Role of metaplasia during gastric regeneration

Spasmolytic polypeptide/trefoil factor 2 (TFF2)-expressing metaplasia (SPEM) is a mucous-secreting reparative lineage that emerges at the ulcer margin in response to gastric injury. Under conditions of chronic inflammation with parietal cell loss, SPEM has been found to emerge and evolve into neoplasia. Cluster-of-differentiation gene 44 (CD44) is known to coordinate normal and metaplastic epithelial cell proliferation. In particular, CD44 variant isoform 9 (CD44v9) associates with the cystine-glutamate transporter xCT, stabilizes the protein, and provides defense against reactive oxygen species (ROS). xCT stabilization by CD44v9 leads to defense against ROS by cystine uptake, glutathione (GSH) synthesis, and maintenance of the redox balance within the intracellular environment. Furthermore, p38 signaling is a known downstream ROS target, leading to diminished cell proliferation and migration, two vital processes of gastric epithelial repair. CD44v9 emerges during repair of the gastric epithelium after injury, where it is coexpressed with other markers of SPEM. The regulatory mechanisms for the emergence of CD44v9 and the role of CD44v9 during the process of gastric epithelial regeneration are largely unknown. Inflammation and M2 macrophage infiltration have recently been demonstrated to play key roles in the induction of SPEM after injury. The following review proposes new insights into the functional role of metaplasia in the process of gastric regeneration in response to ulceration. Our insights are extrapolated from documented studies reporting oxyntic atrophy and SPEM development and our current unpublished findings using the acetic acid-induced gastric injury model.

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Transcriptional Network Analysis Reveals the Role of miR-223-5p During Diabetic Corneal Epithelial Regeneration

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.737472 ◽

2021 ◽

Vol 8 ◽

Author(s):

Yuan Zhang ◽

Shengqian Dou ◽

Xia Qi ◽

Zhenzhen Zhang ◽

Yujie Qiao ◽

...

Keyword(s):

Cell Proliferation ◽

Network Analysis ◽

Nerve Regeneration ◽

Corneal Epithelial ◽

Beneficial Effects ◽

Epithelial Regeneration ◽

Epithelial Wound Healing ◽

Direct Target Gene ◽

Inflammatory Processes

Diabetes mellitus (DM) is a complex metabolic disorder. Long-term hyperglycemia may induce diabetic keratopathy (DK), which is mainly characterized by delayed corneal epithelial regeneration. MicroRNAs (miRNAs) have been reported to play regulatory roles during tissue regeneration. However, the molecular mechanism by which miRNAs influence epithelial regeneration in DK is largely unknown. In this study, we performed miRNA and mRNA sequencing of regenerative corneal epithelium tissue from streptozotocin-induced type 1 diabetic (T1DM) and wild-type mice to screen for differentially expressed miRNAs and mRNAs. Based on regulatory network analysis, miR-223-5p was selected for subsequent experiments and Hpgds was then identified as a direct target gene. MiR-223-5p downregulation significantly promoted diabetic corneal epithelial wound healing and nerve regeneration. However, the beneficial effects of miR-223-5p inhibition were abolished by an Hpgds inhibitor. Furthermore, mechanistic studies demonstrated that miR-223-5p suppression ameliorated inflammation and enhanced cell proliferation signaling in DK. Taken together, our findings revealed that the regulatory role of miR-223-5p in diabetic corneal epithelial and nerve regeneration by mediating inflammatory processes and cell proliferation signaling. And silencing miR-223-5p may contribute to the development of potential therapeutic strategies for DK.

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CD36 maintains the gastric mucosa and associates with gastric disease

Communications Biology ◽

10.1038/s42003-021-02765-z ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Miriam Jacome-Sosa ◽

Zhi-Feng Miao ◽

Vivek S. Peche ◽

Edward F. Morris ◽

Ramkumar Narendran ◽

...

