Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia
Background Chronic lymphocytic leukemia (CLL) is a highly genetically heterogeneous disease. Although CLL has been traditionally considered as a mature B cell leukemia, few independent studies have shown that the genetic alterations may appear in CD34+ hematopoietic progenitors. However, the presenc...
| Autores: | , , , , , , , , , , , , , , , , , , , , , , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2017 |
| País: | España |
| Institución: | Universidad Complutense de Madrid (UCM) |
| Repositorio: | Docta Complutense |
| Idioma: | inglés |
| OAI Identifier: | oai:docta.ucm.es:20.500.14352/93317 |
| Acceso en línea: | https://hdl.handle.net/20.500.14352/93317 |
| Access Level: | acceso abierto |
| Palabra clave: | 616.155.392 Chronic lymphocytic leukemia Next-generation sequencing Hematopoietic progenitors Mutation FISH Chromosomal abnormality Ciencias Biomédicas 24 Ciencias de la Vida |
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Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemiaQuijada-Álamo, MiguelHernández Sánchez, MaríaRobledo, CristinaHernández-Sánchez, Jesús-MaríaBenito, RocíoMontaño, AdriánRodríguez-Vicente, Ana EQuwaider, DaliaMartín, Ana-ÁfricaGarcía-Álvarez, MaríaVidal-Manceñido, María JesúsFerrer-Garrido, GonzaloDelgado-Beltrán, María-PilarGalende, JosefinaRodríguez, Juan-NicolásMartín-Núñez, GuillermoAlonso, José-MaríaGarcía de Coca, AlfonsoQueizán, José A.Sierra, MagdalenaAguilar, CarlosKohlmann, AlexanderHernández,José-ÁngelGonzález, MarcosHernández-Rivas, Jesús-María616.155.392Chronic lymphocytic leukemiaNext-generation sequencingHematopoietic progenitorsMutationFISHChromosomal abnormalityCiencias Biomédicas24 Ciencias de la VidaBackground Chronic lymphocytic leukemia (CLL) is a highly genetically heterogeneous disease. Although CLL has been traditionally considered as a mature B cell leukemia, few independent studies have shown that the genetic alterations may appear in CD34+ hematopoietic progenitors. However, the presence of both chromosomal aberrations and gene mutations in CD34+ cells from the same patients has not been explored. Methods Amplicon-based deep next-generation sequencing (NGS) studies were carried out in magnetically activated-cell-sorting separated CD19+ mature B lymphocytes and CD34+ hematopoietic progenitors (n = 56) to study the mutational status of TP53, NOTCH1, SF3B1, FBXW7, MYD88, and XPO1 genes. In addition, ultra-deep NGS was performed in a subset of seven patients to determine the presence of mutations in flow-sorted CD34+CD19− early hematopoietic progenitors. Fluorescence in situ hybridization (FISH) studies were performed in the CD34+ cells from nine patients of the cohort to examine the presence of cytogenetic abnormalities. Results NGS studies revealed a total of 28 mutations in 24 CLL patients. Interestingly, 15 of them also showed the same mutations in their corresponding whole population of CD34+ progenitors. The majority of NOTCH1 (7/9) and XPO1 (4/4) mutations presented a similar mutational burden in both cell fractions; by contrast, mutations of TP53 (2/2), FBXW7 (2/2), and SF3B1 (3/4) showed lower mutational allele frequencies, or even none, in the CD34+ cells compared with the CD19+ population. Ultra-deep NGS confirmed the presence of FBXW7, MYD88, NOTCH1, and XPO1 mutations in the subpopulation of CD34+CD19− early hematopoietic progenitors (6/7). Furthermore, FISH studies showed the presence of 11q and 13q deletions (2/2 and 3/5, respectively) in CD34+ progenitors but the absence of IGH cytogenetic alterations (0/2) in the CD34+ cells. Combining all the results from NGS and FISH, a model of the appearance and expansion of genetic alterations in CLL was derived, suggesting that most of the genetic events appear on the hematopoietic progenitors, although these mutations could induce the beginning of tumoral cell expansion at different stage of B cell differentiation. Conclusions Our study showed the presence of both gene mutations and chromosomal abnormalities in early hematopoietic progenitor cells from CLL patients.BMCUniversidad Complutense de Madrid20172017-04-1120172017-04-11journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/93317reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)InglésengPI12 00281 Not availablePI15 01471 Not available1172 A 15BIO SA10 141343 A 16RD12 0036 0069open accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/933172026-06-02T12:44:21Z |
| dc.title.none.fl_str_mv |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| title |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| spellingShingle |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia Quijada-Álamo, Miguel 616.155.392 Chronic lymphocytic leukemia Next-generation sequencing Hematopoietic progenitors Mutation FISH Chromosomal abnormality Ciencias Biomédicas 24 Ciencias de la Vida |
| title_short |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| title_full |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| title_fullStr |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| title_full_unstemmed |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| title_sort |
Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia |
| dc.creator.none.fl_str_mv |
Quijada-Álamo, Miguel Hernández Sánchez, María Robledo, Cristina Hernández-Sánchez, Jesús-María Benito, Rocío Montaño, Adrián Rodríguez-Vicente, Ana E Quwaider, Dalia Martín, Ana-África García-Álvarez, María Vidal-Manceñido, María Jesús Ferrer-Garrido, Gonzalo Delgado-Beltrán, María-Pilar Galende, Josefina Rodríguez, Juan-Nicolás Martín-Núñez, Guillermo Alonso, José-María García de Coca, Alfonso Queizán, José A. Sierra, Magdalena Aguilar, Carlos Kohlmann, Alexander Hernández,José-Ángel González, Marcos Hernández-Rivas, Jesús-María |
