First-team players Tony Clubb and Jake Shorrocks are facing long spells on the sidelines.
Clubb, 29, needs an operation which is expected to keep him out of action until later this campaign.
He missed the Easter weekend games because of what was described as a “lower back problem”.
Shorrocks, 21, is unexpected to play again in 2017 after suffering a knee injury in training.
Rugby manager Kris Radlinski explained: “Tony has a medical condition, unrelated to any injury, that he has been managing for many years.
“Having struggled with it recently doctors have advised that he will need a routine operation.
“Tony is expected to make a full recovery but will need time to recover from the surgery. It’s difficult to put a timeframe on a return but we are hopeful that he might play again later this season.
“Jake picked up an injury in the Widnes game and recovered well from that only to suffer a severe knee injury in non-contact training. We are unable to put a timeframe on the injury however we don’t expect him to play again this season.
“It’s a tough pill to swallow for both Tony and Jake but we will give them all of our support, as I’m sure will the Wigan fans, and we look forward to seeing them back out on the field playing for Wigan.”
Clubb, who has featured in nine games for Wigan this season, missed most of last season with a serious neck injury.
Shorrocks, a half-back who has also covered at hooker, has played just once for Wigan this year in the Super League game against Widnes.
Shaun Wane has four other senior players on the sidelines – Dom Manfredi, Sam Tomkins, Oliver Gildart and John Bateman.