22.7 Applications

Sleeping beauty and piggyBac transposon system have helped in correction of various diseases.

Table 22.2 List of diseases corrected with sleeping beauty and piggyBac :

(i) Genetic modifications of human T lymphocytes

For immunotherapeutic purposes peripheral blood and umbilical cord T cells have been successfully modified with both viral and non-viral gene delivery. This method has been used both for the treatment of viral infections and Epstein Barr virus (EBV) associated lymphoma post autologous bone marrow transplantation. The generation of clinical grade T cells with the use of viral vectors is expensive, time intensive and not free of risks. Modification of peripheral blood mononuclear cells with a CD19-specific chimeric antigen receptor (CAR) has been successfully done with the help of sleeping beauty system. Generation of CAR+ T cells from modified PBMCs help in keeping intact CD4+, CD8+, central memory and effector-effector cell phenotypes. Stable transgene expressions in human T lymphocytes have been standardized in piggyBac system. Multiple transgenes have been used to modify primary lymphocytes in order to redirect their specificity for CD19 which makes them resistant to off target the effects of chemothererapeutic drugs like rapamycin. Human epidermal growth factor receptor-2 specific CAR (HER2-CAR) have been successfully modified with cytotoxic T lymphocytes specific for Epstein Barr Virus (EBV). Food and Drug Administration (FDA) has approved the first clinical trial which involves transposon modified autologous T cells with a second generation CD19-specific CAR.