Fig. 1

Study design for linking physical activity (PA), body weight measures, plasma biomarkers, and the gut microbiome in the Men’s Lifestyle Validation Study (MLVS) and associations with overall gut microbiome configuration. a To associate the gut microbiome with PA and body weight measures, we profiled stool metagenomes, metatranscriptomes, and blood biomarkers from the MLVS. The MLVS is a sub-study of the Health Professionals Follow-up Study (HPFS), an ongoing prospective cohort of 51,529 men. The HPFS has repeatedly collected PA and body weight information using validated questionnaires and health-related information since 1986. In the period 2012–2013, the MLVS collected stool samples at up to four time points per individual, blood samples at up to two time points, and additional PA information using accelerometer and body weight information using doubly labeled water from 307 participants. We applied MetaPhlAn 2 and HUMAnN 2 to perform taxonomic and functional profiling from stool shotgun metagenomes and metatranscriptomes. Plasma biomarkers of inflammation (high-sensitivity C-reactive protein, CRP) and glucose homeostasis (hemoglobin A1c, HbA1c) were measured using standard methods. We employed generalized linear mixed-effects regression models to account for within-subject correlation due to repeated sampling and occasional missing data. b Discovery cohort, validation cohort, and main exposure and outcome variables used in this study. c Mean BMI at different ages and the time of stool collection. The error bars represent standard deviations. d Spearman correlation between main exposure and outcome variables. e Principal coordinate analysis of all samples using species-level Bray–Curtis dissimilarity. f Proportion of variation in taxonomy explained by PA measures, body weight measures, plasma biomarkers, and covariables as quantified by two-sided permutational multivariate analysis of variance (based on species-level Bray–Curtis dissimilarity)