Preclinical safety assessment of modified gamma globin lentiviral vector-mediated autologous hematopoietic stem cell gene therapy for hemoglobinopathies

by Mohammad Shadid, Archana Shrestha, Punam Malik

Previously, we reported the development of a human ^Aγ-globin gene lentivirus (LV), G^bG, which expresses high levels of HbF to correct the sickle cell anemia (SCA) phenotype in the Berkeley SCA mouse model, and then modified the γ-globin gene by substituting glycine at codon 16 with aspartic acid in the ^Aγ-globin gene to generate G^bG^M LV. In the present study, we evaluated the long-term safety of human ^Aγ-globin gene carrying G^bG^M LV in wild-type mice after primary and secondary transplants of G^bG^M-modified hematopoietic stem cells (HSC) over 18 months. The safety of the G^bG^M bone marrow transplant was assessed by monitoring the effects on body weight, hematology, histopathology, malignancy formation, and survival. Mice transplanted with Mock-transduced and spleen focus forming virus (SFFV) γ-retroviral vector (RV)-transduced HSC served as negative and positive controls, respectively. The mean donor-cell engraftment was comparable across Mock, G^bG^M LV, and SFFV RV groups. There were no significant differences in body weight, clinical signs, immunophenotype, or histopathology in the G^bG^M-treated mice compared to controls. Four SFFV RV-treated mice, but none of the G^bG^M-treated mice, developed donor-derived, vector-positive lymphomas as demonstrated by flow cytometry analysis and in situ hybridization. These results highlight the safety of the administration of G^bG^M LV-modified HSC with long-term follow-up after primary and secondary transplants in mice. This data supported the initiation of phase 1/2 first-in-human SCA clinical trial in the United States.