Epithelial STAT6 O-GlcNAcylation drives a concerted anti-helminth alarmin response dependent on tuft cell hyperplasia and Gasdermin C

Ming Zhao, Kaiqun Ren, Xiwen Xiong, Yue Xin, Yujie Zou, Jason C. Maynard, Angela Kim, Alexander P. Battist, Navya Koneripalli, Yusu Wang, Qianyue Chen, Ruyue Xin, Chenyan Yang, Rong Huang, Jiahui Yu, Zan Huang, Zengdi Zhang, Haiguang Wang, Daoyuan Wang, Yihui XiaoOscar C. Salgado, Nicholas Jarjour, Kristin A. Hogquist, Xavier Revelo, Alma L. Burlingame, Xiang Gao, Jakob von Moltke, Zhaoyu Lin, Hai-Bin Ruan

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

The epithelium is an integral component of mucosal barrier and host immunity. Following helminth infection, the intestinal epithelial cells secrete “alarmin” cytokines, such as interleukin-25 (IL-25) and IL-33, to initiate the type 2 immune responses for helminth expulsion and tolerance. However, it is unknown how helminth infection and the resulting cytokine milieu drive epithelial remodeling and orchestrate alarmin secretion. Here, we report that epithelial O-linked N-Acetylglucosamine (O-GlcNAc) protein modification was induced upon helminth infections. By modifying and activating the transcription factor STAT6, O-GlcNAc transferase promoted the transcription of lineage-defining Pou2f3 in tuft cell differentiation and IL-25 production. Meanwhile, STAT6 O-GlcNAcylation activated the expression of Gsdmc family genes. The membrane pore formed by GSDMC facilitated the unconventional secretion of IL-33. GSDMC-mediated IL-33 secretion was indispensable for effective anti-helminth immunity and contributed to induced intestinal inflammation. Protein O-GlcNAcylation can be harnessed for future treatment of type 2 inflammation-associated human diseases.

Original languageEnglish (US)
Pages (from-to)623-638.e5
JournalImmunity
Volume55
Issue number4
DOIs
StatePublished - Apr 12 2022

Bibliographical note

Funding Information:
We thank Dr. Timothy C. Wang, Dr. Xiaoyong Yang, and Dr. Fred D. Finkelman for providing Dclk1-CreER mice, Rosa26-LSL-rOGT/mOGA mice, and Il4ra −/− mice, respectively. We thank Dr. Reinhard Hinterleitner of University of Pittsburg for providing DNA extracted from cecum of Tritrichomonas colonized mice. This work was supported by National Institute of Health ( R21 AI140109 , R56 AI162791 , and R01 AI139420 ) to H.-B.R.; National Natural Science Foundation of China ( U1904132 ) and Program for Science & Technology Innovation Talents in Higher Education of Henan Province, China ( 20HASTIT046 ) to X.X.; Natural Science Foundation of Hunan Province, China ( 2015JJ6072 and 2020JJ4441 ) and Research Foundation of Education Bureau of Hunan Province, China ( 20A304 ) to K.R.; Ministry of Science and Technology of China ( 2018YFA0801100 ) to L.Z.; Ministry of Science and Technology of China ( 2021YFF0702100 ) and National Natural Science Foundation of China ( 31971056 ) to X.G.; NIH R01 DK122056 to X.S.R.; NIH 1DP2AI136596 to J.v.M.; and the Miriam and Sheldon G. Adelson Medical Research Foundation grant to A.L.B. H.W and N.N.J. received American Heart Association Postdoctoral Fellowship and Damon Runyon Cancer Research Foundation Fellowship ( DRG-2427-21 ), respectively.

Funding Information:
We thank Dr. Timothy C. Wang, Dr. Xiaoyong Yang, and Dr. Fred D. Finkelman for providing Dclk1-CreER mice, Rosa26-LSL-rOGT/mOGA mice, and Il4ra?/? mice, respectively. We thank Dr. Reinhard Hinterleitner of University of Pittsburg for providing DNA extracted from cecum of Tritrichomonas colonized mice. This work was supported by National Institute of Health (R21 AI140109, R56 AI162791, and R01 AI139420) to H.-B.R.; National Natural Science Foundation of China (U1904132) and Program for Science & Technology Innovation Talents in Higher Education of Henan Province, China (20HASTIT046) to X.X.; Natural Science Foundation of Hunan Province, China (2015JJ6072 and 2020JJ4441) and Research Foundation of Education Bureau of Hunan Province, China (20A304) to K.R.; Ministry of Science and Technology of China (2018YFA0801100) to L.Z.; Ministry of Science and Technology of China (2021YFF0702100) and National Natural Science Foundation of China (31971056) to X.G.; NIH R01 DK122056 to X.S.R.; NIH 1DP2AI136596 to J.v.M.; and the Miriam and Sheldon G. Adelson Medical Research Foundation grant to A.L.B. H.W and N.N.J. received American Heart Association Postdoctoral Fellowship and Damon Runyon Cancer Research Foundation Fellowship (DRG-2427-21), respectively. M.Z. designed, performed, and analyzed most experiments, particularly STAT6- and GSDMC-related work. K.R. initiated the project and analyzed IEC?Ogt and Tuft?Ogt mice. X.X. Y. Xin, C.Y. R.H. and J.Y. generated and analyzed IECCA-STAT6-Tg and ISC?Ogt mice. Y.Z. Y.W. Q.C. R.X. X.G. and Z.L. generated Gsdmc-floxed mice and performed Gsdmc RNA ISH and Gsdmc?/?;Il10?/? colitis. J.C.M. and A.L.B. performed mass spectrometric identification of STAT6 O-GlcNAc sites. A.K. A.P.B. N.K. D.W. and Y. Xiao assisted genotyping, DSS colitis, and qPCR. Z.H. and Z.Z. constructed lentiviruses. H.W. O.C.S, K.A.H, and X.S.R. performed flow cytometry. N.N.J. performed Il4ra?/? infection. J.v.M. provided reagents and guidance to helminth infection and organoid culture. H.-B.R. conceived, designed, and performed experiments. M.Z. and H.-B.R. wrote the manuscript. The authors disclose no conflicts. We worked to ensure sex balance in the selection of non-human subjects. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in science.

Publisher Copyright:
© 2022 Elsevier Inc.

Keywords

  • Gasdermin
  • IL-10
  • IL-25
  • IL-33
  • O-GlcNAc
  • OGT
  • STAT6
  • Tuft cell
  • colitis
  • goblet cell

Fingerprint

Dive into the research topics of 'Epithelial STAT6 O-GlcNAcylation drives a concerted anti-helminth alarmin response dependent on tuft cell hyperplasia and Gasdermin C'. Together they form a unique fingerprint.

Cite this