Keyword(s):

Gastric Epithelium ◽

Gastric Injury ◽

Gastric Disease ◽

Tissue Respiration ◽

Gastric Function ◽

Intestinal Hemorrhage ◽

Mitochondrial Efficiency ◽

The Uk ◽

Genetically Determined

AbstractThe gastric epithelium is often exposed to injurious elements and failure of appropriate healing predisposes to ulcers, hemorrhage, and ultimately cancer. We examined the gastric function of CD36, a protein linked to disease and homeostasis. We used the tamoxifen model of gastric injury in mice null for Cd36 (Cd36−/−), with Cd36 deletion in parietal cells (PC-Cd36−/−) or in endothelial cells (EC-Cd36−/−). CD36 expresses on corpus ECs, on PC basolateral membranes, and in gastrin and ghrelin cells. Stomachs of Cd36−/− mice have altered gland organization and secretion, more fibronectin, and inflammation. Tissue respiration and mitochondrial efficiency are reduced. Phospholipids increased and triglycerides decreased. Mucosal repair after injury is impaired in Cd36−/− and EC-Cd36−/−, not in PC-Cd36−/− mice, and is due to defect of progenitor differentiation to PCs, not of progenitor proliferation or mature PC dysfunction. Relevance to humans is explored in the Vanderbilt BioVu using PrediXcan that links genetically-determined gene expression to clinical phenotypes, which associates low CD36 mRNA with gastritis, gastric ulcer, and gastro-intestinal hemorrhage. A CD36 variant predicted to disrupt an enhancer site associates (p < 10−17) to death from gastro-intestinal hemorrhage in the UK Biobank. The findings support role of CD36 in gastric tissue repair, and its deletion associated with chronic diseases that can predispose to malignancy.

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492 TUMOR ASSOCIATED MACROPHAGE PROMOTES EPITHELIAL-TO-MESENCHYMAL TRANSITION IN ESOPHAGEAL SQUAMOUS CELL CANCER

Diseases of the Esophagus ◽

10.1093/dote/doab052.492 ◽

2021 ◽

Vol 34 (Supplement_1) ◽

Author(s):

Ching Tzao ◽

Li-Yuan Cheng ◽

Chien-Chih Chang

Keyword(s):

Cell Proliferation ◽

Epithelial To Mesenchymal Transition ◽

Squamous Cell Cancer ◽

Cell Cancer ◽

M2 Macrophage ◽

Mesenchymal Transition ◽

Tumor Associated Macrophage ◽

Cancer Inflammation ◽

Emt Markers

Abstract We aimed to investigate the role of tumor associated macrophage (TAM) in epithelial-to-mesenchymal transition (EMT) in esophageal squamous cell cancer (ESCC). Methods Expression of CD68 and EMT markers was determined in resected ESCC tumors by immunohistochemistry with clinicopathologic correlation. M2-polarized macrophages were generated from human U937 cells treated with 50 ng/ml phorbol myristate acetate (PMA) while cultured with PMA plus Th2 cytokines. KYSE-510 ESCC cell was co-cultured with M2 macrophages, followed by determination of expression for EMT markers by Western blot. In situ expression of E-cadherin and vimentin was determined using immunofluorescence staining Cell proliferation, invasion and extracellular matrix (ECM) adhesion assays were performed to determine phenotypic characteristics of cultured ESCC cells. Results High expression of CD68 in resected ESCC correlated with worse survival. In addition, expression of CD68 in resected ESCC tumors correlated positively with expression of Snail and vimentin but inversely with E-cadherin. Compared with KYSE-510 cells cultured alone, those co-cultured with M2 macrophage showed higher expression of snail, vimentin, and fibronectin with a more spindle-shaped morphology, suggesting a mesenchymal differentiation. Further, cell proliferation, invasion and ECM adhesion were significantly more pronounced in M2 macrophage co-cultured ESCC cells. Conclusion EMT markers correlated with the number of TAM within resected ESCC tumors, suggesting an association of cancer inflammation in promoting EMT in ESCC. A link between cancer inflammation mediated by TAM deemed to be supported by increased expression of EMT markers and phenotypic changes related to EMT in ESCC cells co-cultured with M2 macrophage. Our results suggest an important role of TAM in promoting EMT in tumor microenvironment with regards to cancer inflammation in ESCC.

