Mutation-dependent responses to sleep and exercise in clonal haematopoiesis

Abstract Clonal haematopoiesis (CH) activates inflammation and increases the risk of atherosclerosis1,2. Whether lifestyle alters CH clone expansion or the phenotypic programming of CH mutant cells, thereby affecting atherosclerosis, is unknown. Here, in humans and mice and across mutations in Jak2, Tet2, Trp53 and Dnmt3a, we demonstrate mutation-dependent responses to sleep and exercise in CH and show that mutant cells are uniquely sensitive to lifestyle. In two human datasets, moderate-to-vigorous physical activity was associated with lower prevalence of non-DNMT3A-driven CH. In atherogenic mice with Jak2V617F or Tet2 loss of function (LOF), but not Trp53 LOF or Dnmt3aR878H CH, uninterrupted sleep or exercise curtails clone expansion. In CH with the Jak2V617F mutation, sleep and exercise reduces clone expansion by selectively reprogramming mutant, but not cohabitant wild type, haematopoietic progenitor cells towards antiproliferative and metabolically healthy phenotypes by tempering bone marrow macrophage–haematopoietic progenitor cell IL-1β signalling. Sleep or exercise also lessens Jak2V617F-driven, Tet2 LOF-driven and Trp53 LOF-driven, but not Dnmt3aR878H-driven, atherosclerosis by locally reprogramming mutant vascular macrophages, independent of peripheral clone dynamics. In Jak2V617F, but not adjacent wild type, aortic macrophages, uninterrupted sleep blunts CLEC4E-dependent inflammasome activation, consequently diminishing lesions. Exercise, meanwhile, activates PAC1+ neurons in the locus coeruleus, raising the levels of peripheral noradrenaline, which signals through adrenergic receptor β2 (ADRβ2) whose expression is preserved by exercise in Jak2V617F, but not cohabitant wild type, aortic macrophages, selectively repressing their inflammatory programming and atherosclerosis. Our findings establish that healthy lifestyles gene-specifically diminish CH and selectively reprogram mutant haematopoietic progenitor cells and macrophages to maintain cardiovascular health. 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Funding This work was supported in part through the Minerva computational and data resources and staff expertise provided by Scientific Computing and Data at the Icahn School of Medicine at Mount Sinai, and supported by the Clinical and Translational Science Awards (CTSA) grant UL1TR004419 from the National Center for Advancing Translational Sciences. This work was funded by a Walter Benjamin fellowship by the German Research Foundation DFG GE 3588/1-1 and by the Work–Family Balance Funding Program of the German Society of Internal Medicine (DGIM; both to T.G.); an MD fellowship from the Boehringer Ingelheim Fonds (to M.H.); an American Heart Association postdoctoral fellowship 24POST1196847 (to P.H.); NIH T32MH087004 (to P.T.); a research fellowship from the Belgian American Educational Foundation (to D.E.); NIH K23HL169839 and AHA 23CDA1050571 (to S.K.); NIH K08HL166687, R01HL173028 and the American Heart Association 24RGRSG1275749, 25SFRNPCKMS1463898 and 25SFRNCCKMS1443062 (to M.C.H.); NIH R01HL158534, R01AG082185, R01HL178835 and R00HL151750, and the American Heart Association 25TPA1477871, the Cure Alzheimer’s Fund and the Alzheimer’s Association (to C.S.M.). Author information Authors and Affiliations Contributions T.G., W.J., L. Gaebel and M.H. conceived the project, designed and performed experiments, analysed and interpreted the data, offered intellectual input and edited the manuscript. C.W., P.H., B.G.D.S., P.T., N.F.B., A.G.Y., A.K., A.D., M.G., M.G.K., N.Y. and S.G. conducted and aided the experiments and data acquisition. T.F. and O.C. interpreted the data and offered intellectual input. D.N., R.C., D.D.’S., Z.C., E.R. and S.K.