Aquifer thermal energy storage systems may support the decarbonization of heating and cooling energy needs of urban areas, not only in heating-dominated countries but also in Southern Europe. In this framework, this work investigates the adoption of a water-to-water heat pump interacting with two aquifers and activating a small-scale district heating and cooling network serving a mixed-use district of eight residential and office buildings in Rome (Italy). The dynamic behaviour of aquifers has been replicated using the GeoSIAM software. Energy conversion systems and users’ thermal and cooling loads have been simulated in TRNSYS 18. The dynamic models have been integrated using an iterative approach based on conditions regarding plant operation and injection temperature in wells. The proposed solution has been then compared from the energy and environmental perspective with a traditional system, including an air-to-water heat pump. In addition, the balance between heating and cooling mode operation has been assessed. The results obtained encourage the adoption of aquifer thermal energy storage systems in Central Italy. Indeed, the primary energy saving, and the carbon dioxide emissions avoided are equal to 18%, whereas the imbalance between cooling and heating loads is limited to -5.2%.