In situ expansion and reprogramming of Kupffer cells elicit potent tumoricidal immunity against liver metastasis
Wei Liu, … , Lu Li, Zhutian Zeng
Wei Liu, … , Lu Li, Zhutian Zeng
Published February 23, 2023
Citation Information: J Clin Invest. 2023;133(8):e157937. https://doi.org/10.1172/JCI157937.
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Research Article Hepatology Immunology

In situ expansion and reprogramming of Kupffer cells elicit potent tumoricidal immunity against liver metastasis

Abstract

Liver metastasis represents one of the most frequent malignant diseases with no effective treatment. Functional reprogramming of Kupffer cells (KCs), the largest population of hepatic macrophages, holds promise for treating liver cancer, but remains seldom exploited. Taking advantage of the superior capacity of KCs to capture circulating bacteria, we report that a single administration of attenuated Escherichia coli producing clustered regularly interspersed short palindromic repeats CasΦ (CRISPR/CasΦ) machinery enables efficient editing of genes of interest in KCs. Using intravital microscopy, we observed a failure of tumor control by KCs at the late stage of liver metastasis due to KC loss preferentially in the tumor core and periphery, resulting in inaccessibility of these highly phagocytic macrophages to cancer cells. Simultaneous disruption of MafB and c-Maf expression using the aforementioned engineered bacteria could overcome KC dysfunction and elicit remarkable curative effects against several types of metastatic liver cancer in mice. Mechanistically, bacterial treatment induced massive proliferation and functional reprogramming of KCs. These cells infiltrated into the tumor, dismantled macrometastases by nibbling cancer cells, and skewed toward proinflammatory macrophages to unleash antitumor T cell responses. These findings provide an immunotherapy strategy that could be applicable for treating liver metastasis and highlight the therapeutic potential of targeting tissue-resident macrophages in cancer.

Authors

Wei Liu, Xia Zhou, Qi Yao, Chen Chen, Qing Zhang, Keshuo Ding, Lu Li, Zhutian Zeng

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Figure 6

KC-dependent elimination of liver metastasis during BIL-CRISPR–mediated bacterial therapy.

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KC-dependent elimination of liver metastasis during BIL-CRISPR–mediated ...
(A) Mice with established B16F10 liver metastasis were treated with E. coli–vector or E. coli–sgMafb/Maf i.v. Representative intravital liver images at 7 days after bacterial treatment are shown. Scale bars: 100 μm. Original magnification, zoomed images: × 3.8. Peritumoral KC dark zones are show between yellow and blue dashed lines. (B) Intravital liver images of tumor-KC interfaces at various time points after bacterial treatment. Scale bars: 100 μm. Original magnification, zoomed images: × 2.9. (C) Representative time-lapse 3D intravital liver images showing KCs nibbling an interacting cancer cell after bacterial therapy. Scale bars: 40 μm (red); 15 μm (black). (D) WT or Clec4f-iDTR mice with established B16F10 liver tumors were treated with DT 2 days after ClearColi sgMafb/Maf treatment, and the livers were harvested 5 days later, as depicted. Scale bars: 1 cm. (E) Liver weights and tumor area on the surface of livers were measured in D. n = 9–10 mice per group pooled from 3 independent experiments. (F) Ccr2–/– mice with established B16F10 liver tumors were injected with ClearColi-vector or ClearColi-sgMafb/Maf at day 12 and harvested at day 19. (G and H) Liver weights and the tumor area on the surface of livers from F were measured. n = 4 mice from 1 of 2 independent experiments. Data are represented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001, unpaired Student’s t test.