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CD44v9 expression and epithelial proliferation in response to gastric injury. Immunofluorescence staining of CD44v9 (green) and BrdU (red) at the ulcer margin of (A-D) CD44KO, (E-H) BL6, and (I-L) CD44KO BL6 mice 1, 3, 5, and 7 days post-injury. (M) Quantification of CD44v9+/BrdU+ cells per gland from gastric ulcers 1, 3, 5, and 7 days post-injury from CD44KO, BL6, and CD44KO BL6 mice. (N) Quantification of BrdU+/CD44v9− cells per gland from gastric ulcers 1, 3, 5, and 7 days post-injury from CD44KO, BL6, and CD44KO BL6 mice. *p < 0.05 compared with BL6 group, n = 4. Scale bar = 50 μm.
Source publication
Cluster-of-differentiation gene 44, in particular CD44 variant isoform 9 (CD44v9), emerges during regeneration of the gastric epithelium in response to injury. In particular, CD44v9 is expressed within Spasmolytic Polypeptide/TFF2-Expressing Metaplasia (SPEM) glands during gastric repair, but the function is unknown. Here we tested the hypothesis t...
Citations
... Notably, CD44v6 delineates a marker indicative of invasive intramucosal carcinoma and precancerous manifestations. Concurrently, CD44v9 surfaces during gastric epithelial repair post-injury, co-expressing alongside other SPEM-associated markers (55,56). Cumulative evidence underscores the affirmative correlation between CD44 and its variant isoforms with GC onset and progression, pivotal for diagnostic, therapeutic, and prognostic stratifications (57). ...
Gastric cancer (GC) ranks among the top five most diagnosed cancers globally, with particularly high incidence and mortality rates observed in Asian regions. Despite certain advancements achieved through early screening and treatment strategies in many countries, GC continues to pose a significant public health challenge. Approximately 20% of patients infected with Helicobacter pylori develop precancerous lesions, among which metaplasia is the most critical. Except for intestinal metaplasia (IM), which is characterized by goblet cells appearing in the stomach glands, one type of mucous cell metaplasia, spasmolytic polypeptide-expressing metaplasia (SPEM), has attracted much attention. SPEM represents a specific epithelial cell alteration within the gastric mucosa, characterized by the expressing trefoil factor 2 (TFF2) in basal glands, resembling the basal metaplasia of deep antral gland cells. It primarily arises from the transdifferentiation of mature chief cells, mucous neck cells (MNCs), or isthmus stem cells. SPEM is commonly regarded as a precursor lesion in the development of gastric inflammation and subsequent carcinogenesis. The formation of SPEM is intricately associated with chronic gastric inflammation, Helicobacter pylori infection, and various other environmental and genetic factors. Recently, with the profound exploration of the biological and molecular mechanisms underlying SPEM, a deeper understanding of its role in GC initiation and progression has emerged. This review summarizes the role, molecular mechanisms, and clinical significance of SPEM in the onset and progression of GC.
... Moreover, the present study demonstrated that CD44v9 + epithelial cells could be scattered or clustered in normal tissues. Similar studies reported that the CD44v9 expression could occur in the regenerated gastric epithelium cells or gastric epithelium cells infected with H. pylori (36)(37)(38), as well as the expansion of stem cells of gastric tissues (37). ...
The specificity and sensitivity of the current diagnostic and prognostic biomarkers for gastric cancer (GC) are limited. The present study aimed to evaluate the diagnostic and prognostic significance of cluster-of-differentiation gene 44 variant isoform 9 (CD44v9) and T cell immunoglobulin and mucin domain-containing protein 3 (TIM3) expression levels alone or combined in the tumor tissues of patients with GC and reveal the roles of CD44v9 and TIM3 in the cytokeratin (CK)⁺ and CK⁻ regions. Multiplex immunofluorescence staining was performed for CD44v9, TIM3 and CK using a tissue microarray. The tissues were divided into three regions based on CK expression: Total, CK⁺, and CK⁻ regions. The diagnostic and prognostic value was evaluated using receiver operating characteristic curves, Kaplan-Meier and Cox regression analyses. The results demonstrated that the density of cells expressing CD44v9, TIM3 and co-expressing CD44v9 and TIM3 (CD44v9/TIM3) in both the CK⁺ and CK⁻ regions of tumor tissues was significantly higher than those in normal tissues (P<0.001). Moreover, the expression of CD44v9 in the CK⁻ region was significantly positively correlated with age and tumor grade (P<0.05), and the expression of CD44v9/TIM3 in the CK⁻ region of tumor tissues was significantly positively correlated with age, tumor grade and metastasis (P<0.05). Furthermore, the area under the curve for TIM3 expression in the CK⁺ region was 0.709, with a sensitivity of 45.83% and a specificity of 85.54% (P<0.001). High expression of CD44v9 in the CK⁻ region was also significantly associated with poor survival and independently predicted a poor prognosis in patients with GC (hazard ratio, 2.387; 95% confidence interval, 1.384–4.118; P<0.01). In conclusion, dividing tissue regions based on CK expression is important for the diagnosis of GC. The expression of TIM3 in the CK⁺ region demonstrated diagnostic potential for GC, and high expression of CD44v9 in the CK⁻ region was an independent prognostic risk factor for patients with GC.
... mesenchyme(Lee et al, 2023), basement membrane(Bertaux-Skeirik et al, 2017; Khurana et al, 2013), and the immune system(Bockerstett et al, 2020;De Salvo et al, 2021; Jeong et al, 2021;Meyer et al, 2020;Petersen et al, 2018;Petersen et al, 2014). Unfortunately, we are currently limited in our study of the dynamic trafficking changes in cathartocytosis in living cells because we must work in fixed tissue. ...
Injury can cause differentiated cells to undergo massive reprogramming to become proliferative to repair tissue via a cellular program called paligenosis. Gastric digestive-enzyme-secreting chief cells use paligenosis to reprogram into progenitor-like Spasmolytic-Polypeptide Expressing Metaplasia (SPEM) cells. Stage 1 of paligenosis is to downscale mature cell architecture via a process involving lysosomes. Here, we noticed that sulfated glycoproteins (which are metaplasia and cancer markers in mice and humans) were not digested during paligenosis but excreted into the gland lumen. Various genetic and pharmacological approaches showed that endoplasmic reticulum membranes and secretory granule cargo were also excreted and that the process proceeded in parallel with, but was independent lysosomal activity. 3-dimensional light and electron-microscopy demonstrated that excretion occurred via unique, complex, multi-chambered invaginations of the apical plasma membrane. As this lysosome-independent cell cleansing process does not seem to have been priorly described, we termed it "cathartocytosis". Cathartocytosis allows a cell to rapidly eject excess material (likely in times of extreme stress such as are induced by paligenosis) without waiting for autophagic and lysosomal digestion. We speculate the ejection of sulfated glycoproteins (likely mucins) would aid in downscaling and might also help bind and flush pathogens (like H pylori which causes SPEM) away from tissue.
... SPEM lineages have been further characterized as diastase-periodic acid Schiff-positive lineages expressing TFF2 and MUC6 and promoting GSII-agglutinin bonds, similar to mucus-secreting cells of the deep antral glands (12). Furthermore, this lineage expresses several other anal genealogical markers, including CD44 variant 9 (CD44v9) and clusterin, where CD44v9 marks the SPEM in the corpus and is involved in regeneration after gastric epithelial cell injury (7,13,14). These studies have shown a high degree of similarity between the spectra of SPEM and the deep antral glands. ...
Gastric cancer ranks as one of the most prevalent cancers worldwide. While the incidence of gastric cancer in Western countries has notably diminished over the past century, it continues to be a leading cause of cancer-related mortality on a global scale. The majority of gastric cancers in humans are attributed to chronic Helicobacter pylori infection and the progression of gastric cancer is often preceded by gastritis, atrophy, metaplasia and dysplasia. However, the precise mechanisms underlying the development of gastric cancer remain ambiguous, including the formation of gastric polyps and precancerous lesions. In humans, two types of precancerous metaplasia have been identified in relation to gastric malignancies: Intestinal metaplasia and spasmolytic polypeptide-expressing metaplasia (SPEM). The role of SPEM in the induction of gastric cancer has gained recent attention and its link with early-stage human gastric cancer is increasingly evident. To gain insight into SPEM, the present study reviewed the role and research progress of SPEM in gastric cancer.
... The CD44v9positive group has a higher recurrence rate than the CD44v9-negative group [35]. Although CD44v9 emerges in response to injury and contributes to the gastric epithelium [36], there are no reports of CD44v9 directly causing GMA. Further functional studies of rs58618380 in CAPZA1 and CD44v9 are required. ...
Background
Helicobacter pylori secretes cytotoxin-associated gene A (CagA) into the gastric epithelium, causing gastric mucosal atrophy (GMA) and gastric cancer. In contrast, host cells degrade CagA via autophagy. However, the association between polymorphisms in autophagy-related genes and GMA must be fully elucidated.
Results
We evaluated the association between single nucleotide polymorphisms (SNPs) in autophagy-related genes (low-density lipoprotein receptor-related protein 1, LRP1 ; capping actin protein of muscle Z-line alpha subunit 1, CAPAZ1 ; and lysosomal-associated membrane protein 1, LAMP1 ) and GMA in 200 H. pylori -positive individuals. The frequency of the T/T genotype at rs1800137 in LRP1 was significantly lower in the GMA group than in the non-GMA group ( p = 0.018, odds ratio [OR] = 0.188). The frequencies of the G/A or A/A genotype at rs4423118 and T/A or A/A genotype at rs58618380 of CAPAZ1 in the GMA group were significantly higher than those in the non-GMA group ( p = 0.029 and p = 0.027, respectively). Multivariate analysis revealed that C/C or C/T genotype at rs1800137, T/A or A/A genotype at rs58618380, and age were independent risk factors for GMA ( p = 0.038, p = 0.023, and p = 0.006, respectively). Furthermore, individuals with the rs1800137 C/C or C/T genotype of LRP1 had a 5.3-fold higher susceptibility to GMA. These genetic tests may provide future directions for precision medicine for individuals more likely to develop GMA.
Conclusion
LRP1 and CAPZA1 polymorphisms may be associated with the development of GMA.
... As a follow-up to the above studies, Kuo et al. examined the association between serum TFF2 levels and the expression of miR-21, 155 and 223 in gastric mucosa for SPEM and reported that the above molecules might have diagnostic values Kuo et al., 2019). Subsequent studies successively showed that GSII (Shimizu et al., 2016), CD44v9 (Bertaux-Skeirik et al., 2017;Zavros, 2017), Clusterin (Vange et al., 2017), SRY-related high mobility group box gene 9 (SOX9) (Serizawa et al., 2016), human epididymis protein 4 (HE4) (Nozaki et al., 2008;Jeong et al., 2021) and myelin and lymphocyte protein 2 (MAL2) (Weis et al., 2014) were associated with the expression and proliferation of SPEM cells. In recent studies, aquaporin 5 (AQP5), Trop2 and DDIT4 were shown to reflect parietal cell loss and the severity of SPEM development and could also predict a higher risk of GC (Riera et al., 2020;Lee et al., 2021;Miao et al., 2021). ...
Background
Spasmolytic polypeptide expression metaplasia (SPEM) occurs in the corpus of the stomach and is closely related to inflammations caused by H. pylori infection. Recently, SPEM was suggested as one of the dubious precancerous lesions of gastric cancer (GC). Thus, further research on SPEM cell transdifferentiation and its underlying mechanisms could facilitate the development of new molecular targets improving the therapeutics of GC. Using bibliometrics, we analyzed publications, summarized the research hotspots and provided references for scientific researchers engaged in related research fields.
Methods
We searched the Web of Science Core Collection (WoSCC) for publications related to SPEM-GC from 2002 to 2022. The VOSviewer, SCImago, CiteSpace and R software were used to visualize and analyze the data. Gene targets identified in the keyword list were analyzed for functional enrichment using the KEGG and GO databases.
Results
Of the 292 articles identified in the initial search, we observed a stable trend in SPEM-GC research but rapid growth in the number of citations. The United States was the leader in terms of quality publications and international cooperation among them. The total number of articles published by Chinese scholars was second to the United States. Additionally, despite its low centrality and average citation frequency, China has become one of the world’s most dynamic countries in academics. In terms of productivity, Vanderbilt University was identified as the most productive institution. Further, we also observed that Gastroenterology was the highest co-cited journal, and Goldenring Jr. was the most prolific author with the largest centrality.
Conclusion
SPEM could serve as an initial step in diagnosing gastric precancerous lesions. Current hotspots and frontiers of research include SPEM cell lineage differentiation, interaction with H. pylori, disturbances of the mucosal microenvironment, biomarkers, clinical diagnosis and outcomes of SPEM, as well as the development of proliferative SPEM animal models. However, further research and collaboration are still required. The findings presented in this study can be used as reference for the research status of SPEM-GC and determine new directions for future studies.
... These data further support that macrophages are recruited to the site of gastric ulcer and that Hh signaling is needed for the recruitment of macrophages to the site of gastric injury. SmoKO mice exhibit loss of CD44v9 and IL-13 at the site of injury The induction of metaplasia has been shown to correlate with the infiltration of M2 macrophages and a cytokine signaling network of IL-33 and IL-13 15,16 . In particular, our group has demonstrated that the infiltration of macrophages to the gastric epithelium during bacterial infection and regeneration is dependent on Hh signaling 7,14 . ...
... In particular, our group has demonstrated that the infiltration of macrophages to the gastric epithelium during bacterial infection and regeneration is dependent on Hh signaling 7,14 . In addition, CD44v9 marks a reparative cell lineage at the ulcer margin 16 . A loss of CD44v9 at the ulcer margin in SmoKO mice (Fig. 3B) compared to controls (Fig. 3A). ...
... Loss of Shh signaling in the myeloid cell lineage observed in the LysMcre/+;Smof/f (SmoKO) mice correlated with not only a loss of macrophage recruitment to the injured stomach but also disruption of epithelial cell regeneration. Induction of spasmolytic polypeptide/TFF2-expressing metaplasia (SPEM) is critical to driving regeneration of the gastric epithelium 16,30 . Cluster-ofdifferentiation gene 44 variant isoform 9 (CD44v9) emerges during regeneration of the gastric epithelium in response to injury and is known to drive SPEM glands. ...
Sonic Hedgehog (Shh), secreted from gastric parietal cells, contributes to the regeneration of the epithelium. The recruitment of macrophages plays a central role in the regenerative process. The mechanism that regulates macrophage recruitment in response to gastric injury is largely unknown. Here we tested the hypothesis that Shh stimulates macrophage chemotaxis to the injured epithelium and contributes to gastric regeneration. A mouse model expressing a myeloid cell-specific deletion of Smoothened (LysMcre/+;Smof/f) was generated using transgenic mice bearing loxP sites flanking the Smo gene (Smo loxP) and mice expressing a Cre recombinase transgene from the Lysozyme M locus (LysMCre). Acetic acid injury was induced in the stomachs of both control and LysMcre/+;Smof/f (SmoKO) mice and gastric epithelial regeneration and macrophage recruitment analyzed over a period of 7 days post-injury. Bone marrow-derived macrophages (BM-Mø) were collected from control and SmoKO mice. Human-derived gastric organoid/macrophage co-cultures were established, and macrophage chemotaxis measured. Compared to control mice, SmoKO animals exhibited inhibition of ulcer repair and normal epithelial regeneration, which correlated with decreased macrophage infiltration at the site of injury. Bone marrow chimera experiments using SmoKO donor cells showed that control chimera mice transplanted with SmoKO bone marrow donor cells exhibited a loss of ulcer repair, and transplantation of control bone marrow donor cells to SmoKO mice rescued epithelial cell regeneration. Histamine-stimulated Shh secretion in human organoid/macrophage co-cultures resulted in macrophage migration toward the gastric epithelium, a response that was blocked with Smo inhibitor Vismodegib. Shh-induced macrophage migration was mediated by AKT signaling. In conclusion, Shh signaling acts as a macrophage chemoattractant via a Smo-dependent mechanism during gastric epithelial regeneration in response to injury.
... Indeed, certain genes upregulated in metaplastic corpus epithelium are highly expressed in antral glands at homeostasis, including DMBT1 (Sousa et al., 2012;Garay et al., 2017) and GKN3 (Menheniott et al., 2010;Bockerstett et al., 2020), but by the same token, various genes appear to be unique to metaplasia in the corpus, neither expressed in corpus nor antral glands at homeostasis. These include genes such as HE4 (Jeong et al., 2021;Nozaki et al., 2008;O'Neal et al., 2013), CD44v9 (Fan et al., 1996;Bertaux-Skeirik et al., 2015;Bertaux-Skeirik et al., 2017;Tsugawa et al., 2019), and CLU (Weis et al., 2013;Shimizu et al., 2016), among others (Weis et al., 2014). A more appropriate term to describe these glandular changes (and which will be used in this review to refer to these metaplastic changes) is spasmolytic polypeptide-expressing metaplasia (or SPEM), which is characterized by the expression of the antral spasmolytic polypeptide in the bases of corpus glands (Schmidt et al., 1999) and accurately describes the characteristic features of this metaplastic entity. ...
The human stomach functions as both a digestive and innate immune organ. Its main product, acid, rapidly breaks down ingested products and equally serves as a highly effective microbial filter. The gastric epithelium has evolved mechanisms to appropriately handle the myriad of injurious substances, both exogenous and endogenous, to maintain the epithelial barrier and restore homeostasis. The most significant chronic insult that the stomach must face is Helicobacter pylori (Hp), a stomach-adapted bacterium that can colonize the stomach and induce chronic inflammatory and pre-neoplastic changes. The progression from chronic inflammation to dysplasia relies on the decades-long interplay between this oncobacterium and its gastric host. This review summarizes the functional and molecular regionalization of the stomach at homeostasis and details how chronic inflammation can lead to characteristic alterations in these developmental demarcations, both at the topographic and glandular levels. More importantly, this review illustrates our current understanding of the epithelial mechanisms that underlie the pre-malignant gastric landscape, how Hp adapts to and exploits these changes, and the clinical implications of identifying these changes in order to stratify patients at risk of developing gastric cancer, a leading cause of cancer-related deaths worldwide.
... SPEM can be identified by the expression of a number of proteins not expressed in healthy epithelium (CD44v9, GKN3). 20,21,45 As previously reported, SPEM staining was present throughout the corpus of TxA23 mice. 20 In contrast, there was little to no SPEM observed in age-matched TxA23xIl4ra -/mice ( Figure 5B). ...
Background aims:
It is well established that chronic inflammation promotes gastric cancer-associated metaplasia, but little is known regarding the mechanisms by which immune cells and cytokines regulate metaplastic cellular changes. The goals of this study were to identify interleukin 13 (IL-13)-producing immune cells, determine the gastric epithelial cell response(s) to IL-13, and establish the role(s) of IL-13 in metaplasia development.
Methods:
Experiments utilized an established mouse model of autoimmune gastritis (TxA23), TxA23xIl4ra-/- mice, which develop gastritis but do not express the IL-4/IL-13 receptor subunit IL-4Rα, and TxA23xIL-13-YFP mice, which express yellow fluorescent protein in IL-13-producing cells. Flow cytometry was used to measure IL-13 secretion and identify IL-13-producing immune cells. Mouse and human gastric organoids were cultured with IL-13 to determine epithelial cell response(s) to IL-13. Single-cell RNA sequencing was performed on gastric epithelial cells from healthy and inflamed mouse stomachs. Mice with gastritis were administered IL-13-neutralizing antibodies and stomachs were analyzed by histopathology and immunofluorescence.
Results:
We identified six unique subsets of IL-13-producing immune cells in the inflamed stomach. Organoid cultures showed that IL-13 acts directly on gastric epithelium to induce a metaplastic phenotype. IL-4Rα-deficient mice did not progress to metaplasia. Single-cell RNA sequencing determined that gastric epithelial cells from IL-4Rα-deficient mice upregulated inflammatory genes but failed to upregulate metaplasia-associated transcripts. Neutralization of IL-13 significantly reduced and reversed metaplasia development in mice with gastritis.
Conclusions:
IL-13 is made by a variety of immune cell subsets during chronic gastritis and promotes gastric cancer-associated metaplastic epithelial cell changes. Neutralization of IL-13 reduces metaplasia severity during chronic gastritis.
... In contrast to unaffected gastric glands showing no expressions of GSII, CD44v9, or AQP5 at the base, regenerating glands adjacent to ulcerated mucosa prominently showed CD44v9 expression at the base as described previously ( Figure 4A). 27 Most of the CD44v9expressing basal gland cells strongly expressed AQP5 and GSII ( Figure 4A). These findings indicate that AQP5 is expressed in SPEM lineages as part of the regenerative mucosa surrounding acute ulcers. ...
BACKGROUND & AIMS
Metaplasia in the stomach is highly associated with development of intestinal-type gastric cancer. Two types of metaplasias, spasmolytic polypeptide-expressing metaplasia (SPEM) and intestinal metaplasia (IM) are considered precancerous lesions. However, it remains unclear how SPEM and IM are related. Here, we investigated a new lineage-specific marker for SPEM cells, aquaporin 5 (AQP5), to assist in the identification of these two metaplasias.
METHODS
Drug- or Helicobacter felis (H. felis) infection-induced mouse models were used to identify the expression pattern of AQP5 in acute or chronic SPEM. Gene-manipulated mice treated with or without drug were employed to investigate how AQP5 expression is regulated in metaplastic lesions. Metaplastic samples from transgenic mice and human gastric cancer patients were evaluated for AQP5 expression. Immunostaining with lineage-specific markers was used to differentiate metaplastic gland characteristics.
RESULTS
Our results revealed that AQP5 is a novel lineage-specific marker for SPEM cells which are localized at the base of metaplastic glands initially and expand to dominate glands after chronic H. felis infection. Additionally, AQP5 expression was upregulated early in chief cell reprogramming and was promoted by interleukin-13 (IL-13). In humans, metaplastic corpus showed highly branched structures with AQP5-positive SPEM. Human SPEM cells strongly expressing AQP5 were present at the bases of incomplete IM glands marked by TROP2, but were absent from complete IM glands.
CONCLUSIONS
AQP5-expressing SPEM cells are present in pyloric metaplasia and TROP2-positive incomplete IM and may be an important component of metaplasia which can predict a higher risk for gastric cancer development.