Fig. 4: NK cells from HDs efficiently target CCUS mutant cells.

A Frequencies of MDS-L cells in the BM (left) and spleen (right) of engrafted NSGS mice 10 days after the injection of NK cells transfected with Cas9 in the absence (Cas9; n = 8) or presence of sgRNAs against TET2 (TET2 KO; n = 6) or DNMT3A (DNMT3A KO; n = 6). Statistical significance was calculated using one-way ANOVA and Tukey’s multiple comparisons test. **P < 0.01, ****P < 0.0001. BM, TET2 KO vs DNMT3A KO: P = 0.0035; spleen, TET2 KO vs DNMT3A KO: P = 0.0084. hCD45, human CD45. Error bars represent the mean ± standard deviation. B PB blood counts in engrafted NSGS mice 10 days after the injection of NK cells transfected with Cas9 (n = 8) in the absence or presence of sgRNAs against TET2 (n = 6) or DNMT3A (n = 6). Dotted lines represent the mean of hemoglobin, white blood cell counts, and platelets for 8 week-old NSGS mice based on the Mouse Phenome Database (Jackson Laboratories). Statistical significance was calculated using one-way ANOVA and Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ****P < 0.0001. Hemoglobin, Cas9 vs DNMT3A KO: P = 0.004; TET2 KO vs DNMT3A KO: P = 0.0651. White blood cells (WBC), Cas9 vs TET2 KO: P = 0.005; Cas9 vs DNMT3A KO: P = 0.0103; TET2 KO vs DNMT3A KO: P = 0.9445. Platelets, TET2 KO vs DNMT3A KO: P = 0.1041. Error bars represent the mean ± standard deviation. C Kaplan–Meier survival analysis of NSGS mice transplanted with MDS-L cells injected with vehicle (n = 5) or NK cells transfected with Cas9 in the absence (n = 8) or presence of sgRNAs targeting TET2 (n = 8) or DNMT3A (n = 8). Statistical significance was calculated using a log-rank (Mantel-Cox) analysis adjusted by Holm’s multiple comparisons test. ***P < 0.001. Vehicle vs Cas9: P = 0.0002; Vehicle vs TET2 KO: P = 0.0002; Vehicle vs DNMT3A KO: P = 0.0002; TET2 KO vs DNMT3A KO: P = 0.2036. D Frequencies of human CD45⁺CD56^- cells in the BM of MISTRG mice transplanted with cells from 4 patients with CCUS. Samples were analyzed 1 week after the injection of vehicle (n = 3/each) or HD NK cells (n = 3/each). Dotted lines are color-coded and represent the mean of each group. Statistical significance between treatment groups was calculated using a two-tailed nested t-test. *P < 0.05. Vehicle vs NK cells: P = 0.0486. Error bars represent the mean ± standard deviation. E VAF in mouse CD45-depleted and human CD45-enriched BM cells isolated from MISTRG donor mice transplanted with cells from 2 CCUS patients 1 week after the injection of vehicle (n = 3/each) or healthy NK cells (n = 3/each). Dotted lines are color-coded and represent the mean of each group. Statistical significance between treatment groups was calculated using a two-tailed nested t-test. *P < 0.05. Vehicle vs NK cells: P = 0.0279. Error bars represent the mean ± standard deviation. Source data are provided as a Source Data file.