-S. conducted the sequencing experiments and aided in analysis. F.K.S., A.K.R., M.M. and L. Goedeke aided in supervision and offered intellectual input. T.N., M.M.U., S.K., A.G.B., P.N., P.T.E., D.E. and M.C.H. performed, aided and supervised the human data analysis. C.S.M. conceived the project, supervised, directed and managed the study, interpreted data, designed the figures and wrote the manuscript. Corresponding author Ethics declarations Competing interests C.S.M. is a consultant for Granite Bio, unrelated to the present work. M.C.H. reports consulting fees from Comanche Biopharma; site principal investigator work and advisory board service for Novartis; and research support from Genentech, unrelated to the present work. S.K. has received sponsored research support from Bayer AG, unrelated to the present work. P.N. reports research grants from Allelica, Amgen, Apple, Boston Scientific, Cleerly, Genentech/Roche, Ionis, Novartis and Silence Therapeutics; personal fees from Allelica, Apple, AstraZeneca, Bain Capital, Blackstone Life Sciences, Bristol Myers Squibb, Creative Education Concepts, CRISPR Therapeutics, Eli Lilly & Co, Esperion Therapeutics, Foresite Capital, Foresite Labs, Genentech/Roche, GV, HeartFlow, Magnet Biomedicine, Merck, Novartis, Novo Nordisk, TenSixteen Bio and Tourmaline Bio; equity in Bolt, Candela, Mercury, MyOme, Parameter Health, Preciseli and TenSixteen Bio; and spousal employment at Vertex Pharmaceuticals, all unrelated to the present work. The other authors declare no competing interests. Peer review Peer review information Nature thanks Hafid Ait-Oufella, Rachel Rowe and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available. Additional information Extended data figures and tables Extended Data Fig. 1 Supplemental analysis of UK Biobank and All of Us datasets, exercise and sleep phenotypes in CH mice. a. Flow chart for UK Biobank and All of Us datasets. b. Meta-analysis of the association of moderate-to-vigorous physical activity with the prevalence of DNMT3A CH and non-DNMT3A CH, split by sex. Schematic created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). c. Quantification of distance run per hour over 24 h by Ldlr−/−WTBM, Ldlr−/−Jak2V617FBM, Ldlr−/−Tet2−/−BM, Ldlr−/−p53−/−BM, and Ldlr−/−Dnmt3aR878HBM mice undergoing exercise at baseline, 4, 8, and 12 weeks (n = 4, 4, 4, 4; 3, 4, 4, 4; 5, 5, 5, 5; 5, 5, 4, 5 and 2, 5, 7, 4 at baseline, 4, 8 and 12 weeks, respectively). d.e. EEG/EMG quantification of slow-wave sleep and wake bout frequency 10 weeks after BM transplant inLdlr−/−WTBM, Ldlr−/−Jak2V617FBM, Ldlr−/−Tet2−/−BM, Ldlr−/−p53−/−BM, and Ldlr−/−Dnmt3aR878HBM mice in (d) a sedentary state and (e) during sleep fragmentation (n = 5 Ldlr−/−WTBM, n = 5, 5; 4, 4; 4, 3 and 3, 3 for Sedentary vs SF. P values are derived from independent