Extended Data Fig. 6: Efficient HSPCs editing by LNP-028-ABE8e-HBG in immunodeficient mice.
a-c. BM was collected two weeks after the second-dose and fifth-dose injections of LNP-028-ABE8e-HBG, and then analyzed by flow cytometry for human cell chimerism (a), multilineage reconstitution (b), or human erythroid cells (c) in the BM. d. Base editing efficiency was assessed in vitro using differentiated erythroid cells derived from the BM cells of engrafted mice after injection of LNP-028-ABE8e-HBG. e. The γ-globin expression was analyzed by RT-qPCR in erythroid cells that were differentiated in vitro from BM cells edited by LNP-028-ABE8e-HBG. f. Enucleation of in vitro differentiated erythroid cells from LNP-028-ABE8e-HBG edited BM cells. For figs. a-f, each symbol represents a mouse, and there were a total of n = 3 mice. g. After LNP-028-ABE8e-HBG edited BM cells were transplanted into NCG-X mice, the chimerism level of human cells among the transplanted BM cells was analyzed by flow cytometry 16 weeks post-transplantation. h. The percentage of engrafted human B cells, myeloid cells, and CD19-CD33- cells among the transplanted BM cells was assessed after 16 weeks of transplantation using flow cytometry (n = 5 mice). i. After 16 weeks of transplantation, deep sequencing analysis was conducted to evaluate the base editing efficiency at positions A5 and A6 within different hematopoietic cell populations derived from the engrafted BM cells (n = 4 mice). All statistical significances in the figures were analyzed using one-way ANOVA with Dunnett’s multiple comparisons test, and data represent mean ± SD. Statistical analysis showed NS for second- and fifth-dose groups compared with the un-edited control in panels a and c. Panel d also showed NS between non-mobilized and mobilized groups, but p < 0.0001 was observed for both doses. Panel e confirmed p < 0.0001 for second- and fifth-dose, while panel g showed NS for the same comparison.
