja Režen Tanja Kunej Rok Količ Andrej Vuga Martina Fink Martina Fink Špela Žula Helena Podgornik Helena Podgornik Saša Anžej Doma Saša Anžej Doma Irena Preložnik Zupan Irena Preložnik Zupan Damjana Rozman Nataša Debeljak Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis Frontiers in Genetics NGS – next-generation sequencing erythrocytosis targeted panel sequencing iron metabolism diagnostics rare disease (RD) |
author_facet |
Aleša Kristan Tadej Pajič Tadej Pajič Tadej Pajič Aleš Maver Tadeja Režen Tanja Kunej Rok Količ Andrej Vuga Martina Fink Martina Fink Špela Žula Helena Podgornik Helena Podgornik Saša Anžej Doma Saša Anžej Doma Irena Preložnik Zupan Irena Preložnik Zupan Damjana Rozman Nataša Debeljak |
author_sort |
Aleša Kristan |
title |
Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis |
title_short |
Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis |
title_full |
Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis |
title_fullStr |
Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis |
title_full_unstemmed |
Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing Analysis |
title_sort |
identification of variants associated with rare hematological disorder erythrocytosis using targeted next-generation sequencing analysis |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Genetics |
issn |
1664-8021 |
publishDate |
2021-07-01 |
description |
An erythrocytosis is present when the red blood cell mass is increased, demonstrated as elevated hemoglobin and hematocrit in the laboratory evaluation. Congenital predispositions for erythrocytosis are rare, with germline variants in several genes involved in oxygen sensing (VHL, EGLN1, and EPAS1), signaling for hematopoietic cell maturation (EPOR and EPO), and oxygen transfer (HBB, HBA1, HBA2, and BPGM) that were already associated with the eight congenital types (ECYT1–8). Screening for variants in known congenital erythrocytosis genes with classical sequencing approach gives a correct diagnosis for only up to one-third of the patients. The genetic background of erythrocytosis is more heterogeneous, and additional genes involved in erythropoiesis and iron metabolism could have a putative effect on the development of erythrocytosis. This study aimed to detect variants in patients with yet unexplained erythrocytosis using the next-generation sequencing (NGS) approach, targeting genes associated with erythrocytosis and increased iron uptake and implementing the diagnostics of congenital erythrocytosis in Slovenia. Selected 25 patients with high hemoglobin, high hematocrit, and no acquired causes were screened for variants in the 39 candidate genes. We identified one pathogenic variant in EPAS1 gene and three novel variants with yet unknown significance in genes EPAS1, JAK2, and SH2B3. Interestingly, a high proportion of patients were heterozygous carriers for two variants in HFE gene, otherwise pathogenic for the condition of iron overload. The association between the HFE variants and the development of erythrocytosis is not clearly understood. With a targeted NGS approach, we determined an actual genetic cause for the erythrocytosis in one patient and contributed to better management of the disease for the patient and his family. The effect of variants of unknown significance on the enhanced production of red blood cells needs to be further explored with functional analysis. This study is of great significance for the improvement of diagnosis of Slovenian patients with unexplained erythrocytosis and future research on the etiology of this rare hematological disorder. |
topic |
NGS – next-generation sequencing erythrocytosis targeted panel sequencing iron metabolism diagnostics rare disease (RD) |
url |
https://www.frontiersin.org/articles/10.3389/fgene.2021.689868/full |
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doaj-e2d1e10e99874627bc2743516f61c7582021-07-19T12:55:37ZengFrontiers Media S.A.Frontiers in Genetics1664-80212021-07-011210.3389/fgene.2021.689868689868Identification of Variants Associated With Rare Hematological Disorder Erythrocytosis Using Targeted Next-Generation Sequencing AnalysisAleša Kristan0Tadej Pajič1Tadej Pajič2Tadej Pajič3Aleš Maver4Tadeja Režen5Tanja Kunej6Rok Količ7Andrej Vuga8Martina Fink9Martina Fink10Špela Žula11Helena Podgornik12Helena Podgornik13Saša Anžej Doma14Saša Anžej Doma15Irena Preložnik Zupan16Irena Preložnik Zupan17Damjana Rozman18Nataša Debeljak19Medical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaDepartment of Hematology, University Medical Centre Ljubljana, Ljubljana, SloveniaClinical Institute of Genomic Medicine, University Medical Centre Ljubljana, Ljubljana, SloveniaClinical Biochemistry, Faculty of Medicine, University of Maribor, Maribor, SloveniaClinical Institute of Genomic Medicine, University Medical Centre Ljubljana, Ljubljana, SloveniaCentre for Functional Genomics and Bio-Chips, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaDepartment of Animal Science, Biotechnical Faculty, University of Ljubljana, Ljubljana, SloveniaKemomed Research and Development, Kemomed Ltd., Ljubljana, SloveniaKemomed Research and Development, Kemomed Ltd., Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaDepartment of Hematology, University Medical Centre Ljubljana, Ljubljana, SloveniaDepartment of Hematology, University Medical Centre Ljubljana, Ljubljana, SloveniaDepartment of Hematology, University Medical Centre Ljubljana, Ljubljana, SloveniaClinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, Ljubljana, SloveniaDepartment of Hematology, University Medical Centre Ljubljana, Ljubljana, SloveniaDepartment of Internal Medicine, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaDepartment of Hematology, University Medical Centre Ljubljana, Ljubljana, SloveniaDepartment of Internal Medicine, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaCentre for Functional Genomics and Bio-Chips, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaAn erythrocytosis is present when the red blood cell mass is increased, demonstrated as elevated hemoglobin and hematocrit in the laboratory evaluation. Congenital predispositions for erythrocytosis are rare, with germline variants in several genes involved in oxygen sensing (VHL, EGLN1, and EPAS1), signaling for hematopoietic cell maturation (EPOR and EPO), and oxygen transfer (HBB, HBA1, HBA2, and BPGM) that were already associated with the eight congenital types (ECYT1–8). Screening for variants in known congenital erythrocytosis genes with classical sequencing approach gives a correct diagnosis for only up to one-third of the patients. The genetic background of erythrocytosis is more heterogeneous, and additional genes involved in erythropoiesis and iron metabolism could have a putative effect on the development of erythrocytosis. This study aimed to detect variants in patients with yet unexplained erythrocytosis using the next-generation sequencing (NGS) approach, targeting genes associated with erythrocytosis and increased iron uptake and implementing the diagnostics of congenital erythrocytosis in Slovenia. Selected 25 patients with high hemoglobin, high hematocrit, and no acquired causes were screened for variants in the 39 candidate genes. We identified one pathogenic variant in EPAS1 gene and three novel variants with yet unknown significance in genes EPAS1, JAK2, and SH2B3. Interestingly, a high proportion of patients were heterozygous carriers for two variants in HFE gene, otherwise pathogenic for the condition of iron overload. The association between the HFE variants and the development of erythrocytosis is not clearly understood. With a targeted NGS approach, we determined an actual genetic cause for the erythrocytosis in one patient and contributed to better management of the disease for the patient and his family. The effect of variants of unknown significance on the enhanced production of red blood cells needs to be further explored with functional analysis. This study is of great significance for the improvement of diagnosis of Slovenian patients with unexplained erythrocytosis and future research on the etiology of this rare hematological disorder.https://www.frontiersin.org/articles/10.3389/fgene.2021.689868/fullNGS – next-generation sequencingerythrocytosistargeted panel sequencingiron metabolismdiagnosticsrare disease (RD) |