betaMinor-globin messenger RNA accumulation in reticulocytes governs improved erythropoiesis in beta thalassemic mice after erythropoietin complementary DNA electrotransfer in muscles.
Mechanisms governing the induction of effective erythropoiesis in response to erythropoietin (Epo) oversecretion have been investigated in beta thalassemic C57Bl/6(Hbbth) mice. Naked DNA encoding an expression vector for mouse Epo was introduced into skeletal muscles by electrotransfer. A transient increase of serum Epo concentrations with a proportional augmentation of hematocrit values was observed. Various parameters relevant to beta thalassemia were surveyed in blood samples taken before treatment, at the peak of Epo secretion, and when the phenotype reverted to anemia. We measured globin messenger RNA (mRNA) levels in reticulocytes by real-time quantitative polymerase chain reaction, globin chain synthesis levels, and several indicators of erythrocyte membrane quality, including bound alpha chains, bound immunoglobulins, main protein components, and iron compartmentalization. Data indicated that high serum Epo levels primarily affect betaminor-globin mRNA accumulation in reticulocytes. Other changes subsequent to intense Epo stimulation, like increased betaminor/alpha-globin chain synthesis ratio, reduced levels of alpha chains and immunoglobulins bound to membranes, improved spectrin/band 3 ratio, increased red blood cell survival, and improved erythropoiesis appeared as consequences of increased betaminor-globin mRNA levels. This conclusion is consistent with models postulating that intense Epo stimulation induces the expansion and differentiation of erythroid progenitors committed to fetal erythropoiesis. Although phenotypic correction was partial in mice, and comparable achievements will probably be more difficult to obtain in humans, naked DNA electrotransfer may provide a safe and low-cost method for reassessing the potentials of Epo as an inducer of fetal erythropoiesis reactivation in patients with beta thalassemia.
Samakoglu
S
S
Laboratoire Rétrovirus et Transfert Génétique, CNRS URA 1930; Institut Pasteur, Paris, France.
Fattori
E
E
Lamartina
S
S
Toniatti
C
C
Stockholm
D
D
Heard
J M
JM
Bohl
D
D
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000032:analysis; D002451:Cell Compartmentation; D002454:Cell Differentiation; D018076:DNA, Complementary / Q000008:administration & dosage / Q000235:genetics*; D004195:Disease Models, Animal; D018274:Electroporation; D004912:Erythrocytes / Q000378:metabolism; D015672:Erythroid Precursor Cells / Q000378:metabolism / Q000473:pathology; D004920:Erythropoiesis / Q000235:genetics*; D004921:Erythropoietin / Q000097:blood / Q000235:genetics*; D005816:Genetic Complementation Test; D005914:Globins / Q000235:genetics*; D007273:Injections, Intramuscular; D007501:Iron / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000502:physiology; D012156:Reticulocytes / Q000378:metabolism*; D013049:Spectrin / Q000032:analysis; D014162:Transfection; D017086:beta-Thalassemia / Q000235:genetics*
D001457:Anion Exchange Protein 1, Erythrocyte; D018076:DNA, Complementary; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004921:Erythropoietin; D013049:Spectrin; D005914:Globins; D007501:Iron