Abstract

Prolonged antigen stimulation often induces CD8+ T-cell exhaustion, which undermines the efficacy of immunotherapy. To address this challenge, we developed Janus nanomotors (PML@fmPt NMs) constructed through cell membrane phase separation, providing an effective strategy for dynamic immunomodulation. In contrast to conventional nanoparticle fabrication, negatively charged metal nanoparticles induce spontaneous gel–fluid domain segregation on the membrane, generating a stable Janus structure with intrinsic asymmetry, modularity, and enhanced diffusional mobility. This bioinspired design enables dual inflammatory chemotaxis toward both the spleen and tumor tissues, achieving selective targeting of T cells and tumor cells. Nanomotors codelivered metformin and CRISPR/Cas9, synergistically reversing T-cell exhaustion and disrupting tumor tryptophan metabolism, a dual regulatory concept termed dual-output gear (DOG) therapy. In preclinical studies, the platform improved mitochondrial respiration of CD8+ T cells and significantly inhibited tumor growth. Beyond therapeutic efficacy, this work establishes a blueprint for intelligent biomaterial-based nanomotors, which highlights the potential of phase-separation engineering for fabricating next-generation nanomaterials: the modular Janus configuration enables facile customization, the use of natural membranes enhances compatibility, and the phase separation principle can be broadly extended to other delivery systems.