bT87Q-globin gene therapy reduces sickle hemoglobin production, allowing for ex vivo anti-sickling activity in human erythroid cells

We performed RNA-seq analysis on immortalized human erythroid cell line (HUDEP-2), that had been engineered to carry the sickle cell disease (SCD) mutation (sHUDEP-2), and on healthy donor mobilized CD34+ derived cells after transduction with a control GFP lentivirus and lentivirus expressing ßT87Q-globin. Our results show activation of inflammation- and proliferation-related programs, suggesting minimal changes of background gene expression except for ßT87Q-globin expression and endogenous ß/ßs-globin suppression. Overall design: RNA-seq was performed on non-transduced CD34+ derived human primary cells and on CD34+ derived human primary cells transduced with a control GFP lenitviral vector or ßT87Q-globin lentiviral vector at the CD71high/CD235ahigh stage of erythroid development (day 9 of culture) and on non-transduced sHUDEP-2 cells and on sHUDEP-2 cells transduced with a control GFP or ßT87Q-globin lentiviral vector at the CD71high/CD235ahigh stage (day 6 of culture).