Extended Data Fig. 7: PRMT9 inhibition eradicates AML in vivo.
a, Control or inducible Prmt9 KD MA9-lucifase cells were injected into wild-type B6 (1×106 cells per mouse, n = 5/group). After 30 days when leukemia robustly developed, mice were continuously administered with Dox. Kaplan-Meier curves show survival. P value was determined by log-rank (Mantel–Cox) test. b, MA9 AML burden was assessed by bioluminescence imaging over indicated days and the statistics for the quantitative results on day 30 and day 50 from bioluminescence imaging were shown, n = 5 mice/group. Data are presented as mean ± SEM. P values were determined by unpaired two-sided t test. c, MA9 AML cells we implanted into WT mice and evaluated progression: 1) Ctrl (Prmt9 WT), n = 7 mice; 2) Prmt9 KD, n = 7 mice; 3) Prmt9 KD with T cell depletion, n = 5 mice; 4) Prmt9 KD with NK cell depletion, n = 5 mice. Kaplan-Meier curves show survival. P value was determined by log-rank (Mantel–Cox) test. d, following engraftment, 1 day prior to in-vivo DOX administration to KD Prmt9, mice are administered with anti-CD4/CD8 treatment or anti-NK1.1 to deplete T or NK cells. Plot of the depletion in PB was shown. e, Control or Prmt9 KD MA9-lucifase cells were injected into wild-type B6 mice (1×106 cells per mouse, n = 5). Following engraftment, mice were treated with Dox in drinking water for 7 days. The plot shows the frequency of MA9, CD3 + , B220+ and Gr1 + /Mac1+ cells in BM of each mouse at the time of collection analyzed by flow-cytometry. BM and spleen cells from one representative mouse (with the frequency of each subset highlighted in red) in each group were selected and subjected to scRNA-seq analysis. Data are presented as mean ± SEM. P value of each comparison was determined by unpaired two-sided t test. f, identified populations in MA9 BM. g, Frequency of AML cells in BM. h–j, Different populations (h) and markers (i) identified within Ctrl and Prmt9 KD groups merged in spleen cells. identified spleen populations are shown (j). k, T cell frequency in spleen cells. l, Expression of T cell marker genes in spleen. m, Frequency of Cd44+ cells in BM T cells. n, Frequency of Tregs (Foxp3+) in BM CD4 + T cells. o, Plot of leukemic BM in MA9 rechallenge. p, GSEA analysis of BM T cells. Normalized enrichment scores and family-wise error rate P-value was determined by the GSEA permutation method. The Normalized Enrichment Score is calculated by dividing the Enrichment Score from the actual ranking by the means of the random permutations. An enrichment P-value is calculated by comparing the observed frequency of an annotation term with the frequency expected by chance; individual terms beyond cut-off (p-value ≤ 0.05) are deemed enriched. q–u, (q-r) UMAP (q) and histogram (r) showing Isg15 expression in indicated BM subpopulation; (s-t) UMAP and histogram showing Ifit1 expression in indicated BM population; (u) Cxcl10 expression in indicated BM populations. Ctrl: T cells, n = 249 cells; Monocytes/Macrophages, n = 631 cells; DCs, n = 108 cells; Granulocytes, n = 3378 cells; B cells, n = 413 cells. Prmt KD: T cells, n = 231 cells; Monocytes/Macrophages, n = 469 cells; DCs, n = 57 cells; Granulocytes, n = 3906 cells; B cells, n = 134 cells. For (o, q, r), Data were mean ± SEM. P values were determined by unpaired two-sided t-tests.