Welch’s T Tests. Two-sided statistics. Data are mean ± s.e.m; * p < 0.05. Extended Data Fig. 2 Extended immunophenotyping of CH mice exposed to SF and exercise. a. Flow cytometry gating strategy to identify CD45.2 mutant or control and CD45.1 wild-type Ly6Chi monocytes and neutrophils in blood. b. Engraftment frequencies of CD45.2 and CD45.1 Ly6Chi monocytes and neutrophils in the blood of Ldlr−/−WTBM and Ldlr−/−Jak2V617FBM mice at baseline (n = 10 wt, 18 sed, 16 SF, 16 exercise) and total cell counts at the time of euthanasia (n = 8-10 wt, 13-14 sed, 12-13 SF, 6-8 exercise), with representative flow cytometry plots. c. Plasma IL-6 levels at time of euthanasia in Ldlr−/−WTBM and Ldlr−/−Jak2V617FBM mice (n = 5 wt, 5 sed, 6 SF, 6 exercise). d–f. Body weight (d), plasma lipids (e), and blood parameters (f) in Ldlr−/−WTBM and Ldlr−/−Jak2V617FBM mice exposed to lifestyle at the time of euthanasia (5, 4, 5-6 wt; 6, 5-6, 8-9 sed; 10, 10, 5 SF and 6, 6, 6 exercise for body weight, plasma lipids and blood parameters, respectively). g. Analysis of CD45.2 Ly6Chi monocyte and CD45.2 neutrophil fraction growth in Ldlr+/+ mice with WTBM or Jak2V617F CH exposed to SF or exercise (n = 4, 4 wt; 4, 4 sed; 4, 5 SF and 2, 2 exercise). h. Engraftment frequencies of CD45.2 and CD45.1 Ly6Chi monocytes and neutrophils in the blood of Ldlr−/−WTBM and Ldlr−/−Tet2−/−BM mice at baseline (n = 7 wt, 7 sed, 6 SF, 6 exercise) and total cell counts at the time of euthanasia (n = 5-6 wt, 7 sed, 7 SF, 5-6 exercise), with representative flow cytometry plots. i. Plasma IL-6 levels at time of euthanasia in Ldlr−/−WTBM and Ldlr−/−Tet2−/−BM mice (n = 6 wt, 7 sed, 8 SF, 5 exercise). j–l. Body weight (j), plasma lipids (k), and blood parameters (l) in Ldlr−/−WTBM and Ldlr−/−Tet2−/−BM mice exposed to lifestyle at the time of euthanasia (6, 4-6, 2 wt; 7, 7, 2 sed; 7, 7, 2 SF and 6, 6, 3 exercise for body weight, plasma lipids and blood parameters, respectively). m. Engraftment frequencies of CD45.2 and CD45.1 Ly6Chi monocytes and neutrophils in the blood of Ldlr−/−WTBM and Ldlr−/−p53−/−BM mice at baseline (n = 5 wt, 9 sed, 10 SF, 12 exercise) and total cell counts at the time of euthanasia (n = 4 wt, 9 sed, 9-10 SF, 7-8 exercise) with representative flow cytometry plots. n. Plasma IL-6 levels at time of euthanasia in Ldlr−/−WTBM and Ldlr−/−p53−/−BM mice (n = 6 wt, 4 sed, 9 SF, 3 exercise). o.p. Body weight (o) and plasma lipids (p) in Ldlr−/−WTBM and Ldlr−/−p53−/−BM mice exposed to lifestyle at the time of euthanasia (3, 4 wt; 9, 9-10 sed; 10, 9 SF and 9, 8 exercise for body weight and plasma lipids, respectively). q. Engraftment frequencies of CD45.2 and CD45.1 Ly6Chi monocytes and neutrophils in the blood of Ldlr−/−WTBM and Ldlr−/− Dnmt3aR878HBM mice at baseline (n = 6 wt, 7-8 sed, 8 SF, 8 exercise). Analysis of CD45.2 Ly6Chi monocyte and CD45.2 neutrophil fraction growth in Ldlr−/−WTBM or Ldlr−/− Dnmt3aR878HBM mice undergoing lifestyle intervention (n = 6 wt, 10 sed, 7 SF, 8 exercise) with representative flow cytometry plots. Time course of plasma IL-1β levels (n = 6 wt, 11 sed, 8 SF, 8 exercise). r. Body weight and plasma lipid levels in Ldlr−/−WTBM and Ldlr−/− Dnmt3aR878HBM mice exposed to lifestyle at the time of euthanasia (4, 4 wt; 6, 6 sed; 7, 7, SF and 5, 5 exercise). Two-sided statistics. Data