Figure 6.

Persistence of genome edited cells in vivo is greatest in mice transplanted with sequentially dual-edited cells but translocations persist in vivo

(A) HBG-113 total editing within human cells in the infusion product and subsequently the PB of transplanted mice over time (mean ± SD). (B) BCL11A-ee editing within human cells in the infusion product and subsequently the PB of transplanted mice over time (mean ± SD). (C) HBG-113 total editing within human cells in the PB, BM, and spleen at necropsy (mean ± SD). (D) BCL11A-ee editing within human cells in the PB, BM, and spleen at necropsy (mean ± SD). (E) Example of flow cytometric gating strategy for the assessment of HbA and HbF positivity within cells cultured from mouse BM. (F) Assessment of HbF reinduction within BM cells extracted from mice at necropsy and cultured in differentiation media, by flow cytometry and by HPLC (mean ± SD). (G) Gel electrophoretic image of the results of qualitative translocation PCR conducted on mouse PB samples at 6 weeks posttransplant using HBG-113 F and BCL11A-ee R primers. (H) Quantitative assessment of the frequency of translocation events within human cells in the BM of transplanted mice at necropsy (23 weeks posttransplant) by ddPCR (mean). (I) Quantitative assessment of the frequency of translocation events within human cells in the PB and splenic tissue of mice transplanted with dual-edited reactions at necropsy (mean). BCL11A, BCL11A-ee; HBG: HBG-113; Sim, simultaneous; seq, sequential. Only significant differences (p < 0.05) are shown.