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 Xiao; Oscar C. Salgado; Nicholas Jarjour; Kristin A. Hogquist; Xavier Revelo; Alma L. Burlingame; Xiang Gao; Jakob von Moltke; Zhaoyu Lin; Hai-Bin Ruan

doi:10.1016/j.immuni.2022.03.009

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 journal › Article › peer-review

26 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 language	English (US)
Pages (from-to)	623-638.e5
Journal	Immunity
Volume	55
Issue number	4
DOIs	https://doi.org/10.1016/j.immuni.2022.03.009
State	Published - 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

PubMed: MeSH publication types

Journal Article

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.immuni.2022.03.009

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Zhao, M., Ren, K., Xiong, X., Xin, Y., Zou, Y., Maynard, J. C., Kim, A., Battist, A. P., Koneripalli, N., Wang, Y., Chen, Q., Xin, R., Yang, C., Huang, R., Yu, J., Huang, Z., Zhang, Z., Wang, H., Wang, D., ... Ruan, H-B. (2022). Epithelial STAT6 O-GlcNAcylation drives a concerted anti-helminth alarmin response dependent on tuft cell hyperplasia and Gasdermin C. Immunity, 55(4), 623-638.e5. https://doi.org/10.1016/j.immuni.2022.03.009

Zhao, M, Ren, K, Xiong, X, Xin, Y, Zou, Y, Maynard, JC, Kim, A, Battist, AP, Koneripalli, N, Wang, Y, Chen, Q, Xin, R, Yang, C, Huang, R, Yu, J, Huang, Z, Zhang, Z, Wang, H, Wang, D, Xiao, Y, Salgado, OC, Jarjour, N , Hogquist, KA , Revelo, X, Burlingame, AL, Gao, X, von Moltke, J, Lin, Z & Ruan, H-B 2022, 'Epithelial STAT6 O-GlcNAcylation drives a concerted anti-helminth alarmin response dependent on tuft cell hyperplasia and Gasdermin C', Immunity, vol. 55, no. 4, pp. 623-638.e5. https://doi.org/10.1016/j.immuni.2022.03.009

@article{9e524b966a7343e8a1ff7aaff9ef8aa1,

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

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.",

keywords = "Gasdermin, IL-10, IL-25, IL-33, O-GlcNAc, OGT, STAT6, Tuft cell, colitis, goblet cell",

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

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: {\textcopyright} 2022 Elsevier Inc.",

year = "2022",

month = apr,

day = "12",

doi = "10.1016/j.immuni.2022.03.009",

language = "English (US)",

volume = "55",

pages = "623--638.e5",

journal = "Immunity",

issn = "1074-7613",

publisher = "Cell Press",

number = "4",

}

TY - JOUR

T1 - Epithelial STAT6 O-GlcNAcylation drives a concerted anti-helminth alarmin response dependent on tuft cell hyperplasia and Gasdermin C

AU - Zhao, Ming

AU - Ren, Kaiqun

AU - Xiong, Xiwen

AU - Xin, Yue

AU - Zou, Yujie

AU - Maynard, Jason C.

AU - Kim, Angela

AU - Battist, Alexander P.

AU - Koneripalli, Navya

AU - Wang, Yusu

AU - Chen, Qianyue

AU - Xin, Ruyue

AU - Yang, Chenyan

AU - Huang, Rong

AU - Yu, Jiahui

AU - Huang, Zan

AU - Zhang, Zengdi

AU - Wang, Haiguang

AU - Wang, Daoyuan

AU - Xiao, Yihui

AU - Salgado, Oscar C.

AU - Jarjour, Nicholas

AU - Hogquist, Kristin A.

AU - Revelo, Xavier

AU - Burlingame, Alma L.

AU - Gao, Xiang

AU - von Moltke, Jakob

AU - Lin, Zhaoyu

AU - Ruan, Hai-Bin

N1 - 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.

PY - 2022/4/12

Y1 - 2022/4/12

N2 - 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.

AB - 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.

KW - Gasdermin

KW - IL-10

KW - IL-25

KW - IL-33

KW - O-GlcNAc

KW - OGT

KW - STAT6

KW - Tuft cell

KW - colitis

KW - goblet cell

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UR - http://www.scopus.com/inward/citedby.url?scp=85127492845&partnerID=8YFLogxK

U2 - 10.1016/j.immuni.2022.03.009

DO - 10.1016/j.immuni.2022.03.009

M3 - Article

C2 - 35385697

AN - SCOPUS:85127492845

SN - 1074-7613

VL - 55

SP - 623-638.e5

JO - Immunity

JF - Immunity

IS - 4

ER -

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

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

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