are mean ± s.e.m; * p < 0.05, ** p < 0.01. P-values were obtained from independent, two-tailed, Welch’s t-tests (for parametric data) or Mann–Whitney U tests (for non-parametric data), 2-way ANOVA + Tukey’s multiple comparisons test (g). Mixed-effects analyses were performed to obtain a group effect statistic for blood expansion curves in (q). Schematics in panels a–c,e,f,g,i,k,l–n,p–r created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). Extended Data Fig. 3 Analysis of SF and exercise in Ldlr−/− mice without CH. a. Experimental setup. b. Plasma levels of IL-1β and IL-6 at the time of euthanasia (n = 5, 4 sed; 6, 3 SF; 6, 5 exercise mice). c. Blood counts of Ly6Chi monocytes and neutrophils at the time of euthanasia (all n = 6 mice). Schematics in panels a–c created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). d. Atherosclerotic lesion area and volume as determined by Oil-red-O positive area of sequential aortic root sections (all n = 6). One representative image is shown per group. e. Proportion of necrotic core as measured by Masson’s Trichrome Staining (n = 5 sed, 6 SF, 6 exercise mice). One representative image is shown per group. f. Immunofluorescence staining and quantification of atherosclerotic plaques according to lifestyle after 16 weeks of high cholesterol diet. Sections were stained for IL-1β, AIM2, and NLRP3, with Mac-2 as a counterstain to identify macrophages (all n = 6 mice). Representative images are shown for each condition. g. Proportion of lesion macrophages (right) that express Clec4e as determined by immunofluorescence (all n = 6 mice). All scale bars are 400 µm. Two-sided statistics. Data are mean ± s.e.m; ns = not significant, * p < 0.05, ** p < 0.01,**** p < 0.0001. d: p-values were derived from a 2-way ANOVA, followed by a Tukey’s multiple comparison test; all other p-values were obtained from independent, two-tailed, Welch’s t-tests (for parametric data) or Mann–Whitney U tests (for non-parametric data). Extended Data Fig. 4 Bone marrow progenitor analysis in CH mice exposed to SF or exercise. a. Flow cytometry gating strategy to identify CD45.2 mutant or control and CD45.1 wild-type progenitor populations in bone marrow. b–e. Cell counts of CD45.2 mutant or CD45.1 wild-type LSKs, CMPs, GMPs, and MDPs in Ldlr−/−Jak2V617FBM (b), Ldlr−/−Tet2−/−BM (c), Ldlr−/−p53−/−BM (d), and Ldlr−/−Dnmt3AR878HBM (e) mice. Proliferation of cell populations, quantified by the proportion of ki67-positive cells using flow cytometry. n = 3-5 wt, 6-7 sed, 8-10 SF, 5-6 exercise. (b), 5-6 wt, 6-7 sed, 6-7 SF, 6 exercise (c), 3-4 wt, 8-9 sed, 9-10 SF, 7-8 exercise (d), and 5-7 sed, 5-7 SF, 4-5 exercise (e). BM levels of IL-6 at time of euthanasia (n = 4, 4, 4 wt; 5, 4, 7 sed; 5, 4, 9 SF and 6, 4, 6 exercise for Ldlr−/−Jak2V617FBM, Ldlr−/−Tet2−/−BM and Ldlr−/−p53−/−BM mice, respectively). Two-sided statistics. Data are mean ± s.e.m; ns = not significant, * p < 0.05, ** p < 0.01. P-values were obtained from independent, two-tailed, Welch’s t-tests (for parametric data) or Mann–Whitney U tests (for non-parametric data). Schematics in panels a–e created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). Extended Data Fig. 5 Extended mechanistic analysis linking sleep and exercise to BM clonal dynamics. a. t-SNE visualization of bone marrow