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Deficiency of ribosomal protein S19 in CD34+ cells generated by siRNA blocks erythroid development and mimics defects seen in Diamond-Blackfan anemia.

Flygare, Johan LU ; Kiefer, Thomas LU ; Miyake, Koichi LU ; Utsugisawa, Taiju LU ; Hamaguchi, Isao LU ; Da Costa, Lydie; Richter, Johan LU ; Davey, Edward J; Matsson, Hans and Dahl, Niklas, et al. (2005) In Blood 105(12). p.4627-4634
Abstract
Diamond-Blackfan anemia (DBA) is a congenital red cell aplasia in which 25% of the patients have a mutation in the ribosomal protein S19 (RPS19) gene. To study effects of RPS19 deficiency in hematopoiesis we transduced CD34(+) umbilical cord blood (CB) and bone marrow (BM) cells with 3 lentiviral vectors expressing small interfering RNA (siRNA) against RPS19 and 1 scrambled control vector. All vectors also express green fluorescent protein (GFP). Transduction with the siRNA vectors reduced RPS19 mRNA levels to various degrees, which resulted in erythroid defects, correlating to the degree of RPS19 down-regulation, and was rescued by expression of an siRNA-resistant RPS19 transcript. Erythroid colony formation capacity conjointly decreased... (More)
Diamond-Blackfan anemia (DBA) is a congenital red cell aplasia in which 25% of the patients have a mutation in the ribosomal protein S19 (RPS19) gene. To study effects of RPS19 deficiency in hematopoiesis we transduced CD34(+) umbilical cord blood (CB) and bone marrow (BM) cells with 3 lentiviral vectors expressing small interfering RNA (siRNA) against RPS19 and 1 scrambled control vector. All vectors also express green fluorescent protein (GFP). Transduction with the siRNA vectors reduced RPS19 mRNA levels to various degrees, which resulted in erythroid defects, correlating to the degree of RPS19 down-regulation, and was rescued by expression of an siRNA-resistant RPS19 transcript. Erythroid colony formation capacity conjointly decreased with RPS19 levels in CD34(+) CB and BM cells. In liquid culture supporting erythroid differentiation, RPS19-slienced as well as DBA patient CD34(+) cells exhibited reduced proliferative capacity and impaired erythroid differentiation resulting in fewer erythroid colony-forming units (CFU-Es). When assaying myeloid development, a less pronounced influence on proliferation was seen. This study shows for the first time that RPS19 silencing decreases the proliferative capacity of hematopoietic progenitors and leads to a defect in erythroid development. (Less)
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Contribution to journal
publication status
published
subject
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Blood
volume
105
issue
12
pages
4627 - 4634
publisher
American Society of Hematology
external identifiers
  • pmid:15626736
  • wos:000229757300023
  • scopus:20444363463
ISSN
1528-0020
DOI
10.1182/blood-2004-08-3115
language
English
LU publication?
yes
id
f3554122-1823-4af5-b452-de313036650c (old id 133365)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15626736&dopt=Abstract
date added to LUP
2007-07-11 10:58:52
date last changed
2017-07-30 03:34:39
@article{f3554122-1823-4af5-b452-de313036650c,
  abstract     = {Diamond-Blackfan anemia (DBA) is a congenital red cell aplasia in which 25% of the patients have a mutation in the ribosomal protein S19 (RPS19) gene. To study effects of RPS19 deficiency in hematopoiesis we transduced CD34(+) umbilical cord blood (CB) and bone marrow (BM) cells with 3 lentiviral vectors expressing small interfering RNA (siRNA) against RPS19 and 1 scrambled control vector. All vectors also express green fluorescent protein (GFP). Transduction with the siRNA vectors reduced RPS19 mRNA levels to various degrees, which resulted in erythroid defects, correlating to the degree of RPS19 down-regulation, and was rescued by expression of an siRNA-resistant RPS19 transcript. Erythroid colony formation capacity conjointly decreased with RPS19 levels in CD34(+) CB and BM cells. In liquid culture supporting erythroid differentiation, RPS19-slienced as well as DBA patient CD34(+) cells exhibited reduced proliferative capacity and impaired erythroid differentiation resulting in fewer erythroid colony-forming units (CFU-Es). When assaying myeloid development, a less pronounced influence on proliferation was seen. This study shows for the first time that RPS19 silencing decreases the proliferative capacity of hematopoietic progenitors and leads to a defect in erythroid development.},
  author       = {Flygare, Johan and Kiefer, Thomas and Miyake, Koichi and Utsugisawa, Taiju and Hamaguchi, Isao and Da Costa, Lydie and Richter, Johan and Davey, Edward J and Matsson, Hans and Dahl, Niklas and Wiznerowicz, Maciej and Trono, Didier and Karlsson, Stefan},
  issn         = {1528-0020},
  language     = {eng},
  number       = {12},
  pages        = {4627--4634},
  publisher    = {American Society of Hematology},
  series       = {Blood},
  title        = {Deficiency of ribosomal protein S19 in CD34+ cells generated by siRNA blocks erythroid development and mimics defects seen in Diamond-Blackfan anemia.},
  url          = {http://dx.doi.org/10.1182/blood-2004-08-3115},
  volume       = {105},
  year         = {2005},
}