Elucidation of Spatial Cooperativity in Chemo‐Immunotherapy by a Sequential Dual‐pH‐Responsive Drug Delivery System

Abstract Combining immune checkpoint blockade with chemotherapy through nanotechnology is promising in terms of safety and efficacy. However, the distinct subcellular distribution of each ingredient's action site makes it challenging to acquire an optimal synergism. Herein, a dual‐pH responsive hybrid polymeric micelle system, HNP(αPDL1_6.9, Dox_5.3), is constructed as a proof‐of‐concept for the spatial cooperativity in chemo‐immunotherapy. HNP retains the inherent pH‐transition of each polymer, with stepwise disassembly under discrete pH thresholds. Within weakly acidic extracellular tumor environment, αPDL1 is first released to block the checkpoint on cell membranes. The remaining intact Doxorubicin‐loaded micelle NP(Dox)_5.3 displays significant tropism toward tumor cells and releases Dox upon lysosomal pH for efficient tumor immunogenic cell death without immune toxicity. This sequential‐released pattern boosts DC activation and primes CD8⁺ T cells, leading to enhanced therapeutic performance than single agent or an inverse‐ordered combination in multiple murine tumor models. Using HNP, the indispensable role of conventional type 1 DC (cDC1) is identified in chemo‐immunotherapy. A co‐signature of cDC1 and CD8 correlates with cancer patient survival after neoadjuvant Pembrolizumab plus chemotherapy in clinic. This study highlights spatial cooperativity of chemo‐ and immuno‐agents in immunoregulation and provides insights into the rational design of drug combination for future nanotherapeutics development.