UCLA

This work encompasses the development of lentiviral technology, and the assessment of viral vector-mediated gene therapy approaches for genetic disorders. Chapter 1 illustrates the development of a high-titer bifunctional lentiviral vector in a vector backbone that has reduced size, high vector yields, and efficient gene transfer to human CD34+ hematopoietic stem and progenitor cells for sickle cell disease (SCD). This lentiviral vector induces high levels of anti-sickling hemoglobins, while concurrently reducing sickle hemoglobin in transduced SCD patient CD34+ cells differentiated into erythrocytes and in the SCD Berkeley mouse model in vivo. Chapter 2 highlights DNA sequencing of peripheral blood cells from Adenosine Deaminase Severe Combined Immunodeficiency (ADA-SCID) patients treated with autologous CD34+ cells transduced with either a γ-retroviral or lentiviral ADA gene vector to assess transgene mutational profiles. We observed reverse transcriptase mutations as well as APOBEC3 guanine (G) to adenosine (A) mutational signatures.