RT Journal Article
T1 Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia
A1 Quijada-Álamo, Miguel
A1 Hernández Sánchez, María
A1 Robledo, Cristina
A1 Hernández-Sánchez, Jesús-María
A1 Benito, Rocío
A1 Montaño, Adrián
A1 Rodríguez-Vicente, Ana E
A1 Quwaider, Dalia
A1 Martín, Ana-África
A1 García-Álvarez, María
A1 Vidal-Manceñido, María Jesús
A1 Ferrer-Garrido, Gonzalo
A1 Delgado-Beltrán, María-Pilar
A1 Galende, Josefina
A1 Rodríguez, Juan-Nicolás
A1 Martín-Núñez, Guillermo
A1 Alonso, José-María
A1 García de Coca, Alfonso
A1 Queizán, José A.
A1 Sierra, Magdalena
A1 Aguilar, Carlos
A1 Kohlmann, Alexander
A1 Hernández, José-Ángel
A1 González, Marcos
A1 Hernández-Rivas, Jesús-María
AB BackgroundChronic 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.MethodsAmplicon-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.ResultsNGS 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.ConclusionsOur study showed the presence of both gene mutations and chromosomal abnormalities in early hematopoietic progenitor cells from CLL patients.
PB BMC
YR 2017
FD 2017-04-11
LK https://hdl.handle.net/20.500.14352/93317
UL https://hdl.handle.net/20.500.14352/93317
LA eng
NO Quijada-Álamo M, Hernández-Sánchez M, Robledo C, Hernández-Sánchez J-M, Benito R, Montaño A, et al. Next-generation sequencing and FISH studies reveal the appearance of gene mutations and chromosomal abnormalities in hematopoietic progenitors in chronic lymphocytic leukemia. J Hematol Oncol 2017;10:83. https://doi.org/10.1186/s13045-017-0450-y.
NO Fondo de Investigaciones Sanitarias
NO Instituto de Salud Carlos III
NO European Regional Development Fund(ERDF) “Una manera de hacer Europa"
NO Consejería de Educación, Junta de Castilla y León
NO Gerencia Regional de Salud de Castilla y León
NO Fundación Española de Hematología y Hemoterapia
NO Red Temática de Investigación Cooperativa en Cáncer
NO Ministerio de Economía y Competitividad (España)
NO Fundación “Memoria Don Samuel Solórzano Barruso” 2016
NO European Union Seventh Framework Programme
NO Fondo Social Europeo
DS Docta Complutense
RD 5 abr 2025