He began his career in motorsport in karting in 1999. By 2002, he was a three-time Belgian champion as well as winning the acclaimed Junior Monaco Kart Cup in 2000. He also won the World Cup Formula A championship in 2002.
D’Ambrosio graduated to single-seater racing in 2003, winning the Belgian Formula Renault championship at the first attempt and then competing in Formula Renault 3.5, Euro 3000 series, Formula Masters and GP2. In 2009 he was runner-up in GP2 Asia and won a race in the main series securing a place as a test driver for the Virgin Racing Formula 1 team.
In Formula 1, Jerome raced for Marussia Virgin Racing and Lotus F1 Team before moving to Formula E where he competed until 2020. In October that year d’Ambrosio joined the Venturi Racing Formula E team as Deputy Team Principal and then as Team Principal, guiding the team to ten podiums in 16 races and second place in the championship.
In 2023 d'Ambrosio joined the Mercedes F1 Team working closely with Team Principal Toto Wolff, as well as acting as the Driver Development Director for the team.
In July 2024 it was announced that Jerome was joining Scuderia Ferrari HP as Deputy Team Principal and Head of Scuderia Ferrari Driver Academy as from October 1.