Engineered probiotics for tumor-targeted combination chemoimmunotherapy

Abstract Achieving tumor-specific delivery and sustained activation of both cytotoxic and immune-modulating agents remains a critical challenge in chemoimmunotherapy. Here, we present a bacterial platform engineered to combine enzyme/prodrug chemotherapy with immunotherapy, where tumor-homing E. coli Nissle 1917 expresses cytosine deaminase to convert the prodrug 5-fluorocytosine into the cytotoxic drug 5-fluorouracil within tumors. Concurrently, the engineered bacteria produce an IL-15 superagonist and a PD-L1 blocking nanobody to mitigate the immunosuppressive effects of tumor-localized chemotherapy. This platform demonstrated potent antitumor effects in the murine MC38 solid tumor model. Mechanistic studies showed that the combination therapy enhances activation of antigen-presenting cells, T cells and natural killer cells, while reducing immunosuppressive populations. In summary, our approach integrates enzyme/prodrug therapy and immunotherapy into a single bacterial delivery system, overcoming the limitations of conventional therapies and offering a scalable and precision-engineered strategy with an improved safety profile for synergistic cancer treatment. One Sentence Summary A precision-engineered bacterial platform integrates enzyme/prodrug chemotherapy and immunotherapy to drive synergistic antitumor responses with enhanced safety, offering promise for clinical translation.