PROJECT SUMMARY Mayo Clinic Chemical Center (MCC) is funded to perform targeted metabolites and exosome cargo measurements in samples collected in the Clinical Centers of MoTrPAC. As advised by the steering committee, we have expended a substantial amount of time developing an innovative methodology to isolate plasma exosomes and demonstrated reproducibility in multiple experiments, at different time points and by different laboratories. We propose exosome-based proteome and miRNA analysis in a limited number of samples. Based on power calculations, we estimated that the proposed number of samples will detect significant changes in exosome proteome and miRNA responses to aerobic and resistance exercise training. We will also test a hypothesis that proteome and miRNA release after acute exercise will be different in people who are highly active in comparison with sedentary people, because the prior exercise training accretes specific exercise-induced proteins in tissues that will be released during and immediately after both modes of exercise. These measurements will be done in 140 sedentary people and 110 highly active people and will likely offer unique new information. In addition, 55 sedentary people will be evaluated at baseline and following 3 months each of aerobic and resistance training with 30 sedentary controls studied after a 3-month control period. Secondly, in alignment with metabolomic measurements done at other centers, we will perform measurement of two quantitative targeted metabolites-amino metabolites and organic acids in plasma and skeletal muscle. These targeted metabolites are chosen based on their biological relevance to exercise training, and the quantitative and reproducible data anticipated from our assays would complement large scale metabolites measured in other Chemical Centers. The targeted metabolome measurement will also offer an opportunity to potentially understand the biochemical pathways involved in exercise benefits. We chose muscle and plasma measurements of organic acids representing the substrates of the citric acid flux pathway and amino metabolites which, based on many preliminary studies, are shown to sensitively respond to exercise with distinct responses to aerobic and resistance exercise. Both targeted metabolites in muscle and plasma as well as exosome-based measurements of proteome and miRNA are anticipated to reveal novel information on how specific exercise programs may benefit physiological functions. We anticipate that the results from our study in combination with the ongoing multiple omic measurements in all chemical centers will likely unravel deeper insights on how exercise benefits multiple organs. Our objective is to advance the translational potential of exosome cargo by administering biological molecules offering exercise benefits via small vesicles like exosomes to multiple organs in the body. The metabolites, especially targeted metabolites, are likely biomarkers to monitor responses to the administered vesicles carrying potentially beneficial molecules.

PROJECT NARRATIVE Here we propose measurements of molecules that are carried by small vesicles in the circulation delivering potential beneficial effects. We also are measuring quantitative metabolites which can offer evidence of exercise benefits and other therapeutic agents potentially carried by the small vesicles in the circulation.