A novel transgenic mouse model produced from lentiviral germline integration for the study of beta-thalassemia gene therapy.
BACKGROUND
beta-thalassemia is one of the most common genetic diseases in the world and requires extensive therapy. Lentiviral-mediated gene therapy has been successfully exploited in the treatment of beta-thalassemia and showed promise in clinical application. Using a human beta-globin transgenic mouse line in a beta-thalassemia diseased model generated with a lentiviral-mediated approach, we investigate the stable therapeutic effect on a common thalassemia syndrome.


METHODS
Human beta-globin gene lentiviral vector was constr ucted, followed by subzonal microinjection into single-cell embryos of beta(IVS-2-654)-thalassemia mice to generate a transgenic line. Human beta-globin gene expression was examined with RT-PCR, Western-blotting and ELISA. The hematologic parameters and tissue pathology were investigated over time in founder mice and their off-spring.


RESULTS
Transgenic mice with stable expression of the lentivirus carrying human beta-globin gene were obtained. A marked improvement in red blood cell indices and a dramatic reduction in red blood cell anisocytosis, poikilocytosis and target cells were observed. Nucleated cell proportion was greatly decreased in bone marrow, and splenomegaly with extramedullary hematopoiesis was ameliorated. Iron deposition in liver was also reduced. There was a two-fold increase in the survival rate of the beta(IVS-2-654) mice carrying human beta-globin transgene. Significantly, the germline integration of the lentiviral construct was obtained and stable hematologic phenotype correction was observed over the next two generations of the transgenic mice.


CONCLUSIONS
The generation of human beta-globin transgenic mice in a beta(IVS-2-654)-thalassemia mouse mediated with lentiviral vectors provides a useful model and offers an attractive means to investigate the transgenic stable therapeutic effect in beta-thalassemia.
Li
Wei
W
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiaotong University School of Medicine, 1400/24 West Beijing Road, Shanghai, China.
Xie
Shuyang
S
Guo
Xinbing
X
Gong
Xiuli
X
Wang
Shu
S
Lin
Dan
D
Zhang
Jingzhi
J
Ren
Zhaorui
Z
Huang
Shuzhen
S
Zeng
Fanyi
F
Zeng
Yitao
Y
D000818:Animals; D001853:Bone Marrow / Q000473:pathology; D004195:Disease Models, Animal; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use*; D018095:Germ-Line Mutation; D005914:Globins / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008099:Liver / Q000473:pathology; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic / Q000235:genetics*; D008845:Microinjections; D016254:Mutagenesis, Insertional; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D013154:Spleen / Q000473:pathology; D016662:Virus Integration; D017086:beta-Thalassemia / Q000235:genetics* / Q000473:pathology / Q000628:therapy
D011993:Recombinant Fusion Proteins; D005914:Globins