Miles Morales is a teenager who struggles to adjust to his new elite boarding school and live up to the expectations of his parents, nurse Rio Morales and cop Jefferson Davis, who see Spider-Man as a menace, despite Miguel, who see him as a hero. After developing a crush on his classmate Gwanda, Miles takes advantage his uncle Aaron Davis, who encourages Miles to pursue his passion for graffiti and leads him to a subway station where he can draw. Once there, Miles is bitten by a radioactive spider and develops spider-like abilities. Unable to contact Aaron, he returns to the station and finds a secret laboratory where Wilson Fisk has built a particle accelerator to access parallel universes. Spider-Man arrives to disable the accelerator fearing that the accelerator can to destroy the city and fights Green Goblin and Prowler. During the fight, Spider-Man is touched by the accelerator ray, seeing the multiverse, and he is hurt, falling down. Spider-Man gives Miles a pen-drive to disable the accelerator and reports him that the machine could destroy the city if it's turned on again. Shortly afterward, Fisk kills brutally Spider-Man as Miles escapes. Alone and without help, Miles visits Parker's tomb trying to know if he will be capable to make the job when another Peter Parker appears behind him. Running from the cop, Miles learns that this new Peter Parker he meets belongs to another dimension from where he was abducted during the experiment. Looking for make this else-Parker his trainer, they try to steal the accelerator computer to create a new pen-drive after to know that the previous was accidentally broken. But in the assault to the laboratory they realize that the scientist who lead the project is a Dr. Octopus' female-version. Escaping from the lab, else-Parker and Miles are helped by Gwanda, in true a Gwen Stacy from another dimension who turned in Spider-Gwen after she wasn't prevent the death of the Peter Parker of her dimension. Without options, the three visit aunt May, just to learn that another three alternate counterparts more await them: Spider-Man Noir, a black-and-white 30's years stylized Spider-Man who fights against Nazis in 1937; Penny Parker, a 14 years old Anime-version Japanese schoolgirl from the year 3145 with a powerful Spider-Robot; and Spider-Ham, a funny cartoon-version named Peter Porker. But things complicate for Miles when he discovers that his uncle Aaron works as Kingpin's henchman Prowler, being assassinated by the own Kingpin when Aaron couldn't kill Miles after to learn Miles as new Spider-Man. With his family broken and unable to control his Spider-powers, Miles tries assume his place in the world and the responsibility as the new Spider-Man, at the same time that Kingpin reconnects the accelerator obsessed to recover an alternate version of his wife and son, who died in a car accident when they ran from him while he fought against Spider-Man. Realizing that all his five counterparts are in danger to die disintegrated for being in an universe that it isn't their own, Miles faces his destiny to save not only all them partners, but the universe from Kingpin's ambitions.