Tottenham midfielder will miss the Champions League tie with Manchester City
Winks has a groin injury and has been left at home for treatment, the club confirmed.
Eric Dier and Serge Aurier have also been ruled out.
Winks started Tottenham's win over City last week, in which the North London side registered a 1-0 first half lead.
Tottenham are now expected to start with Moussa Sissoko and Victor Wanyama in central midfield.
Winks' injury is a problem for Tottenham beyond tonight. Spurs are battling for a top four place and cannot afford any more fitness issues.
Winks suffered with injuries last season and had been largely fitness free this season until this blow, at the worst possible time.
Here is a look at how Tottenham fans reacted to Winks' blow...
Winks not being available tomorrow is a massive blow for us.— Altug Kop WBFF Pro (@AltuKop) April 16, 2019
Never thought I would have said that last season, but he's been brilliant. We'll miss his composure on the ball. #COYS
looks like it’s Wanyama & Sissoko as the two holding midfielders tomorrow then - missing Harry Winks & Harry Kane is a massive massive blow! Absolutely vital we don’t concede in first 20mins to stand any chance! #COYS #THFC #MCITOT— Paul Redshaw (@PReddog84) April 16, 2019
No Winks is a huge blow. Our injuries this season have decimated the team, no idea how we are where we are. One final push #coys— Jamie Leigh (@JLTHFC1) April 16, 2019