[Optimization of β-globin Stable Expression Using the Third Generation Lentiviral Vector for β-thalassemia Therapy].

Area of Science:

• Gene Therapy
• Hematology
• Molecular Biology

Background:

• Beta-thalassemia is a severe inherited blood disorder caused by mutations in the beta-globin gene.
• Current treatments are limited and often involve lifelong transfusions or bone marrow transplantation.
• Lentiviral vectors offer a promising platform for gene therapy, but require optimization for stable gene expression.

Purpose of the Study:

• To optimize the HS4 region of a third-generation lentiviral vector for stable beta-globin expression.
• To establish a safe and effective cell therapy for beta-thalassemia.

Main Methods:

• Optimization of the HS4 region in a lentiviral vector (lenti-HBB).
• Analysis of beta-globin transcription and translation in MEL cells via RT-PCR and Western blot.
• Assessment of erythroid differentiation in CD34+ cells from healthy donors and beta-thalassemia patients using colony formation, cell smear, and flow cytometry assays.
• In vivo safety and efficacy evaluation in NSG mice.

Main Results:

• Stable beta-globin expression was achieved in MEL cells.
• Transduced CD34+ cells from healthy donors and beta-thalassemia patients differentiated into mature red blood cells.
• Successful in vivo engraftment and erythroid differentiation in NSG mice for up to 3-5 months post-transplant.

Conclusions:

• Optimization of the HS4 region in lentiviral vectors enables stable beta-globin expression.
• This approach is safe and effective for treating severe beta-thalassemia and other beta-globinopathies.
• The study provides a strong research basis for developing lentiviral vector-based cell therapies.