PMID- 25225870 OWN - NLM STAT- MEDLINE DCOM- 20150527 LR - 20211021 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 9 IP - 9 DP - 2014 TI - A cell-based high-throughput screen for novel chemical inducers of fetal hemoglobin for treatment of hemoglobinopathies. PG - e107006 LID - 10.1371/journal.pone.0107006 [doi] LID - e107006 AB - 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 gamma-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 gamma-globin off-on switch to identify potential lead compounds. Assay systems developed in our labs to understand the mechanisms underlying the gamma- to beta-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 gamma-globin promoter-firefly luciferase beta-globin promoter-Renilla luciferase beta-globin yeast artificial chromosome (gamma-luc beta-luc beta-YAC) transgenic mice, we were able to identify 232 lead chemical compounds that induced gamma-globin 2-fold or higher, with minimal or no beta-globin induction, minimal cytotoxicity and that did not directly influence the luciferase enzyme. Secondary assays in CID-dependent wild-type beta-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. FAU - Peterson, Kenneth R AU - Peterson KR AD - 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. FAU - Costa, Flavia C AU - Costa FC AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Fedosyuk, Halyna AU - Fedosyuk H AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Neades, Renee Y AU - Neades RY AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Chazelle, Allen M AU - Chazelle AM AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Zelenchuk, Lesya AU - Zelenchuk L AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Fonteles, Andrea H AU - Fonteles AH AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Dalal, Parmita AU - Dalal P AD - Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, United States of America. FAU - Roy, Anuradha AU - Roy A AD - High Throughput Screening Laboratory, University of Kansas, Lawrence, Kansas, United States of America. FAU - Chaguturu, Rathnam AU - Chaguturu R AD - High Throughput Screening Laboratory, University of Kansas, Lawrence, Kansas, United States of America. FAU - Li, Biaoru AU - Li B AD - Department of Pediatrics, Georgia Regents University, Augusta, Georgia, United States of America. FAU - Pace, Betty S AU - Pace BS AD - 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. LA - eng GR - HL067336/HL/NHLBI NIH HHS/United States GR - R01 DK081290/DK/NIDDK NIH HHS/United States GR - R56 DK081290/DK/NIDDK NIH HHS/United States GR - R01 HL069234/HL/NHLBI NIH HHS/United States GR - R01 DK061804/DK/NIDDK NIH HHS/United States GR - DK081290/DK/NIDDK NIH HHS/United States GR - P30 GM103495/GM/NIGMS NIH HHS/United States GR - DK061804/DK/NIDDK NIH HHS/United States GR - 5 P30 CA168524/CA/NCI NIH HHS/United States GR - P30 CA168524/CA/NCI NIH HHS/United States GR - R56 HL067336/HL/NHLBI NIH HHS/United States GR - P30 GM103326/GM/NIGMS NIH HHS/United States GR - R01 HL067336/HL/NHLBI NIH HHS/United States GR - 8 P30 GM103495/GM/NIGMS NIH HHS/United States GR - R01 HL111264/HL/NHLBI NIH HHS/United States GR - HL069234/HL/NHLBI NIH HHS/United States GR - P20 RR015563/RR/NCRR NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20140916 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Antigens, CD34) RN - 0 (beta-Globins) RN - 0 (gamma-Globins) RN - 9034-63-3 (Fetal Hemoglobin) SB - IM MH - Animals MH - Antigens, CD34/metabolism MH - Bone Marrow Cells/*drug effects/*metabolism MH - Chromosomes, Artificial, Yeast MH - *Drug Discovery MH - Drug Evaluation, Preclinical MH - Erythroid Precursor Cells/drug effects/metabolism MH - Fetal Hemoglobin/biosynthesis/*genetics MH - Gene Expression Regulation/*drug effects MH - Gene Targeting MH - Genes, Reporter MH - Genetic Loci MH - Genetic Vectors/genetics MH - Hemoglobinopathies/drug therapy/genetics MH - *High-Throughput Screening Assays MH - Humans MH - Mice MH - Mice, Transgenic MH - beta-Globins/biosynthesis/genetics MH - gamma-Globins/biosynthesis/genetics PMC - PMC4165891 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2014/09/17 06:00 MHDA- 2015/05/28 06:00 PMCR- 2014/09/16 CRDT- 2014/09/17 06:00 PHST- 2014/05/06 00:00 [received] PHST- 2014/08/04 00:00 [accepted] PHST- 2014/09/17 06:00 [entrez] PHST- 2014/09/17 06:00 [pubmed] PHST- 2015/05/28 06:00 [medline] PHST- 2014/09/16 00:00 [pmc-release] AID - PONE-D-14-20279 [pii] AID - 10.1371/journal.pone.0107006 [doi] PST - epublish SO - PLoS One. 2014 Sep 16;9(9):e107006. doi: 10.1371/journal.pone.0107006. eCollection 2014.