In Marvel Villainous: Twisted Ambitions, players can forge their own twisted paths to victory as Doctor Octopus, Titania, and Kang the Conqueror. To win as Doctor Octopus, players must complete five schemes, such as defeating Spider-Man or amassing and paying power. Those who pick Titania will need to gain strength to become empowered and defeat She-Hulk. Finally, Kang the Conqueror will require players to unleash variants of Kang into other villains' decks and conquer four locations in other villain domains with robot duplicates.

Each of the characters in Marvel Villainous: Twisted Ambitions can be played on their own or be mixed and matched with villains from previous Marvel Villainous releases.