siRNA-mediated reduction of alpha-globin results in phenotypic improvements in beta-thalassemic cells.
beta-thalassemia is an inherited hemoglobinopathy caused by defective synthesis of the beta-globin chain of hemoglobin, leading to imbalanced globin chain synthesis. Excess alpha-globin precipitates in erythroid progenitor cells resulting in cell death, ineffective erythropoiesis and severe anemia. Decreased alpha-globin synthesis leads to milder symptoms, exemplified in individuals who co-inherit alpha- and beta-thalassemia. In this study, we investigated the feasibility of utilizing short-interfering RNA (siRNA) to mediate reductions in alpha-globin expression. A number of siRNA sequences targeting murine alpha-globin were tested in hemoglobinized murine erythroleukemic cells. One highly effective siRNA sequence (si-alpha 4) was identified and reduced alpha-globin by approximately 65% at both the RNA and the protein level. Electroporation of si-alpha 4 into murine thalassemic primary erythroid cultures restored alpha :beta-globin ratios to balanced wild-type levels and resulted in detectable phenotypic correction. These results indicate that siRNA-mediated reduction of alpha-globin has potential therapeutic applications in the treatment of beta-thalassemia.
Voon
Hsiao Phin Joanna
HP
Cell and Gene Therapy Research Group, The Murdoch Children's Research Institute, The University of Melbourne, Royal Children's Hospital, Flemington Road, Parkville 3052, Melbourne, Australia.
Wardan
Hady
H
Vadolas
Jim
J
D000818:Animals; D016923:Cell Death; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D005914:Globins / Q000172:deficiency / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000097:blood; D051379:Mice; D018345:Mice, Knockout; D010641:Phenotype; D016133:Polymerase Chain Reaction; D034741:RNA, Small Interfering / Q000235:genetics* / Q000627:therapeutic use; D017086:beta-Thalassemia / Q000097:blood* / Q000235:genetics / Q000473:pathology / Q000628:therapy
D034741:RNA, Small Interfering; D005914:Globins