This is not the first time the couple has separated and filed for divorce. They have two children together.
Robert De Niro and his wife, Grace Hightower, have reportedly called it quits after over 20 years of marriage, according to multiple outlets.
A source close to the former couple told Page Six on Tuesday that De Niro, 75, and Hightower, 63, are "not living together at this time."
“Sometimes things don’t work out the way you hope or want them to,” another insider told People magazine.
A rep for De Niro did not immediately respond to Fox News' request for comment.