Carbonic anhydrase XII mediates the survival and prometastatic functions of macrophages in human hepatocellular carcinoma
Wan-Ru Ning, … , Yan Wu, Limin Zheng
Wan-Ru Ning, … , Yan Wu, Limin Zheng
Published April 1, 2022
Citation Information: J Clin Invest. 2022;132(7):e153110. https://doi.org/10.1172/JCI153110.
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Carbonic anhydrase XII mediates the survival and prometastatic functions of macrophages in human hepatocellular carcinoma

Abstract

Macrophages constitute a major immune component in tumor tissues, but how these cells adapt to and survive in the nutrient-depleted and lactic acid–induced acidic tumor microenvironments is not yet fully understood. Here, we found that levels of carbonic anhydrase XII (CA12) expression were significantly and selectively upregulated on macrophages in human hepatocellular carcinoma (HCC). Transient glycolytic activation of peritumoral monocytes induced sustained expression of CA12 on tumor-infiltrating macrophages via autocrine cytokines and HIF1α pathways. On the one hand, CA12 mediated the survival of macrophages in relatively acidic tumor microenvironments, while on the other hand, it induced macrophage production of large amounts of C-C motif chemokine ligand 8 (CCL8), which enhanced cancer cell epithelial-mesenchymal transition (EMT) and facilitated tumor metastasis. Consistently, the accumulation of CA12+ macrophages in tumor tissues was associated with increased tumor metastatic potential and reduced survival of patients with HCC. Selective targeting of tumor-infiltrating macrophages with a CA12 inhibitor reduced tumor growth in mice and was sufficient to synergistically enhance the therapeutic efficacy of immune-checkpoint blockade. We suggest that CA12 activity is a previously unappreciated mechanism regulating the accumulation and functions of macrophages in tumor microenvironments and therefore represents a selective vulnerability that could be exploited in future designs for antitumor immunotherapeutic strategies.

Authors

Wan-Ru Ning, Da Jiang, Xing-Chen Liu, Yu-Fan Huang, Zhi-Peng Peng, Ze-Zhou Jiang, Tiebang Kang, Shi-Mei Zhuang, Yan Wu, Limin Zheng

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Figure 6

CCL8 released by tumor-associated monocytes and macrophages induces EMT and migration of hepatoma cells.

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CCL8 released by tumor-associated monocytes and macrophages induces EMT ...
(A–D) HepG2 cells were left untreated or treated with CCL8 (20 ng/ml) for 20 hours. Cell migration was analyzed (A and B) (n = 4). Expression levels of vimentin, E-cadherin, N-cadherin, SNAI1, SNAI2, TWIST1, and TWIST2 in the cells were determined by immunoblotting (C and D) (n = 5). One out of five representative graphs is shown in A. Scale bar: 200 μm. (E–G) CD14+ cells were purified from the peripheral blood of healthy donors. Cells were left untreated or treated with HepG2 TSN for 2 hours, washed, and cultured for another 48 hours before their supernatants were collected. HepG2 cells were then treated with CCM or TCM in the presence or absence of control IgG or anti-CCL8 neutralizing antibody (2 μg/ml) for 20 hours. HepG2 cell migration was analyzed (E and F) (n = 4), and the expression of vimentin, E-cadherin, and SNAI1 was determined by immunoblotting (G) (n = 5). One out of five representative graphs is shown in E. Scale bar: 200 μm. (H and I) CD14+ cells and cancer cells were purified from tumor tissues from 13 patients with HCC. Expression levels of CCL8 in CD14+ cells and VIM and CDH1 expression in cancer cells were determined by qPCR. Correlations between the mRNA levels of CCL8 and those of VIM or CDH1 were analyzed. Results shown in B and F are represented as mean ± SEM. P values were obtained by 2-tailed Student’s t test (B), 2-way ANOVA (F), or Pearson’s correlation and linear regression analysis (H and I). *P < 0.05.