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Transcription factor Zhx2 is a checkpoint that programs macrophage polarization and antitumor response

Cell Death & Differentiation volume 30, pages 2104–2119 (2023)Cite this article

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Abstract

Macrophages are usually educated to tumor-associated macrophages (TAMs) in cancer with pro-tumor functions by tumor microenvironment (TME) and TAM reprogramming has been proposed as a potential tumor immunotherapy strategy. We recently demonstrated the critical role of Zinc-fingers and homeoboxes 2 (Zhx2) in macrophages’ metabolic programming. However, whether Zhx2 is responsible for macrophage polarization and TAMs reprogramming is largely unknown. Here, we show that Zhx2 controls macrophage polarization under the inflammatory stimulus and TME. Myeloid-specific deletion of Zhx2 suppresses LPS-induced proinflammatory polarization but promotes IL-4 and TME-induced anti-inflammatory and pro-tumoral phenotypes in murine liver tumor models. Factors in TME, especially lactate, markedly decrease the expression of Zhx2 in TAMs, leading to the switch of TAMs to pro-tumor phenotype and consequent cancer progression. Notably, reduced ZHX2 expression in TAM correlates with poor survival of HCC patients. Mechanistic studies reveal that Zhx2 associates with NF-κB p65 and binds to the Irf1 promoter, leading to transcriptional activation of Irf1 in macrophages. Zhx2 functions in maintaining macrophage polarization by regulating Irf1 transcription, which may be a potential target for macrophage-based cancer immunotherapy.

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Fig. 1: Reduced expression of Zhx2 in tumor associated macrophages.
Fig. 2: Lactate suppressed Zhx2 expression in TAM.
Fig. 3: Deficiency of Zhx2 in macrophages aggravated tumor burden.
Fig. 4: Zhx2-deficient macrophages shared an M2-like phenotype and promoted suppressive T cell infiltration in TME.
Fig. 5: Zhx2 promoted macrophages towards the M1 phenotype.
Fig. 6: Zhx2 directly promotes Irf1 transcription in macrophages and subsequent inflammatory responses.
Fig. 7: NF-κB P65 is required for Zhx2 mediated enhancement of Irf1 transcription and macrophage M1 polarization.

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Data availability

The RNA sequencing raw data have been deposited in NCBI Sequence Read Archive (SRA) database with the accession number: PRJNA598552. Uncropped original western blots and relevant data are provided in the Supplementary files. The data generated in this study are available upon reasonable request from the corresponding author.

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Acknowledgements

This study was supported in part by the National Key Research and Development Program (2021YFC2300603), the National Science Foundation of China (Key program 81830017, 8223056, 32200742), Taishan Scholarship (No.tspd20181201), Major Basic Research Project of Shandong Natural Science Foundation (No. ZR2020ZD12), the Shandong Provincial Natural Science Foundation (ZR2021QH322, ZR2019PH023) and Collaborative Innovation Centre of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong. We also thank the Translational Medicine Core Facility of Shandong University for the consultation and instrument availability that supported this work.

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  1. These authors contributed equally: Siyu Tan, Zehua Wang.

Authors and Affiliations

  1. Department of General Surgery, Qilu Hospital, Shandong University, Jinan, China

    Siyu Tan & Tao Li

  2. Key Laboratory for Experimental Teratology of Ministry of Education, Key Laboratory of Infection and Immunity of Shandong Province and Department of Immunology, School of Basic Medical Sciences, Qilu Hospital, Cheeloo Medical College of Shandong University, Jinan, Shandong, China

    Siyu Tan, Zehua Wang, Na Li, Xiaowei Guo, Yankun Zhang, Hongxin Ma, Xueqi Peng, Ying Zhao, Lifen Gao, Xiaohong Liang & Chunhong Ma

  3. Department of Clinical Laboratory, Qilu Hospital, Shandong University (Qingdao), Qingdao, China

    Zehua Wang

  4. Department of Clinical Laboratory, Shandong Cancer Hospital and Institute, Shandong First Medical University, and Shandong Academy of Medical Sciences, Jinan, China

    Hongxin Ma

  5. Key Laboratory for Experimental Teratology of Ministry of Education, Department of Histology and Embryology, School of Basic Medical Sciences, Cheeloo Medical College of Shandong University, Jinan, Shandong, China

    Chunyang Li

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Contributions

CM, ST, and ZW formulated the study concept and designed the studies. ST, ZW, NL, XG, YZ, HM, XP, and YZ performed the experiments. ST, ZW, and CM analyzed the results. CL, LG, TL, XL, and CM interpreted the results. ST, ZW, and CM wrote and edited the manuscript.

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Correspondence to Xiaohong Liang or Chunhong Ma.

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Tan, S., Wang, Z., Li, N. et al. Transcription factor Zhx2 is a checkpoint that programs macrophage polarization and antitumor response. Cell Death Differ 30, 2104–2119 (2023). https://doi.org/10.1038/s41418-023-01202-4

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