scRNAseq after exclusion of minor clusters <100 cells; n = 5 pooled mice per group. b. Proliferation (Ccnd2, Ccnd3, Pim1, Npm1, Myh9, Dnmt1; left) and metabolism (Atp5k, mt-Nd3, Atp5md, Ndufb1-ps, Ndufa3, Ass1, Mospd3; right) feature scores in wild type CD45.1 and CD45.2 myeloid progenitors; n = 5 pooled mice per group. c. Heat map of select proliferation-associated gene expression in Jak2V617F myeloid progenitor cells from sedentary and exercised mice. d. Experimental setup and mitochondrial respiration marked by OCR in BMDMs isolated from Ldlr−/−Jak2V617FBM mice and sorted into mutant CD45.2 and non-mutant CD45.1 groups; n = 8 and 6 technical replicates for the sedentary CD45.1 and CD45.2 groups; n = 12 and 8 technical replicates for the CD45.1 and CD45.2 exercise groups. e. UMAP from bone marrow single-cell RNA sequencing data integrating myeloid CD45.2 progenitors from all four conditions, overlaid with a composite gene score for Il1r1 and Il1rap. f. Representative flow cytometry plots showing pro-IL-1β expression in various cell types from the BM of Ldlr−/− Jak2V617FBM mice. g. Experimental setup, and representative flow cytometry plots showing ki67 expression in bone marrow CMPs in WTJak2V617FBM mice injected with saline or IL-1β, as well as quantification of %ki67 positivity in CMPs. n = 3 for saline groups, n = 5 for IL-1β groups. h. Bodyweight, plasma cholesterol, and plasma triglyceride levels in Ldlr−/−Jak2V617FBM mice with and without SF, with IgG or anti-IL-1β at the time of euthanasia. n = 10, 10, 9 no SF + IgG; 7, 6, 6 no SF + anti-IL-1β; 13, 13, 13 SF + IgG and 8, 7, 7 SF + anti-IL-1β for bodyweight, cholesterol and triglycerides, respectively. i. Flow cytometry plots for ki67 in CMPs from Ldlr−/−Jak2V617FBM mice with and without SF, with IgG or anti-IL-1β. Two-sided statistics. Data are mean ± s.e.m; ns= not significant, *p < 0.05, **p < 0.01,****p < 0.0001. P-values were derived from ordinary one-way ANOVA tests. Schematics in panels d–i created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). Extended Data Fig. 6 Analysis of aortic macrophage proliferation and recruitment, and atherosclerosis in Ldlr−/−Dnmt3aR878HBM mice exposed to SF or exercise. a. Counts for all macrophages and CD45.1 WT macrophages in the aorta of Ldlr−/−Jak2V617FBM and Ldlr−/−WTBM mice measured by flow cytometry at the time of euthanasia (n = 5, 5 wt; 6, 6 sed; 10, 11 SF and 6, 6 exercise). b. CD45.2+ aortic macrophage proliferation measured by flow cytometry in Ldlr−/−Jak2V617FBM and Ldlr−/−WTBM control mice at the time of euthanasia (n = 5 wt, 7 sed, 11 SF, 6 exercise). c. Representative flow cytometry plots for bead uptake into aortic macrophages in Ldlr−/−Jak2V617FBM and Ldlr−/−WTBM control mice with quantification of percent of bead-positive macrophages across all lifestyle groups (n = 5, 6 wt; 5, 5 sed; 6, 6 SF and 7, 7 exercise for CD45.1 and CD45.2, respectively). d. Correlation between the fractions of CD45.2 monocytes in the blood and CD45.2 macrophages in the aorta of Ldlr−/−Jak2V617FBM and Ldlr−/−WTBM control mice at the time of euthanasia (n = 41, 41 for blood and aorta, respectively). e. Counts for all macrophages and CD45.1 WT macrophages in the aorta of Ldlr−/−Tet2−/−BM and Ldlr−/−WTBM mice measured by flow cytometry at the time of euthanasia (n = 6, 