Fig. 2: Succinate promotes acute myeloid leukemia progression in mouse models with low SUCNR1 expression.
From: Succinate receptor 1 restricts hematopoiesis and prevents acute myeloid leukemia progression
A Droplet digital (dd)PCR expression of Sucnr1 relative to Gapdh in bone marrow nucleated cells (BMNC) from control (Mx1-Cre− NRASG12D, n = 3) and NRAS-G12D+ (Mx1-Cre+ NRASG12D, n = 4) mice, 11 weeks after poly-inosine:poly-cytosine (pIpC). B Intracellular succinate levels in total BM (TBM) Lin−Sca-1+c-Kit+ (LSK) cells and CD11b+Gr-1+F4/80+ monocytes obtained by fluorescence-activated cell sorting (FACS)-sorting from control (n = 3 Mono, n = 5 LSK) and NRAS-G12D+ (n = 3 Mono, n = 6 LSK) mice, 5 weeks after pIpC. C Succinate levels in BM extracellular fluid (BMEF) from control and NRAS-G12D+ mice measured by colorimetric assay 5 weeks after pIpC induction (n = 3 per group, left) and by nuclear magnetic resonance 6 weeks after pIpC induction (n = 4 per group, right). D–F Treatment with succinic acid (Succ. ac.) or vehicle (Veh) of NRAS-G12D+ mice for 27 weeks, starting 7 weeks after pIpC induction. TBM fractions analyzed by FACS of D CD11b+ myeloid cells and B220+ lymphocytes (n = 7 per group), (E) LSK cells (n = 5 Veh, n = 7 Succ. ac.), F LSK cell subsets: LSK CD34−Flt3−CD150+CD48−, hematopoietic stem cells (HSC); LSK CD34+Flt3−CD150+CD48−, multipotent progenitors 1 (MPP1); LSK CD34+Flt3−CD150+CD48+ (MPP2); LSK CD34+Flt3−CD150−CD48+ (MPP3); LSK CD34+Flt3+CD150−CD48+ (MPP4); LSK CD34+Flt3−CD150−CD48− (MPP5); LSK CD34−Flt3−CD150−CD48− (MPP6) (n = 5 Veh, n = 7 Succ. ac.). G ddPCR expression of Sucnr1 relative to Gapdh in BMNC from wild-type (WT) and Mll-AF9 mice (n = 3 per group). H–J Treatment with succinic acid or vehicle of C57BL6/J mice transplanted with BMNC from leukemic Mll-AF9 mice for a maximum of 42 weeks, starting 4 weeks after transplant. H Probability of survival of recipient mice treated with succinic acid (n = 5) or vehicle (n = 8) with leukemia-confirmed deaths, calculated by Kaplan–Meier method. I TBM fractions of Lin− cells analyzed by FACS at endpoint (n = 7 Veh, n = 5 Succ. ac.). J TBM and total spleen (TSP) cell fractions of monocytes analyzed by FACS (n = 8 Veh, n = 5 Succ. ac.). Statistical analyses were performed with two-tailed Student’s t-test (A–G, I, J) or log-rank Mantel-Cox test (H). Significant p-values are reported. Data are biologically independent animals, and means ± SEM for bar plots. Source data and non-significant p-values are provided as a Source Data file.
