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 3

Loss of KCs preferentially in the tumor core and periphery at the late stage of liver metastasis.

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Loss of KCs preferentially in the tumor core and periphery at the late s...
(A) Clec4f-iDTR and WT C57BL/6 mice were treated with DT prior to or (B) after B16F10 tumor cell inoculation, and their livers were harvested at the indicated time points. n = 5 mice per group. Mice with intrasplenically injected B16F10-ZsGreen cells were imaged at (C) day 1, (D) day 3, or (E) day 7 after injection. Scale bars: 40 (C); 20 (D); 100 μm (E). Original magnification, zoomed images: ×4.3 (C); ×3.8 (D); ×4.3 (E). Tumors that closely interacted with at least 3 KCs were considered KC-associated tumors, and their ratio was quantified at day 3 (D: right panel) and (F) day 7. (G) Localization of KCs with tumors of different sizes. KC dark zones are outlined between dashed lines. Scale bars: 140 μm. Original magnification, zoomed images: ×3.0. (H) Quantification of the area of the KC dark zone in G. (I) Quantification of the density of KCs in tumor-adjacent (0–200 μm away from the tumor edge) or distant areas (600–800 μm away from the tumor edge). For H and I, each circle represents 1 tumor. n = 12. (J) Representative flow cytometric plot of F4/80+TIM4+ resident KCs in tumor-free or tumor-bearing mice. (K) KC numbers per mouse are shown. n = 5 mice. Representative or pooled data from at least 2 independent experiments are shown. Data are represented as mean ± SEM. **P < 0.01; ***P < 0.001, unpaired Student’s t test (A, B and K); 1-way ANOVA with Tukey’s test in (H and I).