Spurs welcome the seven-time French champions to White Hart Lane for the first leg of their last-32 tie.
"It's a difficult tie against a team that is very, very experienced and have many trophies in the bag," Villas-Boas told Tottenham's official website. "They also have a lot of experience in the Champions League.
"Our players are motivated to do well in this competition and they know that if they beat Lyon, it will serve as further inspiration for the rest of the tournament."
Both sides failed to lose a game during their group campaigns in the competition.