Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut

Jinzhi Duan; Juan D. Matute; Lukas W. Unger; Thomas Hanley; Alexandra Schnell; Xi Lin; Niklas Krupka; Paul Griebel; Conner Lambden; Brandon Sit; Joep Grootjans; Michal Pyzik; Felix Sommer; Sina Kaiser; Maren Falk-Paulsen; Helmut Grasberger; John Y. Kao; Tobias Fuhrer; Hai Li; Donggi Paik; Yunjin Lee; Samuel Refetoff; Jonathan N. Glickman; Adrienne W. Paton; Lynn Bry; James C. Paton; Uwe Sauer; Andrew J. Macpherson; Philip Rosenstiel; Vijay K. Kuchroo; Matthew K. Waldor; Jun R. Huh; Arthur Kaser; Richard S. Blumberg

doi:https://doi.org/10.1016/j.immuni.2023.02.018

Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut

Jinzhi Duan, Juan D. Matute, Lukas W. Unger, Thomas Hanley, Alexandra Schnell, Xi Lin, Niklas Krupka, Paul Griebel, Conner Lambden, Brandon Sit, Joep Grootjans, Michal Pyzik, Felix Sommer, Sina Kaiser, Maren Falk-Paulsen, Helmut Grasberger, John Y. Kao, Tobias Fuhrer, Hai Li, Donggi PaikYunjin Lee, Samuel Refetoff, Jonathan N. Glickman, Adrienne W. Paton, Lynn Bry, James C. Paton, Uwe Sauer, Andrew J. Macpherson, Philip Rosenstiel, Vijay K. Kuchroo, Matthew K. Waldor, Jun R. Huh, Arthur Kaser, Richard S. Blumberg

Research output: Contribution to journal › Article › Academic › peer-review

11 Citations (Scopus)

Abstract

Intestinal IL-17-producing T helper (Th17) cells are dependent on adherent microbes in the gut for their development. However, how microbial adherence to intestinal epithelial cells (IECs) promotes Th17 cell differentiation remains enigmatic. Here, we found that Th17 cell-inducing gut bacteria generated an unfolded protein response (UPR) in IECs. Furthermore, subtilase cytotoxin expression or genetic removal of X-box binding protein 1 (Xbp1) in IECs caused a UPR and increased Th17 cells, even in antibiotic-treated or germ-free conditions. Mechanistically, UPR activation in IECs enhanced their production of both reactive oxygen species (ROS) and purine metabolites. Treating mice with N-acetyl-cysteine or allopurinol to reduce ROS production and xanthine, respectively, decreased Th17 cells that were associated with an elevated UPR. Th17-related genes also correlated with ER stress and the UPR in humans with inflammatory bowel disease. Overall, we identify a mechanism of intestinal Th17 cell differentiation that emerges from an IEC-associated UPR.

Original language	English
Pages (from-to)	1115-1131.e9
Journal	Immunity
Volume	56
Issue number	5
Early online date	2023
DOIs	https://doi.org/10.1016/j.immuni.2023.02.018
Publication status	Published - 9 May 2023

Keywords

Citrobacter rodentium
ROS signals
TH17 cells
commensal bacterial
epithelial endoplasmic reticulum stress
inflammatory bowel disease
purine metabolism

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https://doi.org/10.1016/j.immuni.2023.02.018

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Duan, J., Matute, J. D., Unger, L. W., Hanley, T., Schnell, A., Lin, X., Krupka, N., Griebel, P., Lambden, C., Sit, B., Grootjans, J., Pyzik, M., Sommer, F., Kaiser, S., Falk-Paulsen, M., Grasberger, H., Kao, J. Y., Fuhrer, T., Li, H., ... Blumberg, R. S. (2023). Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut. Immunity, 56(5), 1115-1131.e9. https://doi.org/10.1016/j.immuni.2023.02.018

