Development and characterization of K562 cell clones expressing BCL11A-XL: Decreased hemoglobin production with fetal hemoglobin inducers and its rescue with mithramycin.
Induction of fetal hemoglobin (HbF) is considered a promising strategy in the treatment of β-thalassemia, in which production of adult hemoglobin (HbA) is impaired by mutations affecting the β-globin gene. Recent results indicate that B-cell lymphoma/leukemia 11A (BCL11A) is a major repressor of γ-globin gene expression. Therefore, disrupting the binding of the BCL11A transcriptional repressor complex to the γ-globin gene promoter provides a novel approach for inducing expression of the γ-globin genes. To develop a cellular screening system for the identification of BCL11A inhibitors, we produced K562 cell clones with integrated copies of a BCL11A-XL expressing vector. We characterized 12 K562 clones expressing different levels of BCL11A-XL and found that a clear inverse relationship does exist between the levels of BCL11A-XL and the extent of hemoglobinization induced by a panel of HbF inducers. Using mithramycin as an inducer, we found that this molecule was the only HbF inducer efficient in rescuing the ability to differentiate along the erythroid program, even in K562 cell clones expressing high levels of BCL11A-XL, suggesting that BCL11A-XL activity is counteracted by mithramycin.
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy; Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Alessia
A
Finotti
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Jessica
J
Gasparello
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy; Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Giulia
G
Breveglieri
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy; Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Lucia Carmela
LC
Cosenza
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Giulia
G
Montagner
Department of biology, Pomezia, Italy.
Alberto
A
Bresciani
Department of biology, Pomezia, Italy.
Sergio
S
Altamura
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Nicoletta
N
Bianchi
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Elisa
E
Martini
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Eleonora
E
Gallerani
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy; Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Monica
M
Borgatti
Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy; Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy. Electronic address: gam@unife.it.
Roberto
R
Gambari
D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D005319:Fetal Hemoglobin / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects*; D005822:Genetic Vectors; D006801:Humans; D020014:K562 Cells; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D008926:Plicamycin / Q000494:pharmacology*; D012097:Repressor Proteins
C486563:BCL11A protein, human; D002352:Carrier Proteins; D009687:Nuclear Proteins; D012097:Repressor Proteins; D005319:Fetal Hemoglobin; D008926:Plicamycin