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Phospholipid peroxidation in macrophage confers tumor resistance by suppressing phagocytic capability towards ferroptotic cells

Cell Death & Differentiation volume 31, pages 1184–1201 (2024)Cite this article

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Abstract

Ferroptosis holds significant potential for application in cancer therapy. However, ferroptosis inducers are not cell-specific and can cause phospholipid peroxidation in both tumor and non-tumor cells. This limitation greatly restricts the use of ferroptosis therapy as a safe and effective anticancer strategy. Our previous study demonstrated that macrophages can engulf ferroptotic cells through Toll-like receptor 2 (TLR2). Despite this advancement, the precise mechanism by which phospholipid peroxidation in macrophages affects their phagocytotic capability during treatment of tumors with ferroptotic agents is still unknown. Here, we utilized flow sorting combined with redox phospholipidomics to determine that phospholipid peroxidation in tumor microenvironment (TME) macrophages impaired the macrophages ability to eliminate ferroptotic tumor cells by phagocytosis, ultimately fostering tumor resistance to ferroptosis therapy. Mechanistically, the accumulation of phospholipid peroxidation in the macrophage endoplasmic reticulum (ER) repressed TLR2 trafficking to the plasma membrane and caused its retention in the ER by disrupting the interaction between TLR2 and its chaperone CNPY3. Subsequently, this ER-retained TLR2 recruited E3 ligase MARCH6 and initiated the proteasome-dependent degradation. Using redox phospholipidomics, we identified 1-steaoryl-2-15-HpETE-sn-glycero-3-phosphatidylethanolamine (SAPE-OOH) as the crucial mediator of these effects. Conclusively, our discovery elucidates a novel molecular mechanism underlying macrophage phospholipid peroxidation-induced tumor resistance to ferroptosis therapy and highlights the TLR2-MARCH6 axis as a potential therapeutic target for cancer therapy.

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Fig. 1: Phospholipid peroxidation of macrophages provokes tumor resistance to ferroptosis therapy.
Fig. 2: Phospholipid peroxidation of macrophages impairs phagocytic clearance of ferroptotic tumor cells.
Fig. 3: TLR2 plays an indispensable role in phospholipid peroxidation-provoked impairment of ferroptotic cell clearance and tumor resistance to ferroptosis therapy.
Fig. 4: Oxygenated PE disrupts the translocation of TLR2 from ER to PM by inhibiting the interaction between TLR2 and CNPY3.
Fig. 5: OxPEs in ER mediates proteasome-dependent degradation of TLR2.
Fig. 6: Identification of MARCH6 as a key E3 ligase in oxPEs-induced ubiquitin-dependent degradation of TLR2.
Fig. 7: SAPE-OOH serves as a crucial oxidized phospholipid for TLR2 ubiquitination.
Fig. 8: TLR2 agonist enhances the antitumor efficacy of ferroptosis inducer.

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Data are available upon reasonable request.

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Acknowledgements

We would like to thank Dr. Yong Jiang (Southern Medical University, Guangzhou, China) for Tlr2 KO C57BL/6 J mice (Jackson Laboratory, #004650), Dr. Kui Cheng (Southern Medical University) for providing SMU-Z1, Dr. Zhinan Yin (Jinan University, Guangzhou, China) for Lyz2cre C57BL/6 J mice (Jackson Laboratory, #004781).

Funding

This work was supported, in part, by Natural Science Foundation of China (82125038, T2341004, 82004231, 81903821, 81973718, 82274403, 82305063, 82374327), Guangdong Basic and Applied Basic Research Foundation (2021B1515120023, 2020A1515110388, 2021A1515011297, 2023B1515040016, 2023A1515110306, 2024A1515011423), the Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program (2017BT01Y036) and GDUPS (2019), Innovation Team Project of Guangdong Provincial Department of Education (2020KCXTD003), Guangdong-Hong Kong-Macao Universities Joint Laboratory for the Internationalization of Traditional Chinese Medicine (2023LSYS002), Guangzhou Key Laboratory of Traditional Chinese Medicine & Disease Susceptibility (2024A03J090), Guangzhou Basic and Applied Basic Research Foundation (2024A03J090, 2024A04J4020), Medical Science and Technology Research Foundation of Guangdong Province (A2023044), Fellowship of China Postdoctoral Science Foundation (2022TQ0122, 2023M731327, 2023M731326, 2024T170345), and Lift Project of Guangdong Second Provincial General Hospital (TJGC-2022002, 2022BSGZ007).

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  1. These authors contributed equally: Xiang Luo, Hai-Biao Gong.

Authors and Affiliations

  1. Guangdong Second Provincial General Hospital/Guangdong Engineering Research Center of Chinese Medicine & Disease Susceptibility/Guangzhou Key Laboratory of Traditional Chinese Medicine & Disease Susceptibility/Guangdong-Hong Kong-Macao Universities Joint Laboratory for the Internationalization of Traditional Chinese Medicine/International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry of Education (MOE)/Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research/State Key Laboratory of Bioactive Molecules and Druggability Assessment/Integrated Chinese and Western Medicine Postdoctoral Research Station, Jinan University, Guangzhou, 510632, China

    Xiang Luo, Hai-Biao Gong, Zi-Chun Li, Dong-Dong Li, Zi-Xuan Li, Jie Sun, Chang-Yu Yan, Shu-Rui Chen, Rong Wang, Wan-Yang Sun, Hiroshi Kurihara, Wen-Jun Duan, Lei Liang, Wen Jin, Yan-Ping Wu, Rong-Rong He & Yi-Fang Li

  2. State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, 999078, China

    Rui-Ting Huang & Rong-Rong He

  3. Department of Materials Science and Engineering, College of Chemistry and Materials Science, Jinan University, Guangzhou, 511443, China

    Yue Feng & Mingxian Liu

  4. Shanghai Institute for Biomedical and Pharmaceutical Technologies, National Health Commission Key Laboratory of Reproduction Regulation, Shanghai, China

    Yun-Feng Cao

  5. School of Chinese Materia Medica and Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan University of Chinese Medicine, Kunming, 650500, China

    Rong Wang, Feng Huang & Rong-Rong He

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Contributions

R-RH, Y-FL and Y-PW conceived and designed the experiments. XL performed the experiments and prepared the manuscript. Z-CL, D-DL, Z-XL, JS, C-YY, R-TH, S-RC assisted the experiments and data analysis. H-BG, RW, W-YS conducted and analyzed the LC–MS/MS-based experiments. YF and ML conducted the synthesis of mPEG-PCL-SSZ micelles. Y-FC supported the LC-MS platform and consulted the data analysis. LL, W-JD, FH, HK and WJ advised the project and revised the manuscript. Y-FL, R-RH and Y-PW revised and approved the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yan-Ping Wu, Rong-Rong He or Yi-Fang Li.

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All animal experiment protocols were undertaken in accordance with the National Institutes of Health’s Guide for the Care and Use of Laboratory mice and were approved by the Laboratory Animal Ethic Committee of Jinan University (approval number: IACUC-20220613-09).

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Luo, X., Gong, HB., Li, ZC. et al. Phospholipid peroxidation in macrophage confers tumor resistance by suppressing phagocytic capability towards ferroptotic cells. Cell Death Differ 31, 1184–1201 (2024). https://doi.org/10.1038/s41418-024-01351-0

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