6 wt; 7, 7 sed; 6, 7 SF and 6, 6 exercise). f. CD45.2+ aortic macrophage proliferation measured by flow cytometry in Ldlr−/−Tet2−/−BM and Ldlr−/−WTBM control mice at the time of euthanasia (n = 6 wt, 7 sed, 7 SF and 6 exercise). g. Quantification of bead uptake into aortic macrophages in Ldlr−/−Tet2−/−BM mice across all lifestyle groups (4, 4 sed; 5, 5 SF and 4, 4 exercise). h. Correlation between the fractions of CD45.2 monocytes in the blood and CD45.2 macrophages in the aorta of Ldlr−/−Tet2−/−BM and Ldlr−/−WTBM control mice at the time of euthanasia (n = 26, 26 for blood and aorta, respectively). i. Counts for all macrophages and CD45.1 WT macrophages in the aorta of Ldlr−/−p53−/−BM and Ldlr−/−WTBM mice measured by flow cytometry at the time of euthanasia (n = 4, 4 wt; 8, 8 sed; 9, 8 SF and 8, 8 exercise). j. Quantification of bead uptake into aortic macrophages in Ldlr−/− p53−/−BM mice across all lifestyle groups (4, 4 sed; 5, 5 SF and 4, 4 exercise for CD45.1 and CD45.2, respectively). k. Correlation between the fractions of CD45.2 monocytes in the blood and CD45.2 macrophages in the aorta of Ldlr−/−p53−/−BM and Ldlr−/−WTBM control mice at the time of euthanasia (n = 29, 29 for blood and aorta, respectively). l. Lesion size, volume and necrotic core area (n = 5 wt, 6 sed, 6 SF, 7 exercise) with representative images of aortic roots stained with Masson’s Trichrome stain, quantification of total and CD45.2 aortic macrophage number, and CD45.2 macrophage proliferation in Ldlr−/−Dnmt3aR878HBM and Ldlr−/−WTBM control mice at the time of euthanasia (n = 4 wt, 7 sed, 7 SF, 4 exercise); scale bar=200 µm. Two-sided statistics. Data are mean ± s.e.m; *p < 0.05, **p < 0.01. P-values were obtained from independent, two-tailed, Welch’s t-tests (for parametric data) or Mann–Whitney U tests (for non-parametric data). Schematics in panels a,c,e,i–l created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). Extended Data Fig. 7 Extended scRNAseq analysis of aortic leukocytes. a. Monocyte/macrophage signature genes across all aortic cell types. b. Top 30 cluster-defining genes for CD45.2 macrophage clusters 1-4. c. UMAP of Il1b expression across all aortic leukocyte populations with Log2 expression of Il1b in CD45.1 and CD45.2 macrophages and neutrophils. n = 5 pooled mice. d. UMAP of Il1b response score (IL1r1, Il1r2, Il6, Tnf, Ccl2, Ccl3, Ccl4, Nfkbia, Nfkbiz, Ptgs2, IL1rn) across all aortic leukocyte populations. Representative flow cytometry plot showing IL-1R1 expression in aortic macrophages. e. Number of significant interactions of CD45.2 macrophages with wild type (CD45.1) macrophages in the aorta per condition, obtained from CellChat analyses. f. Significant ligand-receptor interactions between JAK2V617F and wild type macrophages in the aorta, grouped by lifestyle condition, obtained from CellChat analyses. g. Volcano plots showing differentially expressed genes in wild type (CD45.1) macrophages in Ldlr−/−Jak2V617FBM + SF (left panel) and Ldlr−/−Jak2V617FBM + exercise (right panel), compared with Ldlr−/−Jak2V617FBM mice. Extended Data Fig. 8 Contribution of neutrophils to the effects of sleep and exercise in Ldlr−/−Jak2V617FBM mice. a. UMAP and cluster proportions of reclustered aortic CD45.2 neutrophils across lifestyle groups in Ldlr−/−Jak2V617FBM mice. Volcano