The film contains no identifiable LGBTQ+ characters or themes. Interactions with alien entities include neutral pronouns and fan-interpreted queerplatonic elements, but these do not constitute explicit queer human representation or narrative focus.

The film contains no identifiable transgender characters or themes. Its narrative focuses on interstellar survival and interspecies friendship without addressing gender identity.

Female characters, including Eva Stratt played by Sandra Hüller, hold authoritative roles in the project's leadership but do not participate in physical combat. The narrative emphasizes scientific challenges, space exploration, and interspecies cooperation without depictions of hand-to-hand fights or melee confrontations.

The film's main characters, including Ryland Grace and Eva Stratt, retain their canonical genders from Andy Weir's novel, with no reported instances of gender swaps in the adaptation.

The 2026 adaptation of Project Hail Mary retains the racial portrayals of its human characters as established in Andy Weir's novel, with international crew members cast to match their nationalities and no alterations to established ethnicities for legacy roles.