| author |
Quijada-Álamo, Miguel |
| author_facet |
Quijada-Álamo, Miguel Hernández Sánchez, María Robledo, Cristina Hernández-Sánchez, Jesús-María Benito, Rocío Montaño, Adrián Rodríguez-Vicente, Ana E Quwaider, Dalia Martín, Ana-África García-Álvarez, María Vidal-Manceñido, María Jesús Ferrer-Garrido, Gonzalo Delgado-Beltrán, María-Pilar Galende, Josefina Rodríguez, Juan-Nicolás Martín-Núñez, Guillermo Alonso, José-María García de Coca, Alfonso Queizán, José A. Sierra, Magdalena Aguilar, Carlos Kohlmann, Alexander Hernández,José-Ángel González, Marcos Hernández-Rivas, Jesús-María |
| author_role |
author |
| author2 |
Hernández Sánchez, María Robledo, Cristina Hernández-Sánchez, Jesús-María Benito, Rocío Montaño, Adrián Rodríguez-Vicente, Ana E Quwaider, Dalia Martín, Ana-África García-Álvarez, María Vidal-Manceñido, María Jesús Ferrer-Garrido, Gonzalo Delgado-Beltrán, María-Pilar Galende, Josefina Rodríguez, Juan-Nicolás Martín-Núñez, Guillermo Alonso, José-María García de Coca, Alfonso Queizán, José A. Sierra, Magdalena Aguilar, Carlos Kohlmann, Alexander Hernández,José-Ángel González, Marcos Hernández-Rivas, Jesús-María |
| author2_role |
author author author author author author author author author author author author author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Universidad Complutense de Madrid |
| dc.subject.none.fl_str_mv |
616.155.392 Chronic lymphocytic leukemia Next-generation sequencing Hematopoietic progenitors Mutation FISH Chromosomal abnormality Ciencias Biomédicas 24 Ciencias de la Vida |
| topic |
616.155.392 Chronic lymphocytic leukemia Next-generation sequencing Hematopoietic progenitors Mutation FISH Chromosomal abnormality Ciencias Biomédicas 24 Ciencias de la Vida |
| description |
Background Chronic lymphocytic leukemia (CLL) is a highly genetically heterogeneous disease. Although CLL has been traditionally considered as a mature B cell leukemia, few independent studies have shown that the genetic alterations may appear in CD34+ hematopoietic progenitors. However, the presence of both chromosomal aberrations and gene mutations in CD34+ cells from the same patients has not been explored. Methods Amplicon-based deep next-generation sequencing (NGS) studies were carried out in magnetically activated-cell-sorting separated CD19+ mature B lymphocytes and CD34+ hematopoietic progenitors (n = 56) to study the mutational status of TP53, NOTCH1, SF3B1, FBXW7, MYD88, and XPO1 genes. In addition, ultra-deep NGS was performed in a subset of seven patients to determine the presence of mutations in flow-sorted CD34+CD19− early hematopoietic progenitors. Fluorescence in situ hybridization (FISH) studies were performed in the CD34+ cells from nine patients of the cohort to examine the presence of cytogenetic abnormalities. Results NGS studies revealed a total of 28 mutations in 24 CLL patients. Interestingly, 15 of them also showed the same mutations in their corresponding whole population of CD34+ progenitors. The majority of NOTCH1 (7/9) and XPO1 (4/4) mutations presented a similar mutational burden in both cell fractions; by contrast, mutations of TP53 (2/2), FBXW7 (2/2), and SF3B1 (3/4) showed lower mutational allele frequencies, or even none, in the CD34+ cells compared with the CD19+ population. Ultra-deep NGS confirmed the presence of FBXW7, MYD88, NOTCH1, and XPO1 mutations in the subpopulation of CD34+CD19− early hematopoietic progenitors (6/7). Furthermore, FISH studies showed the presence of 11q and 13q deletions (2/2 and 3/5, respectively) in CD34+ progenitors but the absence of IGH cytogenetic alterations (0/2) in the CD34+ cells. Combining all the results from NGS and FISH, a model of the appearance and expansion of genetic alterations in CLL was derived, suggesting that most of the genetic events appear on the hematopoietic progenitors, although these mutations could induce the beginning of tumoral cell expansion at different stage of B cell differentiation. Conclusions Our study showed the presence of both gene mutations and chromosomal abnormalities in early hematopoietic progenitor cells from CLL patients. |
| publishDate |
2017 |
| dc.date.none.fl_str_mv |
2017 2017-04-11 2017 2017-04-11 |
| dc.type.none.fl_str_mv |
journal article http://purl.org/coar/resource_type/c_6501 VoR http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
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article |
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https://hdl.handle.net/20.500.14352/93317 |
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https://hdl.handle.net/20.500.14352/93317 |
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Inglés eng |
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Inglés |
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eng |
| dc.relation.none.fl_str_mv |
PI12 00281 Not available PI15 01471 Not available 1172 A 15 BIO SA10 14 1343 A 16 RD12 0036 0069 |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ |
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info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ |
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openAccess |
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application/pdf |
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BMC |
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BMC |
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reponame:Docta Complutense instname:Universidad Complutense de Madrid (UCM) |
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Universidad Complutense de Madrid (UCM) |
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