Fig. 7: ABT-263 targets a senescent macrophage signature in MASLD.
a, Model illustration of CETP-APOE* Leiden transgenic mice maintained on C57BL/6J then crossed to three additional genetic backgrounds susceptible (BXD19/TyJ and C57BL/6J) and resistant (129/SvJ) to developing to MASLD. b, Representative picrosirius red stain for liver fibrosis after 16 weeks on HFHCD. Scale bars represent a 20 μm distance. c, Pathology grades of fibrosis of n = 4–6 male liver sections stained with picrosirius red stain. Data represent median ± s.e.m. d, Bulk RNA-seq of transcript levels for MSen gene signature across weeks 0–16 on HFHCD. e, Mean ± s.e.m. MSen scores across weeks 0–16 on HFHCD for each genetic background. Best-fit line and P value derived from a simple linear regression model of n = 3–6 male mice per time point. f, Mean ± s.e.m. mRNA transcript levels (RT–qPCR) relative to control of BMDMs from CETP-APOE*Leiden mice crossed on different three backgrounds. P value from a two-way ANOVA. n = 3 biological replicates per strain and condition were used. g, Model illustration of CETP-APOE*Leiden transgenic mice crossed on the C57BL/6J genetic background. Mice were treated with ABT-263 via oral gavage every day for 7 days on weeks 12 and 15. h, Mean ± s.e.m. weight of n = 9 placebo mice and n = 10 ABT-263-treated mice at 17 weeks. P value derived from a two-sided t-test. i, Mean ± s.e. change in food intake for n = 6 placebo cages and n = 5 ABT-263-treated cages at 17 weeks. j, Mean ± s.e. spleen weight (in grams) relative to total weight of n = 9 placebo mice and n = 10 ABT-263-treated mice at 17 weeks. P value derived from a two-sided t-test. k, Enzyme-linked immunosorbent assay quantification for picograms of tumor necrosis factor found in every microliter of serum collected. P value derived from a one-sided t-test. Mean ± s.e.m. of n = 9 placebo mice and n = 10 ABT-263-treated mice at 17 weeks. l, Mean liver weight (in grams) relative to total weight ± s.e. of n = 9 placebo mice and n = 10 ABT-263-treated mice at 17 weeks. P value derived from a two-sided t-test. m, Representative liver photographs at 17 weeks immediately after tissue harvest. Scale rule: inches. n, Mean mRNA transcript levels (RT–qPCR) for SASP, senescent and macrophage genes in bulk liver samples relative to placebo control. P value derived from a two-sided t-test. Bar plots: average ± s.e.m. of n = 9 placebo mice and n = 10 ABT-263-treated mice at 17 weeks. o, Bulk RNA-seq of the MSen transcriptomic signature (genes derived from aged macrophages in vivo and senescent macrophages in vitro) in response to the senolytic drug, ABT-263. Scale bar: log2[TPM + 1] expression on a z-score axis. n = 6 vehicle and n = 5 ABT-263-treated mice used in this experiment. p, GSVA scoring of MSen genes in the livers of vehicle- or drug-treated mice. Box-and-whisker plots display the median (center line), interquartile range (25th to 75th percentiles; box), and minimum and maximum values (whiskers). P values were calculated using a nonparametric Wilcoxon test. n = 6 vehicle-treated and n = 5 ABT-263-treated mice were analyzed. q, Total NAD (pmol per mg of liver analyzed) quantified by LC–MS/MS. Mean ± s.e.m. of NAD distribution in response to ABT-263. n = 9 placebo mice and n = 10 ABT-263-treated mice at 17 weeks. P value derived from a two-sided t-test. r, Nonalcoholic fatty liver disease (NAFLD) activity score based on pathology grades for each condition. Bar: mean ± s.e. P value derived from Student’s t-test. s, Oil Red O stains are shown for three representative mice treated with vehicle or ABT-263. Scale bars represent a 20 μm distance. Mean ± s.e.m. lipid droplet size and area analyzed in eight images per liver section, and the average size (μm2) from eight slides for each mouse was used for statistical analysis. n = 6 vehicle and n = 5 ABT-263-treated mice used in this analysis. P value derived from Student’s t-test.
