Abstract
We systematically compared cytokine-mediated increases or decreases in proliferation with globin gene and protein expression in adult human erythroblasts. Despite their opposite effects on growth, stem cell factor (SCF) and transforming growth factor-beta (TGF-B) had synergistic effects with respect to fetal hemoglobin (HbF): average HbF/HbF + adult hemoglobin (HbA) ratio in erythropoietin (EPO) = 1.4 ± 1.0%; EPO + TGF-B = 10.8 ± 1.9%; EPO + SCF = 19.1 ± 6.2%; and EPO + SCF + TGF-B (EST) = 39.3 ± 6.3%. Polymerase chain reaction (PCR) revealed significant increases in gamma-globin transcripts that were balanced by reduced beta-globin transcripts. Single-cell quantitative PCR demonstrated a complete reversal of gamma-globin gene silencing with detectable gamma-globin mRNA in more than 95% of the cells. Immunostaining with HbF antibodies also showed a pancellular distribution in EST (96.2 ± 0.01% HbF positive) compared with a heterocellular distribution in EPO (42.9 ± 0.01% HbF positive). As shown here for the first time, a robust and pancellular reversal of gamma-globin gene silencing among hemoglobinized erythroblasts from adult humans may be achieved in the absence of hereditary mutation or direct genomic manipulation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 387-393 |
| Number of pages | 7 |
| Journal | Blood |
| Volume | 105 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 1 2005 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Immunology
- Biochemistry
- Hematology
- Cell Biology
Cite this
A sustained and pancellular reversal of gamma-globin gene silencing in adult human erythroid precursor cells. / Bhanu, Natarajan V.; Trice, Tiffany A.; Lee, Y. Terry; Gantt, Nicole M.; Oneal, Patricia; Schwartz, Joseph D.; Noel, Pierre; Miller, Jeffery L.
In: Blood, Vol. 105, No. 1, 01.01.2005, p. 387-393.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A sustained and pancellular reversal of gamma-globin gene silencing in adult human erythroid precursor cells
AU - Bhanu, Natarajan V.
AU - Trice, Tiffany A.
AU - Lee, Y. Terry
AU - Gantt, Nicole M.
AU - Oneal, Patricia
AU - Schwartz, Joseph D.
AU - Noel, Pierre
AU - Miller, Jeffery L.
PY - 2005/1/1
Y1 - 2005/1/1
N2 - We systematically compared cytokine-mediated increases or decreases in proliferation with globin gene and protein expression in adult human erythroblasts. Despite their opposite effects on growth, stem cell factor (SCF) and transforming growth factor-beta (TGF-B) had synergistic effects with respect to fetal hemoglobin (HbF): average HbF/HbF + adult hemoglobin (HbA) ratio in erythropoietin (EPO) = 1.4 ± 1.0%; EPO + TGF-B = 10.8 ± 1.9%; EPO + SCF = 19.1 ± 6.2%; and EPO + SCF + TGF-B (EST) = 39.3 ± 6.3%. Polymerase chain reaction (PCR) revealed significant increases in gamma-globin transcripts that were balanced by reduced beta-globin transcripts. Single-cell quantitative PCR demonstrated a complete reversal of gamma-globin gene silencing with detectable gamma-globin mRNA in more than 95% of the cells. Immunostaining with HbF antibodies also showed a pancellular distribution in EST (96.2 ± 0.01% HbF positive) compared with a heterocellular distribution in EPO (42.9 ± 0.01% HbF positive). As shown here for the first time, a robust and pancellular reversal of gamma-globin gene silencing among hemoglobinized erythroblasts from adult humans may be achieved in the absence of hereditary mutation or direct genomic manipulation.
AB - We systematically compared cytokine-mediated increases or decreases in proliferation with globin gene and protein expression in adult human erythroblasts. Despite their opposite effects on growth, stem cell factor (SCF) and transforming growth factor-beta (TGF-B) had synergistic effects with respect to fetal hemoglobin (HbF): average HbF/HbF + adult hemoglobin (HbA) ratio in erythropoietin (EPO) = 1.4 ± 1.0%; EPO + TGF-B = 10.8 ± 1.9%; EPO + SCF = 19.1 ± 6.2%; and EPO + SCF + TGF-B (EST) = 39.3 ± 6.3%. Polymerase chain reaction (PCR) revealed significant increases in gamma-globin transcripts that were balanced by reduced beta-globin transcripts. Single-cell quantitative PCR demonstrated a complete reversal of gamma-globin gene silencing with detectable gamma-globin mRNA in more than 95% of the cells. Immunostaining with HbF antibodies also showed a pancellular distribution in EST (96.2 ± 0.01% HbF positive) compared with a heterocellular distribution in EPO (42.9 ± 0.01% HbF positive). As shown here for the first time, a robust and pancellular reversal of gamma-globin gene silencing among hemoglobinized erythroblasts from adult humans may be achieved in the absence of hereditary mutation or direct genomic manipulation.
UR - http://www.scopus.com/inward/record.url?scp=11144220892&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=11144220892&partnerID=8YFLogxK
U2 - 10.1182/blood-2004-04-1599
DO - 10.1182/blood-2004-04-1599
M3 - Article
C2 - 15367428
AN - SCOPUS:11144220892
VL - 105
SP - 387
EP - 393
JO - Blood
JF - Blood
SN - 0006-4971
IS - 1
ER -