R-spondin/YAP axis promotes gastric oxyntic gland regeneration and Helicobacter pylori–associated metaplasia in mice
Anne-Sophie Fischer, … , Frank Tacke, Michael Sigal
Anne-Sophie Fischer, … , Frank Tacke, Michael Sigal
Published September 13, 2022
Citation Information: J Clin Invest. 2022;132(21):e151363. https://doi.org/10.1172/JCI151363.
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Research Article Gastroenterology

R-spondin/YAP axis promotes gastric oxyntic gland regeneration and Helicobacter pylori–associated metaplasia in mice

Abstract

The stomach corpus epithelium is organized into anatomical units that consist of glands and pits. Mechanisms that control the cellular organization of corpus glands and enable their recovery upon injury are not well understood. R-spondin 3 (RSPO3) is a WNT-signaling enhancer that regulates stem cell behavior in different organs. Here, we investigated the function of RSPO3 in the corpus during homeostasis, upon chief and/or parietal cell loss, and during chronic Helicobacter pylori infection. Using organoid culture and conditional mouse models, we demonstrate that RSPO3 is a critical driver of secretory cell differentiation in the corpus gland toward parietal and chief cells, while its absence promoted pit cell differentiation. Acute loss of chief and parietal cells induced by high dose tamoxifen — or merely the depletion of LGR5+ chief cells — caused an upregulation of RSPO3 expression, which was required for the initiation of a coordinated regenerative response via the activation of yes-associated protein (YAP) signaling. This response enabled a rapid recovery of the injured secretory gland cells. However, in the context of chronic H. pylori infection, the R-spondin–driven regeneration was maintained long term, promoting severe glandular hyperproliferation and the development of premalignant metaplasia.

Authors

Anne-Sophie Fischer, Stefanie Müllerke, Alexander Arnold, Julian Heuberger, Hilmar Berger, Manqiang Lin, Hans-Joachim Mollenkopf, Jonas Wizenty, David Horst, Frank Tacke, Michael Sigal

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Figure 3

RSPO3 is upregulated upon HDT-driven gland injury and upon depletion of LGR5+ chief cells and promotes glandular regeneration.

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RSPO3 is upregulated upon HDT-driven gland injury and upon depletion of ...
(A, B, D, E) Representative images of sections from (A and B) Rspo3-WT and (D and E) Rspo3-KO mice treated with HDT and sacrificed 1, 3, or 7 days later and nontreated controls stained for (A and D) H/K-ATPase (white) and (B and E) GIF (green), GSII (red), and Ki67 (yellow). (C) qPCR for Rspo3 expression in Rspo3-WT mice treated with HDT and sacrificed on day 1 (n = 6), 3 (n = 4), or 7 (n = 4) versus controls (n = 4). (F) Percentage of Ki67+ cells/gland in Rspo3-WT (n = 5) versus Rspo3-KO mice (n = 5) on day 1 after HDT-induced injury. (G–L) LGR5DTR mice were treated with (G–I) PBS or (J–L) DT for 3 days and sacrificed 24 hours after treatment. Immunofluorescence labeling for (G and J) GIF (green), GSII (red), Ki67 (yellow), (H and K) Ki67, and (I and L) GIF and GSII on sections from PBS-treated mice. (M–O) LGR5DTR mice were treated with DT for 3 days and sacrificed 3 days after treatment. Immunofluorescence labeling for (M) GIF (green), GSII (red), Ki67 (yellow), (N) Ki67, and (O) GIF and GSII. (P and Q) qPCR for (P) Rspo3 or (Q) GIF expression of DT-treated LGR5DTR mice sacrificed on day 1 (n = 5), 3 (n = 4), or 7 (n = 4 mice) versus controls (n = 4). (R–W) Mice were treated with tamoxifen and DT 7 days before euthanasia. (R and U) EGFP expression of agarose sections from (R) LGR5-DTR/EGFP;MYH11-CreERT2;RspoWT/WT and (U) LGR5-DTR/EGFP;MYH11-CreERT2;Rspo3fl/fl mice. (S, T, V, and W) Representative images of sections from (S and T) LGR5-DTR/EGFP;MYH11-CreERT2;RspoWT/WT and (V and W) LGR5-DTR/EGFP;MYH11-CreERT2;Rspo3fl/ft mice stained for (S and V) Ki67 (yellow) and (T and W) GIF (green) and GSII (red). Scale bars: 100 μm. Enlargements in I, L, O, T, and W equal 2:1 magnification. Unpaired parametric t test (F), 1-way ANOVA with Tukey’s multiple-comparison test (C, P, and Q).