James Collins’ Wolves side and Mark Delaney’s Villa outfit are among the 16 youth teams who have been confirmed for the 2021/22 competition.
Wolves Under-21s made it out of their group last term – having club-record buy Fabio Silva score twice in a victory at Doncaster – before bowing out to Port Vale in the second round.
Villa were winless in their group, which also contained Fleetwood, Sunderland and Carlisle.
Albion, too, came at the bottom of their group and are not taking part this time around, with all the academy sides this year being from Premier League clubs.
Last season’s Trophy was eventually won by Sunderland, beating Tranmere 1-0 in the final at Wembley in March – after Salford won the Covid-delayed 2020 final 24 hours earlier.
The draw for round one of the Trophy will be made at 4.30pm Thursday and broadcast live on Sky Sports.