@article{d1ef37b3db6c4e3484559d2cea69ee42,

title = "Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut",

abstract = "Intestinal IL-17-producing T helper (Th17) cells are dependent on adherent microbes in the gut for their development. However, how microbial adherence to intestinal epithelial cells (IECs) promotes Th17 cell differentiation remains enigmatic. Here, we found that Th17 cell-inducing gut bacteria generated an unfolded protein response (UPR) in IECs. Furthermore, subtilase cytotoxin expression or genetic removal of X-box binding protein 1 (Xbp1) in IECs caused a UPR and increased Th17 cells, even in antibiotic-treated or germ-free conditions. Mechanistically, UPR activation in IECs enhanced their production of both reactive oxygen species (ROS) and purine metabolites. Treating mice with N-acetyl-cysteine or allopurinol to reduce ROS production and xanthine, respectively, decreased Th17 cells that were associated with an elevated UPR. Th17-related genes also correlated with ER stress and the UPR in humans with inflammatory bowel disease. Overall, we identify a mechanism of intestinal Th17 cell differentiation that emerges from an IEC-associated UPR.",

keywords = "Citrobacter rodentium, ROS signals, TH17 cells, commensal bacterial, epithelial endoplasmic reticulum stress, inflammatory bowel disease, purine metabolism",

author = "Jinzhi Duan and Matute, {Juan D.} and Unger, {Lukas W.} and Thomas Hanley and Alexandra Schnell and Xi Lin and Niklas Krupka and Paul Griebel and Conner Lambden and Brandon Sit and Joep Grootjans and Michal Pyzik and Felix Sommer and Sina Kaiser and Maren Falk-Paulsen and Helmut Grasberger and Kao, {John Y.} and Tobias Fuhrer and Hai Li and Donggi Paik and Yunjin Lee and Samuel Refetoff and Glickman, {Jonathan N.} and Paton, {Adrienne W.} and Lynn Bry and Paton, {James C.} and Uwe Sauer and Macpherson, {Andrew J.} and Philip Rosenstiel and Kuchroo, {Vijay K.} and Waldor, {Matthew K.} and Huh, {Jun R.} and Arthur Kaser and Blumberg, {Richard S.}",

