Production of beta-globin and adult hemoglobin following G418 treatment of erythroid precursor cells from homozygous beta(0)39 thalassemia patients.
In several types of thalassemia (including beta(0)39-thalassemia), stop codon mutations lead to premature translation termination and to mRNA destabilization through nonsense-mediated decay. Drugs (for instance aminoglycosides) can be designed to suppress premature termination, inducing a ribosomal readthrough. These findings have introduced new hopes for the development of a pharmacologic approach to the cure of this disease. However, the effects of aminoglycosides on globin mRNA carrying beta-thalassemia stop mutations have not yet been investigated. In this study, we have used a lentiviral construct containing the beta(0)39-thalassemia globin gene under control of the beta-globin promoter and a LCR cassette. We demonstrated by fluorescence-activated cell sorting (FACS) analysis the production of beta-globin by K562 cell clones expressing the beta(0)39-thalassemia globin gene and treated with G418. More importantly, after FACS and high-performance liquid chromatography (HPLC) analyses, erythroid precursor cells from beta(0)39-thalassemia patients were demonstrated to be able to produce beta-globin and adult hemoglobin after treatment with G418. This study strongly suggests that ribosomal readthrough should be considered a strategy for developing experimental strategies for the treatment of beta(0)-thalassemia caused by stop codon mutations. Am. J. Hematol., 2009. (c) 2009 Wiley-Liss, Inc.
Salvatori
Francesca
F
BioPharmaNet, Department of Biochemistry and Molecular Biology, Ferrara University, Ferrara, Italy.
Breveglieri
Giulia
G
Zuccato
Cristina
C
Finotti
Alessia
A
Bianchi
Nicoletta
N
Borgatti
Monica
M
Feriotto
Giordana
G
Destro
Federica
F
Canella
Alessandro
A
Brognara
Eleonora
E
Lampronti
Ilaria
I
Breda
Laura
L
Rivella
Stefano
S
Gambari
Roberto
R
D002478:Cells, Cultured; D018389:Codon, Nonsense; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000378:metabolism*; D005839:Gentamicins / Q000494:pharmacology*; D006454:Hemoglobins / Q000096:biosynthesis* / Q000235:genetics; D006720:Homozygote; D006801:Humans; D020014:K562 Cells; D055544:beta-Globins / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D017086:beta-Thalassemia / Q000097:blood / Q000188:drug therapy* / Q000235:genetics
D018389:Codon, Nonsense; D005839:Gentamicins; D006454:Hemoglobins; D055544:beta-Globins; C010680:antibiotic G 418