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Disruption of selenoprotein biosynthesis affects cell proliferation in the imaginal discs and brain of Drosophila melanogaster

Journal of Cell Science ◽

10.1242/jcs.112.17.2875 ◽

1999 ◽

Vol 112 (17) ◽

pp. 2875-2884

Author(s):

B. Alsina ◽

M. Corominas ◽

M.J. Berry ◽

J. Baguna ◽

F. Serras

Keyword(s):

Cell Proliferation ◽

Drosophila Melanogaster ◽

Mrna Expression ◽

Redox Balance ◽

Imaginal Discs ◽

Brdu Incorporation ◽

Pupal Development ◽

Localization Analysis ◽

Dividing Cells

The patufet gene encodes the Drosophila melanogaster homologue of selenophosphate synthetase, an enzyme required for selenoprotein synthesis, and appears to have a role in cell proliferation. In this paper we analyse the expression pattern of patufet during the development of imaginal discs and brain as well as the function of this gene in relation to cell proliferation. Wild-type organisms showed a highly dynamic pattern of ptuf mRNA expression during larval and pupal development. Co-localization analysis of ptuf mRNA expression and BrdU incorporation showed high levels of ptuf mRNA in dividing cells and low or undetectable levels in non-dividing cells. In addition, [(75)Se] incorporation revealed a major selenoprotein band of 42 kDa. Mutant organisms showed no selenoprotein synthesis, lower levels of cell proliferation, a higher proportion of cells arrested in G(2) as seen by cyclin B labeling and increased levels of reactive oxygen species (ROS). Because most selenoproteins identified so far are antioxidants, the role of ptuf in cell proliferation through the control of the cellular redox balance is discussed.

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Epithelial WNT2B and Desert Hedgehog are necessary for human colonoid regeneration after bacterial cytotoxin injury

10.1101/434639 ◽

2018 ◽

Author(s):

Julie G. In ◽

Jianyi Yin ◽

Michele Doucet ◽

Robert N. Cole ◽

Lauren DeVine ◽

...

Keyword(s):

Cell Proliferation ◽

Stem Cell ◽

Sonic Hedgehog ◽

Hedgehog Signaling ◽

E Coli ◽

Stem Cell Proliferation ◽

Epithelial Regeneration ◽

Desert Hedgehog ◽

Crypt Hyperplasia

SUMMARYIntestinal regeneration and crypt hyperplasia after radiation or pathogen injury relies on Wnt signaling to stimulate stem cell proliferation. Mesenchymal Wnts are essential for homeostasis and regeneration in mice, but the role of epithelial Wnts remains largely uncharacterized. Using the enterohemorrhagicE. colisecreted cytotoxin, EspP to induce injury to human colonoids, we evaluated a simplified, epithelial regeneration model that lacks mesenchymal Wnts. Here, we demonstrate that epithelial-produced WNT2B is upregulated following injury and essential for regeneration. Hedgehog signaling, specifically activation via the ligand Desert Hedgehog (DHH), but not Indian or Sonic Hedgehog, is another driver of regeneration and modulates WNT2B expression. These findings highlight the importance of epithelial WNT2B and DHH in regulating human colonic regeneration after injury.

