As a four-year starter at the University of Miami, Wayne set a school record of 173 career catches (including 36 consecutive games with a reception) and is one of only four wide receivers in school history to post 20 or more touchdowns in his career, along with Michael Irvin, Lamar Thomas, and Leonard Hankerson. His 48 receptions as a freshman were a school record. Wayne graduated with a degree in liberal arts. He was also roommate to Baltimore Ravens safety Ed Reed.
Wayne was inducted into the University of Miami Sports Hall of Fame at their 43rd Annual Induction Banquet held on March 24, 2011.
In Wayne's first year, he caught 27 passes for 345 yards with no touchdowns. His yards doubled the next year as he caught 49 passes for 716 yards and four touchdowns.
Wayne broke into the starting lineup for good in the 2003 season, and he caught 68 balls for 838 yards and seven touchdowns. The next season, Wayne caught 77 passes for 1,210 yards and 12 touchdowns during a season in which Colts quarterback Peyton Manning set a then-record by throwing for 49 touchdowns. During the playoffs, Wayne was named the "Wild Card Weekend Offensive Player of the Week" after the Colts beat the Broncos. In 2005 Wayne continued to increase his yearly reception total with 83.
In 2006, Wayne signed a 6-year, $39.5 million contract. He went on to catch 86 balls for a then career high 1,310 yards and 9 touchdowns. As a result, Wayne was selected to his first Pro Bowl. Wayne helped the Colts defeat the Chicago Bears in Super Bowl XLI with a 53-yard touchdown reception in the first quarter.
In 2007, Wayne responded to a rash of injuries on the team, including Harrison and tight end Dallas Clark, and set career highs in receptions (104), and yards (1510). He led the league in receiving yards and was selected to go to the Pro Bowl for the second consecutive year.
In 2008, Wayne once again posted another Pro Bowl season with 82 catches for 1145 yards and six touchdowns, including a 65-yard score in the opening drive against the Pittsburgh Steelers.
In 2009, Wayne caught the winning touchdown pass against the New England Patriots in what is now know as the "4th and 2" game. Wayne was also selected as a starter for the Pro Bowl, but couldn't play due to the Colts playing in the Super Bowl.
In 2010, Wayne was second in the NFL receptions with 111, and third in receiving yards with 1,355. During a season where the Colts heavily relied on Peyton Manning to win games, Wayne again cemented himself as one of the top receivers in the NFL. Wayne was also selected as a starter the Pro Bowl.
On March 13, 2012, the Colts re-signed Wayne to a three-year contract.