note = "Funding Information: We thank L.H. Glimcher for providing C57BL/6J Xbp1fl/fl mice. We thank the members of the Blumberg lab, the Massachusetts host-microbiome center, Brigham and Women's Hospital Single Cell Genomics Core, the Harvard genome modification facility, and Brigham and Women's Hospital animal facility for their support of this project. We thank M.F. Paulsen, J.A. West, and K. Ramshorn for their kind help on this project. We thank all the patients who joined in the PROTECT and RISK studies. We acknowledge support by the NIHR Cambridge BRC. This work was supported by NIH grants DK044319, DK051362, DK053056, and DK088199; the NIH Harvard Digestive Diseases Center (HDDC) DK034854 (R.S.B. and J.D.M.); CCF Research Fellowship Award #707702 and the NIH Pediatric Scientist Development Program K12HD000850 (J.D.M.); NIH grant DK117565 (J.Y.K. and H.G.); NIH grant DK110559 (J.R.H.); NIH grant DK015070 (S.R.); Austrian Science Fund FWF J 4396 (L.W.U.); the Wellcome Trust (senior investigator award 106260/Z/14/Z and 222497/Z/21/Z); the European Research Council (HORIZON2020/ERC grant agreement no. 648889) (A.K.); the DFG individual grant SO1141/10-1; the DFG Research Unit FOR5042 “miTarget - The Microbiome as a Target in Inflammatory Bowel Diseases” (project P5) (F.S.); the DFG Cluster of Excellence 2167 Precision Medicine in Chronic Inflammation; the BMBF project iTREAT (SP5); and the EU H2020 grant SYSCID (contract no. 733100) (P.R.). J.D. J.D.M. J.R.H. A.K. and R.S.B. conceived, designed, and interpreted data. J.D. J.D.M. T.H. N.K. P.G. D.P. Y.L. L.B. F.S. S.K. M.F.-P. J.N.G. and M.P. performed most mouse and in vitro experiments. J.D. J.G. H.L. T.F. U.S. A.J.M. L.W.U. and A.K. performed all the metabolite analyses. B.S. and M.K.W. generated the C. rodentium Δeae strain. X.L. A.S. C.L. and V.K.K. performed the Nanostring nCounter sequence and single-cell RNA sequence analysis. J.C.P. and A.W.P. provided the SubA construct and antibody. P.R. and F.S. generated Duox2fl/fl mice. H.G. J.Y.K. and S.R. provided Duoxa−/− mice. J.D. J.D.M. A.S. C.L. L.W.U. M.P. A.K. J.R.H. and R.S.B. wrote the manuscript with the help of all other authors. The authors declare no competing interests. Funding Information: We thank L.H. Glimcher for providing C57BL/6J Xbp1 fl/fl mice. We thank the members of the Blumberg lab, the Massachusetts host-microbiome center, Brigham and Women{\textquoteright}s Hospital Single Cell Genomics Core, the Harvard genome modification facility, and Brigham and Women{\textquoteright}s Hospital animal facility for their support of this project. We thank M.F. Paulsen, J.A. West, and K. Ramshorn for their kind help on this project. We thank all the patients who joined in the PROTECT and RISK studies. We acknowledge support by the NIHR Cambridge BRC . This work was supported by NIH grants DK044319 , DK051362 , DK053056 , and DK088199 ; the NIH Harvard Digestive Diseases Center (HDDC) DK034854 (R.S.B. and J.D.M.); CCF Research Fellowship Award #707702 and the NIH Pediatric Scientist Development Program K12HD000850 (J.D.M.); NIH grant DK117565 (J.Y.K. and H.G.); NIH grant DK110559 (J.R.H.); NIH grant DK015070 (S.R.); Austrian Science Fund FWF J 4396 (L.W.U.); the Wellcome Trust (senior investigator award 106260/Z/14/Z and 222497/Z/21/Z); the European Research Council (HORIZON2020/ ERC grant agreement no. 648889) (A.K.); the DFG individual grant SO1141/10-1; the DFG Research Unit FOR5042 “miTarget - The Microbiome as a Target in Inflammatory Bowel Diseases” (project P5) (F.S.); the DFG Cluster of Excellence 2167 Precision Medicine in Chronic Inflammation; the BMBF project iTREAT (SP5); and the EU H2020 grant SYSCID (contract no. 733100) (P.R.). Publisher Copyright: {\textcopyright} 2023 Elsevier Inc.",

year = "2023",

month = may,

day = "9",

doi = "https://doi.org/10.1016/j.immuni.2023.02.018",

language = "English",

volume = "56",

pages = "1115--1131.e9",

journal = "Immunity",

issn = "1074-7613",

publisher = "Cell Press",

number = "5",

}

Duan, J, Matute, JD, Unger, LW, Hanley, T, Schnell, A, Lin, X, Krupka, N, Griebel, P, Lambden, C, Sit, B, Grootjans, J, Pyzik, M, Sommer, F, Kaiser, S, Falk-Paulsen, M, Grasberger, H, Kao, JY, Fuhrer, T, Li, H, Paik, D, Lee, Y, Refetoff, S, Glickman, JN, Paton, AW, Bry, L, Paton, JC, Sauer, U, Macpherson, AJ, Rosenstiel, P, Kuchroo, VK, Waldor, MK, Huh, JR, Kaser, A & Blumberg, RS 2023, 'Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut', Immunity, vol. 56, no. 5, pp. 1115-1131.e9. https://doi.org/10.1016/j.immuni.2023.02.018

TY - JOUR

T1 - Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut

AU - Duan, Jinzhi

AU - Matute, Juan D.

AU - Unger, Lukas W.

AU - Hanley, Thomas

AU - Schnell, Alexandra

AU - Lin, Xi

AU - Krupka, Niklas

AU - Griebel, Paul

AU - Lambden, Conner

AU - Sit, Brandon

AU - Grootjans, Joep

AU - Pyzik, Michal

AU - Sommer, Felix

AU - Kaiser, Sina

AU - Falk-Paulsen, Maren

AU - Grasberger, Helmut

AU - Kao, John Y.

