A cell-based high-throughput screen for novel chemical inducers of fetal hemoglobin for treatment of hemoglobinopathies

Kenneth R. Peterson, Flávia C. Costa, Halyna Fedosyuk, Renee Y. Neades, Allen M. Chazelle, Lesya Zelenchuk, Andrea H. Fonteles, Parmita Dalal, Anuradha Roy, Rathnam Chaguturu, Biaoru Li, Betty S. Pace

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Decades of research have established that the most effective treatment for sickle cell disease (SCD) is increased fetal hemoglobin (HbF). Identification of a drug specific for inducing γ-globin expression in pediatric and adult patients, with minimal off-target effects, continues to be an elusive goal. One hurdle has been an assay amenable to a high-throughput screen (HTS) of chemicals that displays a robust γ-globin off-on switch to identify potential lead compounds. Assay systems developed in our labs to understand the mechanisms underlying the γ- to β-globin gene expression switch during development has allowed us to generate a cell-based assay that was adapted for a HTS of 121,035 compounds. Using chemical inducer of dimerization (CID)-dependent bone marrow cells (BMCs) derived from human γ-globin promoter-firefly luciferase β-globin promoter-Renilla luciferase β-globin yeast artificial chromosome (γ-luc β-luc β-YAC) transgenic mice, we were able to identify 232 lead chemical compounds that induced γ-globin 2-fold or higher, with minimal or no β-globin induction, minimal cytotoxicity and that did not directly influence the luciferase enzyme. Secondary assays in CID dependent wild-type β-YAC BMCs and human primary erythroid progenitor cells confirmed the induction profiles of seven of the 232 hits that were cherry-picked for further analysis.

Original languageEnglish (US)
Article numbere107006
JournalPloS one
Volume9
Issue number9
DOIs
StatePublished - Sep 16 2014

Fingerprint

Fetal Hemoglobin
Hemoglobinopathies
Globins
hemoglobin
luciferase
Throughput
dimerization
bone marrow cells
assays
Assays
yeast artificial chromosomes
promoter regions
cells
sickle cell anemia
Dimerization
chemical compounds
Therapeutics
Bone Marrow Cells
stem cells
cytotoxicity

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

A cell-based high-throughput screen for novel chemical inducers of fetal hemoglobin for treatment of hemoglobinopathies. / Peterson, Kenneth R.; Costa, Flávia C.; Fedosyuk, Halyna; Neades, Renee Y.; Chazelle, Allen M.; Zelenchuk, Lesya; Fonteles, Andrea H.; Dalal, Parmita; Roy, Anuradha; Chaguturu, Rathnam; Li, Biaoru; Pace, Betty S.

In: PloS one, Vol. 9, No. 9, e107006, 16.09.2014.

Research output: Contribution to journalArticle

Peterson, KR, Costa, FC, Fedosyuk, H, Neades, RY, Chazelle, AM, Zelenchuk, L, Fonteles, AH, Dalal, P, Roy, A, Chaguturu, R, Li, B & Pace, BS 2014, 'A cell-based high-throughput screen for novel chemical inducers of fetal hemoglobin for treatment of hemoglobinopathies', PloS one, vol. 9, no. 9, e107006. https://doi.org/10.1371/journal.pone.0107006
Peterson, Kenneth R. ; Costa, Flávia C. ; Fedosyuk, Halyna ; Neades, Renee Y. ; Chazelle, Allen M. ; Zelenchuk, Lesya ; Fonteles, Andrea H. ; Dalal, Parmita ; Roy, Anuradha ; Chaguturu, Rathnam ; Li, Biaoru ; Pace, Betty S. / A cell-based high-throughput screen for novel chemical inducers of fetal hemoglobin for treatment of hemoglobinopathies. In: PloS one. 2014 ; Vol. 9, No. 9.
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