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mTORC2 acts as a gatekeeper for mTORC1 deficiency-mediated impairments in ILC3 development

Acta Pharmacologica Sinica volume 44, pages 2243–2252 (2023)Cite this article

Abstract

Group 3 innate lymphoid cells (ILC3s) are mediators of intestinal immunity and barrier function. Recent studies have investigated the role of the mammalian target of rapamycin complex (mTOR) in ILC3s, whereas the mTORC1-related mechanisms and crosstalk between mTORC1 and mTORC2 involved in regulating ILC3 homeostasis remain unknown. In this study, we found that mTORC1 but not mTORC2 was critical in ILC3 development, IL-22 production, and ILC3-mediated intestinal homeostasis. Single-cell RNA sequencing revealed that mTORC1 deficiency led to disruption of ILC3 heterogeneity, showing an increase in differentiation into ILC1-like phenotypes. Mechanistically, mTORC1 deficiency decreased the expression of NFIL3, which is a critical transcription factor responsible for ILC3 development. The activities of both mTORC1 and mTORC2 were increased in wild-type ILC3s after activation by IL-23, whereas inhibition of mTORC1 by Raptor deletion or rapamycin treatment resulted in increased mTORC2 activity. Previous studies have demonstrated that S6K, the main downstream target of mTORC1, can directly phosphorylate Rictor to dampen mTORC2 activity. Our data found that inhibition of mTORC1 activity by rapamycin reduced Rictor phosphorylation in ILC3s. Reversing the increased mTORC2 activity via heterozygous or homozygous knockout of Rictor in Raptor-deleted ILC3s resulted in severe ILC3 loss and complete susceptibility to intestinal infection in mice with mTORC1 deficiency (100% mortality). Thus, mTORC1 acts as a rheostat of ILC3 heterogeneity, and mTORC2 protects ILC3s from severe loss of cells and immune activity against intestinal infection when mTORC1 activity is diminished.

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Fig. 1: mTORC1 but not mTORC2 is essential for ILC3 development, IL-22 production and ILC3-mediated host defense.
Fig. 2: Single-cell RNA sequencing (scRNA-seq) reveals mTORC1 as a regulator of ILC3 heterogeneity.
Fig. 3: mTORC1 deficiency results in reduced Nfil3 expression in ILC3s.
Fig. 4: Genetic deficiency or chemical inhibition of mTORC1 activity results in increased mTORC2 activity via reduced Rictor phosphorylation.
Fig. 5: Upregulated mTORC2 activity protects ILC3s from the severe reduction in cell numbers and loss of the intestinal defense caused by mTORC1 deficiency.

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Data availability

All data are included in the manuscript. The datasets analyzed in the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This study was supported by funding from the National Natural Science Foundation of China (Nos. 81874313 and 81922068 to YCD; No. 81703521 to YFD; No. 81900055 to HYP; and No. 82070571 to YLW). This study was also supported by funding from the Natural Science Foundation of Hunan Province (No. 2023JJ30320 to YFD and No. 2021JJ40274 to HYP) and the Natural Science Foundation of Changsha, Hunan Province (No. kq2208090 to YFD) and the Project of Army Medical University (2109XQ04 to YCD). This work was also supported by funding from the Science Foundation of Hunan Children’s Hospital.

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  1. These authors contributed equally: Ya-fei Deng, Shu-ting Wu

Authors and Affiliations

  1. Pediatrics Research Institute of Hunan Province and Hunan Provincial Key Laboratory of Children’s Emergency Medicine, Hunan Children’s Hospital, Changsha, 410007, China

    Ya-fei Deng, Shu-ting Wu, Hong-yan Peng, Ya-na Li, Lan-lan Huang, Pei-wen Xiong, Song-yang Li, Qing-lan Yang, Li-li Wang, Li-ping Li, Xiu-lan Lu & Zheng-hui Xiao

  2. Department of Clinical Hematology, College of Pharmacy and Laboratory Medicine Science, Army Medical University, Chongqing, 400038, China

    Ya-fei Deng, Yao Yang, Meng Meng, Xiao-hui Li & You-cai Deng

  3. Department of Hematology and Hematopoietic Cell Transplantation, City of Hope National Medical Center, Los Angeles, CA, USA

    Lei Tian & Jian-hua Yu

  4. Department of Clinical Pharmacy, Weifang Traditional Chinese Hospital, Weifang, 261041, China

    Xiao-yao Li

  5. Department of Gastroenterology, Chongqing Key Laboratory of Digestive Malignancies, Daping Hospital, Army Medical University, Chongqing, 400042, China

    Yan-ling Wei

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Contributions

The work presented was performed in collaboration by all authors. YFD designed and performed the experiments, analyzed the data, and wrote the manuscript. STW, HYP, YNL, YY, MM, LLH, PWX, SYL, QLY, LLW and XYL performed the experiments and analyzed the data. LPL, XLL and XHL designed the research and supervised the study. YLW, ZHX, JHY and YCD revised the concept, designed the research, supervised the study, and wrote the manuscript.

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Correspondence to Yan-ling Wei, Zheng-hui Xiao, Jian-hua Yu or You-cai Deng.

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Deng, Yf., Wu, St., Peng, Hy. et al. mTORC2 acts as a gatekeeper for mTORC1 deficiency-mediated impairments in ILC3 development. Acta Pharmacol Sin 44, 2243–2252 (2023). https://doi.org/10.1038/s41401-023-01120-8

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