Gene transfer. A potential approach to gene therapy for sickle cell disease

The current status of gene transfer experiments indicates that it is possible to provide (1) safe and efficient retroviral packaging lines for gene transfer; and (2) vectors containing the human beta-globin genes and selectable marker genes which can be transmitted into erythroid cells and are appropriately expressed. In animal autologous bone marrow transplantation experiments, stable and high-level expression of retroviral vectors containing human beta-globin genes has not yet been achieved. New retroviral vectors are being tested that contain different components of the retrovirus as well as the newly described enhancer elements 5' to the epsilon gene and surrounding the beta-globin gene. The long-term goal of human gene therapy for sickle cell disease then consists of constructing optimally safe and efficient retroviral packaging lines as well as retroviral vectors containing the human beta-globin gene and selectable markers such as the neoR gene. One would then remove bone marrow cells from patients with sickle cell disease, transfer the retroviral vectors into the bone marrow cells, and subject the cells to G418 selection in vitro. Next, one would ablate the host bone marrow and autotransplant the manipulated bone marrow bearing the retroviral vector. Finally, one would analyze the reconstituted bone marrow for human beta-globin gene expression. These experiments must await the demonstration of safe and efficient gene transfer in animals, and particularly experiments must be done in monkeys prior to the use of these approaches in humans. Alternative approaches to gene therapy include direct correction of the defect in the beta-globin gene by site-specific recombination of the defective gene with incoming normal gene sequences. This technology is not yet achievable, although preliminary experiments have been performed in which gene correction at a low frequency has been obtained. The obvious advantage of this approach is that the native human beta gene would be reconstituted. Site-specific recombination and addition of normal beta genes to the human genome represent exciting and feasible approaches to human gene therapy using the extraordinary resources of modern molecular and cellular biology. Success in treating disorders of human hemoglobin only awaits additional technical advances for increasing the efficiency of gene transfer and the level of gene expression.