No citation available. Published: 2005.06.14
Benjamin L. Ebert, Michele M. Lee, Jennifer L. Pretz, Aravind Subramanian, Raymond Mak, Todd R. Golub, Colin A. Sieff
Diamond Blackfan Anemia (DBA), a congenital erythroblastopenia, is a model disease for the study of erythroid differentiation, but is poorly understood. RPS19 is the only gene yet to have been associated with DBA, but its relevance to erythroid differentiation is unclear. The molecular basis for the stimulation of erythropoiesis by glucocorticoids in patients with DBA has not been identified. We demonstrate that targeted degradation of the RPS19 gene, through retroviral expression of short hairpin RNAs (shRNAs), in cultured human CD34+ cells blocks the proliferation and differentiation of erythroid progenitor cells. Treatment of RPS19 deficient cells with dexamethasone restores erythroid differentiation to normal levels. We investigated the molecular basis of pharmacologic therapies for DBA using oligonucleotide microarrays to survey gene expression in CD34+ cells treated with combinations of dexamethasone, erythropoietin, stem cell factor, and interleukin-3. Dexamethasone did not alter expression of RPS19, but activated a genetic program that includes a set of key hematopoietic regulatory genes. Genes specific to erythroid progenitor cells were up-regulated by dexamethasone, while genes specific to non-erythroid lineages were down-regulated. Deficiency of RPS19 therefore blocks proliferation of immature erythroid progenitor cells, and dexamethasone activates proliferation of the same cell population through mechanisms independent of RPS19.
| Description | Link/Filename |
|---|---|
| Manuscript | Ebert, RPS19 and dex in DBA, Blood 2005.pdf |
| Commentary in Blood | RPS19 paper commentary, Blood 2005.pdf |
| Gene expression data: CEL files | DBA_cel_files.zip |
| Supplementary Figure S1 | Figure S1.pdf |
| Supplementary Table S1 | TableS1.xls |
| Supplementary Table S2 | TableS2.xls |
| Supplementary Table S3 | TableS3.xls |
| Supplementary Table S4 | TableS4.xls |
| Supplementary Table S5 | TableS5.xls |