Changes in tidal regime in the heavily modified Venice Lagoon, Italy, are investigated using long-term observations and numerical modelling. The amplitudes of the major tidal constituents exhibit a significant increase over the last century. Analysis of tide gauge data in the adjacent Adriatic Sea reveals that these changes could be only partially attributed to the rise of the mean sea level. Numerical experiments confirm that natural and anthropogenic morphological changes are responsible for the alteration of tidal regime inside the lagoon. Temporal and spatial changes in tidal asymmetry highlight the complex impacts of human interventions on tidal changes and long-term morphodynamics. Our results suggest that over time the lagoon became more and more an ebb-dominant system. Moreover, in Venice the tidal modulations are significantly impacting the frequency with which high water level thresholds are exceeded. Occurrence of flooding events is therefore influenced by sea level rise and secondarily by the increase in amplitude of principal tidal waves.