Figure 4: Correction of anaemia in thalassemic mice by NPs containing γtcPNA and donor DNA. | Nature Communications

Figure 4: Correction of anaemia in thalassemic mice by NPs containing γtcPNA and donor DNA.

From: In vivo correction of anaemia in β-thalassemic mice by γPNA-mediated gene editing with nanoparticle delivery

Figure 4

(a) Blood smears from wild-type and thalassemic mice obtained pre-treatment or 36 days after in vivo treatment with blank NPs, SCF alone, SCF plus scrambled γtcPNA4-Scr/donor DNA NPs or SCF plus γtcPNA4/donor DNA NPs. NPs were given i.v.; SCF i.p. The untreated group (and control animals) exhibit extreme poikilocytosis as well as numerous target cells, cabot rings, anisochromasia and ovalocytosis, all changes characteristic of β-thalassemia. Treatment with γtcPNA4/donor DNA and SCF ameliorates the poikilocytosis and yields a reduction in anisocytosis, ovalocytosis and target cells suggestive of reduced alpha-globin precipitation in the RBCs. Scale bar, 1.0 μm. (b) Blood haemoglobin levels of thalassemic mice treated with blank NPs, SCF plus scrambled γtcPNA4-Scr/donor DNA NPs, or with SCF plus γtcPNA4/donor DNA NPs performed at the indicated number of days after treatment, up to 140 days. Data are presented as box and whisker plots showing the median and quartile range within each group over time (n=6 per group). Only the SCF plus γtcPNA4/donor DNA-treated mice achieved and maintained haemoglobin levels within the normal range during the duration of the experiment, reflecting the increased haemoglobin stability conferred by the gene editing. Horizontal bars within the boxes indicate mean; statistical analysis by Student’s unpaired t-test, *P<0.05. (c) Reticulocyte counts (% of total RBCs) calculated in blood smears from thalassemic mice treated with either blank NPs or with NPs containing and γtcPNA4/donor DNA plus SCF on days 0 and 36 post treatment. Data are mean±s.e.m., n=3; statistical analysis by Student’s t-test, *P<0.05. (d) Images of spleens from wild-type mice or thalassemic mice treated with blank NPs, SCF alone, SCF plus scrambled γtcPNA4-Scr/donor DNA NPs or SCF plus γtcPNA4/donor DNA NPs at 36 days after the last treatment. Average spleen weights (with standard errors, n=3; except for wild-type, where n=1) are listed below the images. Splenomegaly is corrected only by SCF plus γtcPNA4/donor DNA treatment. Scale bar, 1.0 cm. (e) Histopathologic analysis of spleen sections from wild-type mice and from thalassemic mice obtained 36 days after treatment with blank NPs or SCF plus γtcPNA4/donor DNA NPs. Haematoxylin and eosin stain (H&E), × 40 magnification. Scale bar, 10.0 μm. Additional images of spleens from other treatment groups and with additional stains are presented in Supplementary Figs 17 and 18.

Back to article page