Lucas Tousart scored the only goal in the first half as the hosts claimed their first victory over the Italian giants.
Juve commenced confidently however failed to create any clear chances earlier than Tousart converted Houssem Aouar’s cross from close range to hand the Ligue 1 side the lead.
The visitors persisted to have extra possession but they struggled to penetrate the resolute Lyon defence, failing to register a single shot on target.
With the clock ticking down, Juve ramped up the pressure, however Paulo Dybala’s late strike was ruled out for offside.
The 2nd leg will take place at Allianz Stadium on 17 March.