Advances in nanotechnology have brought innovations to cancer therapy. Nanoparticlebased

anticancer drugs have achieved great success from bench to bedside. However, insufficient

therapy efficacy due to various physiological barriers in the body remains a key challenge. To

overcome these biological barriers and improve the therapeutic efficacy of cancers, multistage selfassembled nanomaterials with advantages of stimuli-responsiveness, programmable delivery, and immune modulations provide great opportunities. In this review, we describe the typical biological barriers for nanomedicines, discuss the recent achievements of multistage self-assembled nanomaterials for stimuli-responsive drug delivery, highlighting the programmable delivery nanomaterials, in situ transformable self-assembled nanomaterials, and immune-reprogramming nanomaterials. Ultimately, we perspective the future opportunities and challenges of multistage self-assembled nanomaterials for cancer immunotherapy.