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4-Hydroxynonenal in Redox Homeostasis of Stomach in Health and Diseases

10.20944/preprints201807.0525.v1 ◽

2018 ◽

Author(s):

Andriy Cherkas ◽

Neven Zarkovic

Keyword(s):

Second Messengers ◽

Redox Homeostasis ◽

Clinical Intervention ◽

Redox Balance ◽

Gastric Epithelium ◽

Gastric Injury ◽

Gastric Diseases ◽

Regeneration Rate ◽

Intervention Trials

The integrity, high functional activity and sufficient regeneration rate of gastric mucosa (GM) in harsh conditions is very challenging pathophysiological demand. The health of gastric epithelium highly depends of efficiency of redox balance maintenance, antioxidant defense and activity of detoxifying systems within the cells as well as robustness of blood supply. Bioactive products of lipid peroxidation, in particular second messengers of free radicals, the bellwether of which is 4-hydroxynonenal (HNE), are important mediators in (patho)physiological adaptive reactions, cells signaling, and are also implicated in pathogenesis of numerous gastric diseases. However, while mechanisms and consequences of HNE and its protein adducts production in response to strong stressors during acute and chronic gastric injury are well studied, many other important issues related to gastric carcinogenesis, tumor growth and progression, the condition of GM after eradication of Helicobacter pylori, the relevance of antioxidants for HNE-related redox homeostasis in GM and many other still need extensive studies and new comprehensive approaches. Therefore, in order to address existing issues preclinical studies and clinical intervention trials are required, which should include also determination of HNE preferably by immunohistological and specific HNE-His ELISA analyses.

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Sonic Hedgehog acts as a macrophage chemoattractant during regeneration of the gastric epithelium

npj Regenerative Medicine ◽

10.1038/s41536-021-00196-2 ◽

2022 ◽

Vol 7 (1) ◽

Author(s):

Jayati Chakrabarti ◽

Martha Dua-Awereh ◽

Michael Schumacher ◽

Amy Engevik ◽

Jennifer Hawkins ◽

...

Keyword(s):

Bone Marrow ◽

Sonic Hedgehog ◽

Gastric Epithelium ◽

Gastric Injury ◽

Macrophage Migration ◽

Epithelial Regeneration ◽

Macrophage Recruitment ◽

Bone Marrow Donor ◽

Donor Cells ◽

Macrophage Chemotaxis

AbstractSonic 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.

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Novel Role of p53 in Septic Immunosuppression: Involvement in Loss and Dysfunction of CD4+ T Lymphocytes

Cellular Physiology and Biochemistry ◽

10.1159/000495241 ◽

2018 ◽

Vol 51 (1) ◽

pp. 452-469 ◽

Cited By ~ 3

Author(s):

Hui Zhang ◽

Cheng-Feng Xu ◽

Chao Ren ◽

Tian-Tian Wu ◽

Ning Dong ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Lymphocytes ◽

Cell Loss ◽

Immune Dysfunction ◽

Jurkat Cells ◽

Cell Counting ◽

Genetic Knockout

Background/Aims: Immunosuppression frequently occurs during the development of sepsis and is closely associated with poor outcome. Characteristics of immunosuppressive CD4+ T lymphocytes in sepsis have been reported to include dramatic cell loss and inactivation. p53 acts as a pivotal transcription factor in regulating cell proliferation and apoptosis, which control tumorigenesis. However, few studies have investigated the universal role of p53 in immune cells, especially in the development of sepsis. Methods: A mouse model of sepsis was produced by cecal ligation and puncture (CLP), and isolated splenic CD4+ T cells or Jurkat cells were exposed to lipopolysaccharide (LPS) stimulation in vitro. We used genetic knockout (p53-/-) mice or the specific inhibitor pifithrin-α (PFT) to investigate the regulatory mechanisms of p53. Cell proliferation ability was assessed using a Cell Counting Kit-8 assay, and apoptotic cells were stained with annexin V/propidium iodide and then analyzed using a FACScan flow cytometer. Protein and mRNA expression levels were measured by western blotting and real-time PCR, and cytokine levels in culture supernatants were determined by enzyme-linked immunosorbent assay. Results: Splenic CD4+ T lymphocytes from CLP mice expressed gradually elevated p53 mRNA and protein levels, which resulted in extracellular regulated protein kinase 1/2 inactivation and expression of apoptotic molecules. Specific inhibition of p53 by PFT or genetic knockout (p53-/-) maintained CD4+ T lymphocyte homeostasis, as indicated by protection from cell loss and restoration of immune function. A medium dose of PFT improved the survival rate of mice, while mortality rate showed only a slight improvement in p53-/- mice compared with wild-type mice. The in vitro responses to LPS were consistent with these results, and upregulation of p53 clearly affected the proliferation, apoptosis, and immune dysfunction of CD4+ T lymphocytes. In addition, we confirmed the regulatory effect of p53 in Jurkat cells, and inhibition of p53 by either inhibition or short hairpin RNA transduction markedly protected cells from LPS stimulation. Conclusion: Elevation of p53 in T lymphocytes during sepsis or endotoxin challenge might be responsible for inhibiting cell proliferation and enhancing both apoptosis and immune dysfunction of T cells.

