Cellular genomic reporter assays for screening and evaluation of inducers of fetal hemoglobin

Jim Vadolas, Hady Wardan, Michael Orford, Robert Williamson, Panayiotis A. Ioannou

Research output: Contribution to journalReview ArticleResearchpeer-review

24 Citations (Scopus)

Abstract

Reactivation of fetal hemoglobin (HbF) expression using pharmacological agents represents a potential strategy for the therapy of β-thalassemia, sickle cell disease, HbE and other β-hemoglobinopathies. However, the drugs currently available have low efficacy and specificity and are associated with high toxicity. We describe the development of stable cellular genomic reporter assays (GRAs) based on the green fluorescence protein (EGFP) gene under the Gγ-globin promoter in the intact human β-globin locus. We show that human erythroleukemic cell lines stably transfected with a Gγ-EGFP β-globin locus construct can maintain a uniform basal level of EGFP expression over long periods of continuous culture and that induction of EGFP expression parallels the induction of the endogenous globin genes. We compared the EGFP-induction potency of a number of chemotherapeutic agents, including histone deacetylase inhibitors and DNA-binding agents. We show that hydroxyurea and butyrate result in moderate levels of induction (70-80%) but with an additive inductive effect. Among the DNA-binding agents tested, cisplatin was the most potent inducer of HbF expression, (442±32% , a level which is comparable to hemin (764±145%). These results indicate that cellular GRAs containing Gγ-EGFP-modified β-globin locus constructs can be used to develop novel inducers of HbF synthesis for the therapy of β-hemoglobinopathies.

Original languageEnglish
Pages (from-to)223-233
Number of pages11
JournalHuman Molecular Genetics
Volume13
Issue number2
DOIs
Publication statusPublished - 15 Jan 2004
Externally publishedYes

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