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 ?-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 ?-globin off-on switch to identify potential lead compounds. Assay systems developed in our labs to understand the mechanisms underlying the ?- to ?-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 ?-globin promoter-firefly luciferase ?-globin promoter-Renilla luciferase ?-globin yeast artificial chromosome (?-luc ?-luc ?-YAC) transgenic mice, we were able to identify 232 lead chemical compounds that induced ?-globin 2-fold or higher, with minimal or no ?-globin induction, minimal cytotoxicity and that did not directly influence the luciferase enzyme. Secondary assays in CID-dependent wild-type ?-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.

SUBMITTER: Peterson KR 

PROVIDER: S-EPMC4165891 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature