Macrophages Promote Progression of Spasmolytic Polypeptide-Expressing Metaplasia After Acute Loss of Parietal Cells

doi:10.1053/j.gastro.2014.02.007

Gastroenterology

Volume 146, Issue 7, June 2014, Pages 1727-1738.e8

https://doi.org/10.1053/j.gastro.2014.02.007 Get rights and content

Background & Aims

Loss of parietal cells causes the development of spasmolytic polypeptide-expressing metaplasia (SPEM) through transdifferentiation of chief cells. In the presence of inflammation, SPEM can advance into a more proliferative metaplasia with increased expression of intestine-specific transcripts. We used L635 to induce acute SPEM with inflammation in mice and investigated the roles of inflammatory cells in the development of SPEM.

Methods

To study the adaptive immune system, Rag1 knockout, interferon-γ–deficient, and wild-type (control) mice received L635 for 3 days. To study the innate immune system, macrophages were depleted by intraperitoneal injection of clodronate liposomes 2 days before and throughout L635 administration. Neutrophils were depleted by intraperitoneal injection of an antibody against Ly6G 2 days before and throughout L635 administration. Pathology and immunohistochemical analyses were used to determine depletion efficiency, metaplasia, and proliferation. To characterize SPEM in each model, gastric tissues were collected and levels of Cftr, Dmbt1, and Gpx2 mRNAs were measured. Markers of macrophage polarization were used to identify subpopulations of macrophages recruited to the gastric mucosa.

Results

Administration of L635 to Rag1 knockout, interferon-γ–deficient, and neutrophil-depleted mice led to development of proliferative SPEM and up-regulation of intestine-specific transcripts in SPEM cells, similar to controls. However, macrophage-depleted mice given L635 showed significant reductions in numbers of SPEM cells, SPEM cell proliferation, and expression of intestine-specific transcripts, compared with control mice given L635. In mice given L635, as well as patients with intestinal metaplasia, M2 macrophages were the primary inflammatory component.

Conclusions

Results from studies of mouse models and human metaplastic tissues indicate that M2 macrophages promote the advancement of SPEM in the presence of inflammation.

Section snippets

L635 treatment

Each experimental group consisted of 3 male mice. L635 (synthesized by the Chemical Synthesis Core of the Vanderbilt Institute of Chemical Biology), dissolved in deionized DNA and RNA-free water, was administered by oral gavage (350 mg/kg) once a day for 3 consecutive days. Neutrophils were depleted through intraperitoneal injection of anti-Ly6G antibody (100 μg, Leaf; BioLegend, San Diego, CA) 2 days before and throughout the 3-day L635 administration. Control mice received intraperitoneal

Rag1 and Interferon-γ Knockout Mice Develop Acute Proliferative Spasmolytic Polypeptide-Expressing Metaplasia

To determine the role of the adaptive immune system in the development of proliferative SPEM, wild-type, Rag1KO, and IFNγKO mice were administered L635 for 3 days and stomach cell lineages were analyzed. Histologic examination revealed parietal cell loss and significant inflammatory infiltration in the fundus of the stomach in all L635-treated mice (Supplementary Figure 1). Upon L635 treatment, neutrophils increased by 6-fold in wild-type mice, with no significant difference observed in

Discussion

The loss of parietal cells, also known as oxyntic atrophy, is considered an absolute requirement for the development of both metaplasia and intestinal-type gastric cancer.7, 8, 9 The exact cause of parietal cell loss during chronic Helicobacter infection remains unclear. Previous studies have investigated the role of immune factors during Helicobacter infection and their impact on parietal cell loss and the development of metaplasia. Severe combined immune-deficient mice do not undergo parietal

Acknowledgments

The authors thank Drs Rupesh Chaturvedi and Keith Wilson for helpful discussions.

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: Conflicts of interest The authors disclose no conflicts.

: Funding These studies were supported by grants from a Department of Veterans Affairs Merit Review Award and NIH RO1 DK071590, as well as a grant from the Martell Foundation (to J.R.G). This work was supported by core resources of the Vanderbilt Digestive Disease Center, (P30 DK058404) the Vanderbilt-Ingram Cancer Center (P30 CA68485, Chemical Synthesis Core), and imaging supported by both the Vanderbilt Combined Imaging Shared Resource and the Vanderbilt Digital Histology Shared Resource.

: Author names in bold designate shared co-first authorship.

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Original ResearchFull Report: Basic and Translational—Alimentary TractMacrophages Promote Progression of Spasmolytic Polypeptide-Expressing Metaplasia After Acute Loss of Parietal Cells

Background & Aims

Methods

Results

Conclusions

Section snippets

L635 treatment

Rag1 and Interferon-γ Knockout Mice Develop Acute Proliferative Spasmolytic Polypeptide-Expressing Metaplasia

Discussion

Acknowledgments

Lancet Oncol

Gastroenterology

Lab Invest

Gastroenterology

Best Pract Res Clin Gastroenterol

Gastroenterology

Gastroenterology

Gastroenterology

Lab Invest

Kidney Int

Am J Pathol

Atherosclerosis

J Biol Chem

Cell

Lab Invest

Gastroenterology

Am J Pathol

Gastroenterology

Immunity

Cancer Cell

Gastroenterology

Parasitism by the “slow” bacterium Helicobacter pylori leads to altered gastric homeostasis and neoplasia

J Clin Invest

Heterogeneity in mouse spasmolytic polypeptide-expressing metaplasia lineages identifies markers of metaplastic progression

Gut

Spasmolytic polypeptide-expressing metaplasia and intestinal metaplasia: time for reevaluation of metaplasias and the origins of gastric cancer

Gastroenterology

Patterns of gastric atrophy in intestinal type gastric carcinoma

Cancer

Alterations in gastric mucosal lineages induced by acute oxyntic atrophy in wild-type and gastrin-deficient mice

Am J Physiol Gastrointest Liver Physiol

Original Research
Full Report: Basic and Translational—Alimentary Tract
Macrophages Promote Progression of Spasmolytic Polypeptide-Expressing Metaplasia After Acute Loss of Parietal Cells