Taking a hydrogen-fueled truck frame as the research object, based on the topology optimization theory of variable density method, the goal of maximizing the static multi-working stiffness and improving the dynamic low-order natural frequency of the frame is realized. The attachments of the frame were retained, and the topological optimization space was established according to the original frame outline size. A multi-objective topology optimization was carried out by using the isotropic material penalty function to obtain the best frame structure layout, and then the new frame structure was designed. The results show that the middle bearing capacity of the new frame is better than that of the original frame which has been verified by modal testing, and the maximum stress decreases by 35.6%. The first-order natural frequency increases by 1.9 Hz, which can avoid the maximum excitation energy region of the road. Meanwhile, the frame weight is reduced by 7.8%, which realizes the specialized and lightweight design of a hydrogen-fueled truck frame.