Winger Chris Ashton has been ruled out of England’s Six Nations match against Wales on Saturday with a calf strain.
The Sale Sharks back made his first Six Nations start since 2013 in the win over France but has been left out of coach Eddie Jones’ 33-man squad.
On Tuesday, England prop Mako Vunipola was ruled out of the championship with an ankle injury while lock Maro Itoje continues his rehab with a knee issue.
England play Wales in Cardiff on Saturday, 23 February, at 16:45 GMT.
Both sides won their first two games in this year’s Six Nations with England top of the table ahead of second-placed Wales thanks to two bonus points.
Exeter Chiefs winger Jack Nowell started England’s opening win over Ireland and could return in Ashton’s place, while Bath’s Joe Cokanasiga is also in the squad.
The RFU says Ashton has a “minor calf strain” and is expected to recover in time to play Italy in the penultimate round on 9 March.
Saracens lock Itoje, meanwhile, is considered highly unlikely to play against Wales, but has not yet been ruled out by the England management.
Hooker Dylan Hartley, who definitely will not be available, has returned to his club Northampton to continue his recovery from a knee problem.
England are due to name their matchday squad on Thursday.
England squad in full
Forwards: Cole (Leicester Tigers), Cowan-Dickie (Exeter Chiefs), T Curry (Sale Sharks), Earl (Saracens), Ewels (Bath Rugby), Genge (Leicester Tigers), George (Saracens), Hepburn (Exeter Chiefs), Hughes (Wasps), Kruis (Saracens), Launchbury (Wasps), Lawes (Northampton Saints), Moon (Exeter Chiefs), Shields (Wasps), Sinckler (Harlequins), Singleton (Worcester Warriors), B Vunipola (Saracens), Harry Williams (Exeter Chiefs), Mark Wilson (Newcastle Falcons)
Backs: Brown (Harlequins), Cokanasiga (Bath Rugby), Daly (Wasps), Devoto (Exeter Chiefs), Farrell (Saracens, captain), Ford (Leicester Tigers), May (Leicester Tigers), Nowell (Exeter Chiefs), Robson (Wasps), Slade (Exeter Chiefs), Te’o (Worcester Warriors), Thorley (Gloucester Rugby), Tuilagi (Leicester Tigers), Youngs (Leicester Tigers)