Phase-Separation Engineered Nanomotors Enable Spleen-Tumor Dual Targeting to Reverse T-Cell Exhaustion.

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.