Survival signal REG3α prevents crypt apoptosis to control acute gastrointestinal graft-versus-host disease
Dongchang Zhao, … , Pavan Reddy, James L.M. Ferrara
Dongchang Zhao, … , Pavan Reddy, James L.M. Ferrara
Published August 14, 2018
Citation Information: J Clin Invest. 2018;128(11):4970-4979. https://doi.org/10.1172/JCI99261.
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Concise Communication Immunology

Survival signal REG3α prevents crypt apoptosis to control acute gastrointestinal graft-versus-host disease

Abstract

Graft-versus-host disease (GVHD) in the gastrointestinal (GI) tract remains the major cause of morbidity and nonrelapse mortality after BM transplantation (BMT). The Paneth cell protein regenerating islet-derived 3α (REG3α) is a biomarker specific for GI GVHD. REG3α serum levels rose in the systematic circulation as GVHD progressively destroyed Paneth cells and reduced GI epithelial barrier function. Paradoxically, GVHD suppressed intestinal REG3γ (the mouse homolog of human REG3α), and the absence of REG3γ in BMT recipients intensified GVHD but did not change the composition of the microbiome. IL-22 administration restored REG3γ production and prevented apoptosis of both intestinal stem cells (ISCs) and Paneth cells, but this protection was completely abrogated in Reg3g−/− mice. In vitro, addition of REG3α reduced the apoptosis of colonic cell lines. Strategies that increase intestinal REG3α/γ to promote crypt regeneration may offer a novel, nonimmunosuppressive approach for GVHD and perhaps for other diseases involving the ISC niche, such as inflammatory bowel disease.

Authors

Dongchang Zhao, Yeung-Hyen Kim, Seihwan Jeong, Joel K. Greenson, Mohammed S. Chaudhry, Matthias Hoepting, Erik R. Anderson, Marcel R.M. van den Brink, Jonathan U. Peled, Antonio L.C. Gomes, Ann E. Slingerland, Michael J. Donovan, Andrew C. Harris, John E. Levine, Umut Ozbek, Lora V. Hooper, Thaddeus S. Stappenbeck, Aaron Ver Heul, Ta-Chiang Liu, Pavan Reddy, James L.M. Ferrara

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Figure 8

REG3γ protects ISCs from apoptosis.

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REG3γ protects ISCs from apoptosis. (A–D) WT B6 and B6-Reg3g–/– (KO) mic...
(A–D) WT B6 and B6-Reg3g–/– (KO) mice (Lgr5-EGFP+) received PBS or IL-22 injections from day +1 after BMT from B6-Ly5.1 donors (GVHD –) or C3H.SW donors (GVHD +). On day +7 after BMT, small intestine crypt cells were analyzed by flow cytometry, and Reg3g mRNA was measured by qPCR. (A) Quantification of ISCs from mice in Figure 7. (B) Quantification of ISCs undergoing early apoptosis by annexin V+ staining in the mice from Figure 7. Data are combined from 3 separate experiments. (C) Quantification of EdU+ ISCs in syngeneic recipients (GVHD –: WT-PBS, empty black circles, n = 7; WT–IL-22, empty blue circles, n = 4; KO-PBS, empty red squares, n = 9; KO–IL-22, empty blue squares, n = 5), allogeneic recipients (GVHD +: WT-PBS, filled black circles, n = 8; WT–IL-22, filled blue circles, n = 9; KO-PBS, filled red squares, n = 13; KO–IL-22, filled blue squares, n = 11), and naive WT (downward triangles, n = 5) and KO (upward triangles, n = 5) controls. (D) Correlation of ISCs and Reg3g mRNA expression in WT mice from 2 of the 3 experiments shown in A (n = 25), as calculated by the Pearson correlation coefficient. (E–G) HT-29 cells were cultured with recombinant REG3α protein and apoptotic stimuli as described in Methods. Data are representative of 3 independent experiments. (E and F) Cell lysates were collected after 16-hour cell culture for Western blot analysis. Quantification of cleaved caspase-3 (E) and cleaved caspase-8 (F) expression. (G) Quantification of cell viability by CellTiter-Glo 2.0 assay after 24-hour cell culture. *P < 0.05, **P < 0.01, unpaired 2-tailed t test. Data are expressed as mean ± SEM. ND, not detectable.