PURPOSE: To implement, validate, and apply a self-gated free-running whole-heart five-dimensional (5D) flow MRI framework to evaluate respiration-driven effects on three-dimensional (3D) hemodynamics in a clinical setting. MATERIALS AND METHODS: In this prospective study, a free-running five-dimensional (5D) flow sequence was implemented with 3D radial sampling, self-gating, and a compressed-sensing reconstruction. The 5D flow was evaluated in a pulsatile phantom and adult participants with aortic and/or valvular disease who were enrolled between May and August 2019. Conventional twofold-accelerated four-dimensional (4D) flow of the thoracic aorta with navigator gating was performed as a reference comparison. Continuous parameters were evaluated for parameter normality and were compared between conventional 4D flow and 5D flow using a signed-rank or two-tailed paired t test. Differences between respiratory states were evaluated using a repeated-measure analysis of variance or a nonparametric Friedman test. RESULTS: A total of 20 adult participants (mean age, 49 years ± 17 [standard deviation]; 18 men and two women) were included. In vitro 5D flow results showed excellent agreement with conventional 4D flow–derived values (peak and net flow