Caloric-restriction is necessary for efficacious weight loss, but can lead to unwanted losses of lean mass and bone mineral content. High-protein diets have been shown to protect lean mass during caloric-restriction and may have similarly preservative effects on bone either through attenuating increases in bone resorption or elevating bone formation. Objectives To determine whether increasing protein intake, in combination with daily aerobic exercise, attenuates elevations in bone resorption markers and reductions in bone formation markers during short-term severe caloric-restriction in healthy, active men. We hypothesized increasing protein intake would attenuate elevations in sclerostin and c-terminal telopeptide (CTX) while maintaining levels of procollagen type I propeptide (P1NP). Methods In this randomized crossover trial, participants underwent three conditions: caloric-restriction with low protein intake (CRLP; 0.8 g/kg body weight (BW)); caloric-restriction with high protein intake (CRHP; 1.7 g/kg BW) and energy balance control (CON; 1.7 g/kg BW). Caloric restriction was achieved by reducing energy availability, calculated as the difference between dietary energy intake and exercise energy expenditure, to 15 kcal/kg fat-free mass (FFM), whereas energy availability was maintained at 40 kcal/kg FFM in CON. Throughout all conditions, participants completed daily supervised exercise sessions on a bicycle ergometer set to expend 15 kcal/kg FFM. Before and after each five-day condition, body composition was assessed via bioimpedance analysis and markers of bone formation and resorption were assessed from fasting blood samples. Results Body weight decreased (p < 0.001) in CRLP [−1.97 ± 0.22 kg] and CRHP [−1.80 ± 0.52 kg], but remained stable in CON. Loss of fat mass was significant in CRHP [−1.14 ± 0.23 kg, p < 0.05] but not in CRLP [−0.57 ± 0.48 kg]. Sclerostin was elevated above baseline levels only in CRLP [19.7 ± 22.5 \%, p < 0.05] but not in CRHP [3.2 ± 18.3 \%] or CON [3.7 ± 26.1 \%]. CTX was reduced in CON [−8.9 ± 9.8 \%, p < 0.05] but did not change in CRHP [0.1 ± 5.4 \%] or CRLP [3.8 ± 14.7 \%]. P1NP did not change significantly in any group. Conclusions Elevating protein intake and performing daily aerobic exercise during five days of severe caloric restriction suppressed the elevation in sclerostin, a bone resorption marker, seen in individuals who performed daily aerobic exercise but did not elevate protein intake. This effect mirrors the protective effect of increased protein intake on lean mass. Support or Funding Information This work was supported in part by funding from the USDA National Institute of Food and Agriculture (NEB-36-083) and a Layman Seed Award from the University of Nebraska Foundation. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.