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The role of NF-κB pathway in cancer inflammation of esophageal squamous carcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.4_suppl.42 ◽

2013 ◽

Vol 31 (4_suppl) ◽

pp. 42-42

Author(s):

Ching Tzao ◽

Li-Yuan Cheng ◽

Chien-Chih Chang ◽

Guang-Huan Sun

Keyword(s):

Cell Proliferation ◽

Tumor Growth ◽

Cell Lines ◽

Angiogenic Factor ◽

Enzyme Linked Immunosorbent Assay ◽

M2 Macrophage ◽

Th2 Cytokines ◽

M2 Macrophages ◽

Cancer Inflammation

42 Background: Chronic inflammation plays an important role in tumorigenesis and tumor progression in human cancers. We aim to investigate the role of NF-kB in cancer inflammation of esophageal squamous cell carcinoma (ESCC). Methods: To generate M2-polarized macrophages, cells of human U937 monocyte cell line were treated with phorbol myristate acetate (PMA, 50 ng/ml) for 6 hours, and then cultured with PMA plus Th2 cytokines, IL-4 (20 ng/ml) and IL-13 (20 ng/ml), for another 18 hours. M2 phenotype was verified by flow cytometry and by cytokine profiling using enzyme-linked immunosorbent assay (ELISA). After co-culture with M2 macrophages, transcription nuclear factor-kB (NF-kB) activity was measured using quantitative polymerase chain reaction (Q-PCR), followed by reconfirmation with Western blot analysis for IkBα in KYSE-170 and -510 ESCC cell lines (kindly provided by Dr. Yutaka Shimada at Kyoto University, Japan). A selective inhibitor to NF-kB, Bay11-7082, was used to treat ESCC cell lines co-cultured with M2 macrophages, followed by cell proliferation, migration, invasion assays and vascular endothelial growth factor (VEGF) secretion by ELISA. The effect of Bay11-7082 (5 mg/kg) against growth of ESCC tumor was tested in xenografted tumors. Results: PMA plus Th2 cytokines treatment promoted differentiation of U937 cells into M2 macrophages. When treated with Bay11-7082, proliferation, migration, invasion and induction of VEGF expression was significantly inhibited in M2 macrophage co-cultured ESCC cells with a down-regulation of IkBα expression. Tumor growth was significantly increased in M2 macrophage co-cultured ESCC cells compared to that of the non-co-cultured controls, which was significantly retarded by treatment with Bay11-7082. Conclusions: NF-kB pathway was activated in ESCC cell lines co-cultured with M2 macrophages with an increase in cell proliferation, cell motility and angiogenic factor in vitro and tumor growth in vivo, which were significantly suppressed by a NF-kB inhibitor, Bay11-7082. These results suggest a role of M2 macrophage in promoting aggressiveness of ESCC cells, possibly through an activation of NF-kB pathway that may serve as a potential therapeutic target for ESCC.

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