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

Autor:	Peterson KR; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America; Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Costa FC; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Fedosyuk H; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Neades RY; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Chazelle AM; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Zelenchuk L; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Fonteles AH; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Dalal P; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America., Roy A; High Throughput Screening Laboratory, University of Kansas, Lawrence, Kansas, United States of America., Chaguturu R; High Throughput Screening Laboratory, University of Kansas, Lawrence, Kansas, United States of America., Li B; Department of Pediatrics, Georgia Regents University, Augusta, Georgia, United States of America., Pace BS; Department of Pediatrics, Georgia Regents University, Augusta, Georgia, United States of America; Department of Molecular and Cell Biology, Georgia Regents University, Augusta, Georgia, United States of America.
Jazyk:	angličtina
Zdroj:	PloS one [PLoS One] 2014 Sep 16; Vol. 9 (9), pp. e107006. Date of Electronic Publication: 2014 Sep 16 (Print Publication: 2014).
DOI:	10.1371/journal.pone.0107006
Abstrakt:	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.
Databáze:	MEDLINE
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