Optimization of β-globin Stable Expression Using the Third Generation Lentiviral Vector for β-thalassemia Therapy.
10.19746/j.cnki.issn.1009-2137.2022.03.029
- Author:
Zhen YU
1
;
Shuai TONG
1
;
Yue BAI
1
;
Xiao-Song ZHONG
2
Author Information
1. Clinical Gene and Cell Engineering Center, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China.
2. Clinical Gene and Cell Engineering Center, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China,E-mail: zhongxiaosong7113@bjsjth.cn.
- Publication Type:Journal Article
- Keywords:
gene therapy;
lentivirus;
β-globin;
β-thalassemia
- MeSH:
Animals;
Genetic Therapy;
Genetic Vectors;
Humans;
Lentivirus/genetics*;
Leukocytes, Mononuclear;
Mice;
beta-Globins/genetics*;
beta-Thalassemia/therapy*
- From:
Journal of Experimental Hematology
2022;30(3):844-850
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To provide a research basis for a safe and effective cell therapy for β-thalassemia through optimization of HS4 region of the third generation lentiviral vector for stable expression of β-globin.
METHODS:The human β-globin HS4 region in the third generation lentiviral expression vector was optimized to construct the lenti-HBB, and the transcription and translation of β-globin gene were analyzed by RT-PCR and Western blot after the transduction of lenti-HBB in MEL cell line. Furthermore, the erythroid differentiation of CD34+ cells which were transduced lentiviral virus carrying human β-globin from normal human umbilical cord blood cells and peripheral blood cells of patients with β-thalassemia major were confirmed by colony formation assay, cell smear assay and flow cytometry. The safety and effectiveness of the optimized lenti-HBB were verified by NSG mouse in vivo test.
RESULTS:The human β-globin was expressed stably in the MEL cells, and CD34+ cells from health umbilical cord blood as well as PBMC from patient with β-thalassemia major transduced with lenti-HBB could be differentiated to mature red blood cells. The β-globin expression and differentiation in CD34+ cells were demonstrated successfully in the NSG mouse for about 35 months after post-transplant.
CONCLUSION:Stable β-globin expression through the optimization of HS4 from CD34+ in the third generation lentiviral vector is safe and effective for patients with severe β-thalassemia and other β-globin abnormal diseases.