plots of DEGs comparing sedentary and WT, sedentary and SF, and sedentary and exercise CD45.2 neutrophils. b. NETosis score in CD45.2 neutrophils across lifestyle groups (Elane, Mpo, Ltf, Ngp, Camp, Padi4, H2afx, Hmgb1, Ncf1, Ncf2, Ncf4, Cybb, Gsdmd, Rac2, Plcg2, Fpr1, Tlr4, Cd177). n = 5 pooled mice. c. Representative flow cytometry plots of MPO and CitH3 positivity in CD45.1 WT and CD45.2 Jak2V617F neutrophils exposed to either no stimulation, PMA/Ionomycin, or LPS. Quantification of MPO+CitH3+ CD45.2 WT or Jak2V617F neutrophils exposed to either no stimulation, PMA/Ionomycin, or LPS across the 4 lifestyle groups (n = 3 wt, 4 sed, 4 SF and 4 exercise for all three interventions and both CD45.1 and CD45.2 neutrophils, respectively. d. Experimental schematic; exemplary flow cytometry plots showing neutrophil depletion in the blood in the anti-Ly6G group compared to IgG injected controls. Bodyweight, plasma cholesterol, and plasma triglyceride levels in Ldlr−/−Jak2V617FBM mice injected with IgG or anti-Ly6G across lifestyle groups at the time of euthanasia. (n = 7 sedentary + IgG, 8 sedentary + anti-Ly6G, 7 SF + IgG, 7 SF + anti-Ly6G, 4 exercise + IgG and 7 exercise + anti-Ly6G). e. Expansion of blood CD45.2 Jak2V617F monocytes in mice injected with IgG or anti-Ly6G across lifestyle groups relative to baseline (n = 3 sedentary + IgG, 8 sedentary + anti-Ly6G, 4 SF + IgG, 8 SF + anti-Ly6G, 4 exercise + IgG and 6 exercise + anti-Ly6G). Schematics in panels c–e created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). f. Lesion area, volume, and proportion of the necrotic core in Ldlr−/−Jak2V617FBM mice exposed to sedentary lifestyle, SF, exercise plus either IgG or anti-Ly6G with representative aortic root images stained with Oil-red-O (n = 6 sedentary + IgG, 8 sedentary + anti-Ly6G, 7 SF + IgG, 7 SF + anti-Ly6G, 4 exercise + IgG and 5-6 exercise + anti-Ly6G). Scale bars are 200 µm. Two-sided statistics. Data are mean ± s.e.m; *p < 0.05, **p < 0.01. P-values were obtained from individual Mann–Whitney U tests. Extended Data Fig. 9 Aortic inflammasome activation in Ldlr−/−Tet2−/−BM and Ldlr−/−p53−/−BM mice exposed to SF and extended analysis of CLEC4E signaling. Immunofluorescence staining of atherosclerotic plaques in Ldlr−/−Tet2−/−BM (a) and Ldlr−/−p53−/−BM (b) mice exposed to SF or sedentary lifestyle. Sections were stained for IL-1β, AIM2, and NLRP3, with Mac-2 as a counterstain to identify macrophages. Representative images are shown for each condition; scale bars are 400 µm. For Ldlr−/−Tet2−/−BM, n = 6, 6 (IL-1β) and 7, 7 (AIM2/NLRP3) for Sedentary vs SF. For Ldlr−/−p53−/−BM, n = 9, 9 (IL-1β, AIM2) and 8, 9 (NLRP3) for Sedentary vs SF. (c) UMAP of reclustered CD45.2 macrophages from single cell RNA sequencing of the aorta, integrating all cells across four conditions. Clec4e expression is visualized as a heat map overlaid per cell. Adjacent violin plot depicts Log2-transformed Clec4e expression across the four macrophage sub-clusters identified in the UMAP. n = 5 pooled mice. d. IL-1β in the supernatant of WT BMDMs with Clec4e agonism (TDB) or antagonism (n = 6 technical replicates control, 6 anti-CLEC4E, 6 anti-CLEC4E + TBD). e. Body weights and plasma lipids in Ldlr−/−Jak2V617FBM mice treated with either saline or anti-CLEC4E antibody and exposed to