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

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.

• Authors: 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
• Journal: PloS one (PLoS One) Vol. 9 Issue 9 Pg. e107006 ( 2014) ISSN: 1932-6203 [Electronic] United States
• PMID: 25225870 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, CD34
• beta-Globins
• gamma-Globins
• Fetal Hemoglobin

Topics

• Animals
• Antigens, CD34 (metabolism)
• Bone Marrow Cells (drug effects, metabolism)
• Chromosomes, Artificial, Yeast
• Drug Discovery
• Drug Evaluation, Preclinical
• Erythroid Precursor Cells (drug effects, metabolism)
• Fetal Hemoglobin (biosynthesis, genetics)
• Gene Expression Regulation (drug effects)
• Gene Targeting
• Genes, Reporter
• Genetic Loci
• Genetic Vectors (genetics)
• Hemoglobinopathies (drug therapy, genetics)
• High-Throughput Screening Assays
• Humans
• Mice
• Mice, Transgenic
• beta-Globins (biosynthesis, genetics)
• gamma-Globins (biosynthesis, genetics)