AU - Fuhrer, Tobias

AU - Li, Hai

AU - Paik, Donggi

AU - Lee, Yunjin

AU - Refetoff, Samuel

AU - Glickman, Jonathan N.

AU - Paton, Adrienne W.

AU - Bry, Lynn

AU - Paton, James C.

AU - Sauer, Uwe

AU - Macpherson, Andrew J.

AU - Rosenstiel, Philip

AU - Kuchroo, Vijay K.

AU - Waldor, Matthew K.

AU - Huh, Jun R.

AU - Kaser, Arthur

AU - Blumberg, Richard S.

N1 - Funding Information: We thank L.H. Glimcher for providing C57BL/6J Xbp1fl/fl mice. We thank the members of the Blumberg lab, the Massachusetts host-microbiome center, Brigham and Women's Hospital Single Cell Genomics Core, the Harvard genome modification facility, and Brigham and Women's Hospital animal facility for their support of this project. We thank M.F. Paulsen, J.A. West, and K. Ramshorn for their kind help on this project. We thank all the patients who joined in the PROTECT and RISK studies. We acknowledge support by the NIHR Cambridge BRC. This work was supported by NIH grants DK044319, DK051362, DK053056, and DK088199; the NIH Harvard Digestive Diseases Center (HDDC) DK034854 (R.S.B. and J.D.M.); CCF Research Fellowship Award #707702 and the NIH Pediatric Scientist Development Program K12HD000850 (J.D.M.); NIH grant DK117565 (J.Y.K. and H.G.); NIH grant DK110559 (J.R.H.); NIH grant DK015070 (S.R.); Austrian Science Fund FWF J 4396 (L.W.U.); the Wellcome Trust (senior investigator award 106260/Z/14/Z and 222497/Z/21/Z); the European Research Council (HORIZON2020/ERC grant agreement no. 648889) (A.K.); the DFG individual grant SO1141/10-1; the DFG Research Unit FOR5042 “miTarget - The Microbiome as a Target in Inflammatory Bowel Diseases” (project P5) (F.S.); the DFG Cluster of Excellence 2167 Precision Medicine in Chronic Inflammation; the BMBF project iTREAT (SP5); and the EU H2020 grant SYSCID (contract no. 733100) (P.R.). J.D. J.D.M. J.R.H. A.K. and R.S.B. conceived, designed, and interpreted data. J.D. J.D.M. T.H. N.K. P.G. D.P. Y.L. L.B. F.S. S.K. M.F.-P. J.N.G. and M.P. performed most mouse and in vitro experiments. J.D. J.G. H.L. T.F. U.S. A.J.M. L.W.U. and A.K. performed all the metabolite analyses. B.S. and M.K.W. generated the C. rodentium Δeae strain. X.L. A.S. C.L. and V.K.K. performed the Nanostring nCounter sequence and single-cell RNA sequence analysis. J.C.P. and A.W.P. provided the SubA construct and antibody. P.R. and F.S. generated Duox2fl/fl mice. H.G. J.Y.K. and S.R. provided Duoxa−/− mice. J.D. J.D.M. A.S. C.L. L.W.U. M.P. A.K. J.R.H. and R.S.B. wrote the manuscript with the help of all other authors. The authors declare no competing interests. Funding Information: We thank L.H. Glimcher for providing C57BL/6J Xbp1 fl/fl mice. We thank the members of the Blumberg lab, the Massachusetts host-microbiome center, Brigham and Women’s Hospital Single Cell Genomics Core, the Harvard genome modification facility, and Brigham and Women’s Hospital animal facility for their support of this project. We thank M.F. Paulsen, J.A. West, and K. Ramshorn for their kind help on this project. We thank all the patients who joined in the PROTECT and RISK studies. We acknowledge support by the NIHR Cambridge BRC . This work was supported by NIH grants DK044319 , DK051362 , DK053056 , and DK088199 ; the NIH Harvard Digestive Diseases Center (HDDC) DK034854 (R.S.B. and J.D.M.); CCF Research Fellowship Award #707702 and the NIH Pediatric Scientist Development Program K12HD000850 (J.D.M.); NIH grant DK117565 (J.Y.K. and H.G.); NIH grant DK110559 (J.R.H.); NIH grant DK015070 (S.R.); Austrian Science Fund FWF J 4396 (L.W.U.); the Wellcome Trust (senior investigator award 106260/Z/14/Z and 222497/Z/21/Z); the European Research Council (HORIZON2020/ ERC grant agreement no. 648889) (A.K.); the DFG individual grant SO1141/10-1; the DFG Research Unit FOR5042 “miTarget - The Microbiome as a Target in Inflammatory Bowel Diseases” (project P5) (F.S.); the DFG Cluster of Excellence 2167 Precision Medicine in Chronic Inflammation; the BMBF project iTREAT (SP5); and the EU H2020 grant SYSCID (contract no. 733100) (P.R.). Publisher Copyright: © 2023 Elsevier Inc.

