title: Hydroxyurea responses and fetal hemoglobin induction in beta-thalassemia/HbE patients' peripheral blood erythroid cell culture.
abstract: Due to genetic heterogeneity of beta-thalassemia (beta-thal) patients, several efforts have been undertaken to determine the efficacy of hydroxyurea treatment. The aim of this work is to determine the responder and nonresponder for hydroxyurea treatment in beta-thal intermedia based on gamma-globin mRNA and fetal hemoglobin (HbF) induction in human erythroid progenitor cells purified from a patient's peripheral blood. Eighteen beta-thal/hemoglobin E patients [13 beta(E)/codon41/42(-TCTT), 4 beta(E)/codon17, and 1 beta(E)/IVS-654], requiring blood transfusion occasionally, with Hb levels of 5.20-8.50 g/dl were studied. The relative levels of gamma-globin mRNA was measured by real-time reverse-transcription polymerase chain reaction and HbF by high-performance liquid chromatography. The results indicated that erythroid progenitor cells treated with 30 mumol/l hydroxyurea for 96 h preferentially enhanced (G)gamma-and (A)gamma-globin mRNA. The mean values of (G)gamma-globin mRNA fold induction were higher than (A)gamma-globin mRNA (12+/-4 vs 4+/-0.30), the Pearson's correlation of (G)gamma-and (A)gamma-globin mRNA was r=0.80. Induction of (G)gamma/(A)gamma globin mRNA is up to ninefold. A 30% increase in the proportion of HbF out of the total Hb was found in cultures derived from four patients, 20-30% in cultures from nine patients, and less than 20% in cultures from five patients. In cultures from only two patients, increase in the proportion of HbF was less than 3%, and (G)gamma/(A)gamma globin mRNA is less than 0.50.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand. ywwatana@yahoo.com
authors.forename: Ramida
authors.initials: R
authors.lastname: Watanapokasin
authors.forename: Duangmanee
authors.initials: D
authors.lastname: Sanmund
authors.forename: Pranee
authors.initials: P
authors.lastname: Winichagoon
authors.forename: Koichiro
authors.initials: K
authors.lastname: Muta
authors.forename: Suthat
authors.initials: S
authors.lastname: Fucharoen
mesh_terms: D001803:Blood Transfusion; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003062:Codon / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015672:Erythroid Precursor Cells / Q000187:drug effects / Q000378:metabolism*; D004920:Erythropoiesis / Q000187:drug effects*; D005260:Female; D005319:Fetal Hemoglobin / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology*; D008297:Male; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015854:Up-Regulation / Q000187:drug effects; D017086:beta-Thalassemia / Q000235:genetics / Q000378:metabolism* / Q000628:therapy
chemical_list: D003062:Codon; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D005319:Fetal Hemoglobin; D006918:Hydroxyurea