Background: We examined the impact of the Fuel Up to Play 60 (FUTP60) program on children's body mass index (BMI) and aerobic capacity (AC). Methods: Participation in the FUTP60 and both BMI and AC profiles were collected through the NFL PLAY 60 FitnessGram Partnership Project involving over 100 schools from 22 US states. We specifically examined the distributions of BMI and AC among participating versus nonparticipating schools in the 2012-2013 school year. Hierarchical linear models tested the impact of participation and availability of additional funding for program implementation on the proportions of youth meeting FitnessGram health-related fitness standards (i.e., Needs Improvement--Health Risk [NIHR] and Healthy Fitness Zone [HFZ] categories). Results: After 1 year implementing the program, participating schools had lower proportions of boys (-4.1 ± 2.0%, p = 0.04) and girls (-4.5 ± 2.0%, p = 0.03) in the NIHR for BMI, and lower proportion of girls (-9.7 ± 4.0%, p = 0.02) in the NIHR for AC. There were no differences in the distributions for the HFZ and the availability of additional funding did not alter the relationships (p > 0.05). Conclusions: This study provides preliminary evidence that participation in the FUTP60 is associated with improved profiles of health-related fitness.