either SF or sedentary lifestyle at the time of euthanasia (n = 7, 7 sedentary + saline; 7,7 sedentary + anti-CLEC4E; 6, 6 SF + saline and 6, 5 SF + anti-CLEC4E for body weight and plasma lipids, respectively). Two-sided statistics. Data are mean ± s.e.m; ns = not significant, * p < 0.05, **** p < 0.0001. P-values were obtained from individual Mann–Whitney U tests (a) or ordinary one-way ANOVA (d). Schematic in panel e created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). Extended Data Fig. 10 Aortic inflammasome activation in Ldlr−/−Tet2−/−BM and Ldlr−/−p53−/−BM mice exposed to exercise and extended analysis of ADRβ2 signaling. Immunofluorescence staining of atherosclerotic plaques in Ldlr−/−Tet2−/−BM (a) and Ldlr−/−p53−/−BM (b) mice exposed to exercise or sedentary lifestyle. Sections were stained for IL-1β, AIM2, and NLRP3, with Mac-2 as a counterstain to identify macrophages. Representative images are shown for each condition. For Ldlr−/−Tet2−/−BM, n = 6, 6 (IL-1β) and 7, 6 (AIM2/NLRP3) for Sedentary vs exercise. For Ldlr−/−p53−/−BM, n = 9, 8 (IL-1β, AIM2) and 8, 8 (NLRP3) for Sedentary vs exercise c. UMAP of reclustered CD45.2 macrophages from single cell RNA sequencing of the aorta, integrating all cells across four conditions. Adrb2 expression is visualized as a heat map overlaid per cell. Adrb2 expression across the four CD45.2 aortic macrophage clusters in Ldlr−/−Jak2V617FBM mice. n = 5 pooled mice. d. Adrb2 expression in CD45.2 mutant and CD45.1 wild-type BM myeloid progenitors in Ldlr−/−Jak2V617FBM mice. e. Plasma norepinephrine levels in Ldlr−/− mice on a high cholesterol diet and exposed to lifestyle interventions (n = 6 sed, 6 SF, 5 exercise). f. Plasma corticosterone in Ldlr−/−WTBM and Ldlr−/−Jak2V617FBM mice exposed to lifestyle (n = 5 wt, 6 sed, 10 SF, 6 exercise). g. Adrb2 gene expression measured by qPCR in BMDMs from Ldlr−/−Jak2V617FBM mice subjected to exercise, expressed relative to values from sedentary Ldlr−/−Jak2V617FBM mice (n = 8). Body weight and plasma lipids (h), BM IL-1β (i), and immunofluorescence staining of atherosclerotic plaques (j) in Ldlr−/−Jak2V617FBM mice with or without exercise and exposed to saline or ICC-118,551. Sections were stained for NLRP3 and AIM2, with Mac-2 as a counterstain to identify macrophages. Representative images are shown for each condition. n = 8, 7-8 sedentary + saline; 5, 5 sedentary + ICC-118,551; 14, 10 exercise + saline and 8, 8 exercise + ICC-118,551 for body weight and plasma lipids in (h). n = 10, 5, 7, 8 and 10, 5, 10, 7 for identical groups in (i) and (j), respectively. All scale bars are 400 µm. Two-sided statistics. Data are mean ± s.e.m; * p < 0.05, ** p < 0.01, *** p < 0.001. P-values were obtained from independent, two-tailed, Welch’s t-tests (for parametric data) or Mann–Whitney U tests (for non-parametric data). Schematics in panels e,f,h created in BioRender; McAlpine, C. https://biorender.com/1rt7rau (2026). Supplementary information Source data Rights and permissions About this article Cite this article Gerhardt, T., Jacob, W., Gaebel, L. et al. Mutation-dependent responses to sleep and exercise in clonal haematopoiesis. Nature (2026). https://doi.org/10.1038/s41586-026-10634-0 Received: Accepted: Published: Version of record: DOI: https://doi.org/10.1038/s41586-026-10634-0