PY - 2023/5/9

Y1 - 2023/5/9

N2 - Intestinal IL-17-producing T helper (Th17) cells are dependent on adherent microbes in the gut for their development. However, how microbial adherence to intestinal epithelial cells (IECs) promotes Th17 cell differentiation remains enigmatic. Here, we found that Th17 cell-inducing gut bacteria generated an unfolded protein response (UPR) in IECs. Furthermore, subtilase cytotoxin expression or genetic removal of X-box binding protein 1 (Xbp1) in IECs caused a UPR and increased Th17 cells, even in antibiotic-treated or germ-free conditions. Mechanistically, UPR activation in IECs enhanced their production of both reactive oxygen species (ROS) and purine metabolites. Treating mice with N-acetyl-cysteine or allopurinol to reduce ROS production and xanthine, respectively, decreased Th17 cells that were associated with an elevated UPR. Th17-related genes also correlated with ER stress and the UPR in humans with inflammatory bowel disease. Overall, we identify a mechanism of intestinal Th17 cell differentiation that emerges from an IEC-associated UPR.

AB - Intestinal IL-17-producing T helper (Th17) cells are dependent on adherent microbes in the gut for their development. However, how microbial adherence to intestinal epithelial cells (IECs) promotes Th17 cell differentiation remains enigmatic. Here, we found that Th17 cell-inducing gut bacteria generated an unfolded protein response (UPR) in IECs. Furthermore, subtilase cytotoxin expression or genetic removal of X-box binding protein 1 (Xbp1) in IECs caused a UPR and increased Th17 cells, even in antibiotic-treated or germ-free conditions. Mechanistically, UPR activation in IECs enhanced their production of both reactive oxygen species (ROS) and purine metabolites. Treating mice with N-acetyl-cysteine or allopurinol to reduce ROS production and xanthine, respectively, decreased Th17 cells that were associated with an elevated UPR. Th17-related genes also correlated with ER stress and the UPR in humans with inflammatory bowel disease. Overall, we identify a mechanism of intestinal Th17 cell differentiation that emerges from an IEC-associated UPR.

KW - Citrobacter rodentium

KW - ROS signals

KW - TH17 cells

KW - commensal bacterial

KW - epithelial endoplasmic reticulum stress

KW - inflammatory bowel disease

KW - purine metabolism

UR - http://www.scopus.com/inward/record.url?scp=85151571929&partnerID=8YFLogxK

U2 - https://doi.org/10.1016/j.immuni.2023.02.018

DO - https://doi.org/10.1016/j.immuni.2023.02.018

M3 - Article

C2 - 36917985

SN - 1074-7613

VL - 56

SP - 1115-1131.e9

JO - Immunity

JF - Immunity

IS - 5

ER -

Endoplasmic reticulum stress in the intestinal epithelium initiates purine metabolite synthesis and promotes Th17 cell differentiation in the gut

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