Fig. 5: Genetic and pharmacological inhibition of IL1R1 abrogates aberrant myeloid expansion in Tet2-KO mice.

a–d WT, Vav-cre Tet2^fl/fl (Tet2-KO), Il1r1-/- (Il1r1-KO), and Vav-cre Tet2^fl/flIl1r1^-/- (double knockout; DKO) mice were bled twice and 8 mice were sacrificed at greater than 60 weeks of age, the remaining cohort was followed to observe survival outcomes. a Experimental design. b Spleen weight in grams. c Survival of WT (n = 8), Tet2-KO (n = 16), Il1r1-KO (n = 15), and DKO (n = 18) mice. d The frequency of MHCII⁺ macrophages (CD45⁺CD11b⁺F4/80⁺MHCII⁺), F4/80^- Ly6c^hi monocytes (CD45⁺CD11b⁺F4/80^-Ly6c⁺Ly6g^-) and F4/80^- Ly6c^lo monocytes (CD45⁺CD11b⁺F4/80^-Ly6c^-Ly6g^-) in BM and spleen. e–g Lineage-depleted BM cells derived from WT CD45.1 and Tet2-KO CD45.2 mice were transplanted into lethally irradiated WT heterozygous CD45.1/2 mice. Three weeks after the transplantation mice were treated with IL1β (500 ng/mouse/day) or vehicle as well as with or without an IL1R1 antagonist (anakinra, 100 mg/kg) (n = 3 mice/group for Veh, Ana, and IL1β, and 4 for IL1β Ana). e Experimental design, f the percentage of Tet2-KO CD45.2 donor cells out of all CD45⁺ cells (chimerism) in PB, and g the frequency of LK and LSK cells in the spleen. h Human BM CD34⁺ progenitors were CRISPR-Cas9 edited for TET2 (TET2-sgRNA) alongside non-targeting control (NT-sgRNA), then 600 cells per well were plated with or without IL1β (25 ng/mL) and/or IL1RA (50 ng/mL). Colonies were counted after 2 weeks and individual colonies were picked for sanger sequencing to determine indel frequency within TET2. Total colony numbers at 2 weeks and CFU-M (monocyte) colony numbers, with the number of colonies identified with detectable indels shown. Error bars represent mean ± SEM. For panels a, d, g, f, and h two-factor ANOVA was used to determine the FWER-adjusted p values. For panel f, a student’s two-tailed t-Test was used to determine significance. For FWER adjusted and regular p values: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.