Increasing heat in urban environments has recently become one of the most dangerous climate hazards due to its adverse impacts on urban populations. Implementing street-level trees could be an effective strategy to mitigate pedestrian heat exposure, particularly due to their ability to block incoming solar radiation. In this study, the PALM model system was applied to simulate the effects of a tree canopy and its location on heat exposure, as quantified by the Universal Thermal Climate Index (UTCI), during a heat wave, using the example of Prague-Dejvice, Czech Republic. Our results show that trees reduce the UTCI under their canopy by 3.5 °C on average, with the greatest UTCI reduction in open spaces during mornings and afternoons. High spatio-temporal variations in the reduction of UTCI by a tree canopy were observed in the study domain, especially in street canyons and courtyards. The effectiveness of trees in mitigating heat exposure was found to be closely related to their individual location with respect to surrounding buildings, specifically: (i) the distance from the nearest building, (ii) the height of the nearest building, and (iii) the azimuthal angle of the vector from the nearest building towards the tree. Model simulations indicate that a particularly small reduction in UTCI (about 2.5 °C less than the mean) can be found under trees located in the shade of taller buildings that are within a few metres and between southwest and southeast of the trees. Our findings illustrate that tree positioning in cities should be undertaken carefully and thoughtfully so that the presence of trees effectively improves thermal comfort and urban quality of life.