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Nature Communications Feb 2021Genetic factors are recognized to contribute to peptic ulcer disease (PUD) and other gastrointestinal diseases, such as gastro-oesophageal reflux disease (GORD),...
Genetic factors are recognized to contribute to peptic ulcer disease (PUD) and other gastrointestinal diseases, such as gastro-oesophageal reflux disease (GORD), irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD). Here, genome-wide association study (GWAS) analyses based on 456,327 UK Biobank (UKB) individuals identify 8 independent and significant loci for PUD at, or near, genes MUC1, MUC6, FUT2, PSCA, ABO, CDX2, GAST and CCKBR. There are previously established roles in susceptibility to Helicobacter pylori infection, response to counteract infection-related damage, gastric acid secretion or gastrointestinal motility for these genes. Only two associations have been previously reported for duodenal ulcer, here replicated trans-ancestrally. The results highlight the role of host genetic susceptibility to infection. Post-GWAS analyses for PUD, GORD, IBS and IBD add insights into relationships between these gastrointestinal diseases and their relationships with depression, a commonly comorbid disorder.
Topics: ABO Blood-Group System; Antigens, Neoplasm; CDX2 Transcription Factor; Depression; Duodenal Ulcer; Female; Fucosyltransferases; GPI-Linked Proteins; Galactosyltransferases; Gastroesophageal Reflux; Gastrointestinal Diseases; Genetic Predisposition to Disease; Genome-Wide Association Study; Helicobacter Infections; Helicobacter pylori; Humans; Inflammatory Bowel Diseases; Male; Mucin-1; Mucin-6; Neoplasm Proteins; Peptic Ulcer; Galactoside 2-alpha-L-fucosyltransferase
PubMed: 33608531
DOI: 10.1038/s41467-021-21280-7 -
Science (New York, N.Y.) Jun 2019Glycosylation alterations are indicative of tissue inflammation and neoplasia, but whether these alterations contribute to disease pathogenesis is largely unknown. To...
Glycosylation alterations are indicative of tissue inflammation and neoplasia, but whether these alterations contribute to disease pathogenesis is largely unknown. To study the role of glycan changes in pancreatic disease, we inducibly expressed human fucosyltransferase 3 and β1,3-galactosyltransferase 5 in mice, reconstituting the glycan sialyl-Lewis, also known as carbohydrate antigen 19-9 (CA19-9). Notably, CA19-9 expression in mice resulted in rapid and severe pancreatitis with hyperactivation of epidermal growth factor receptor (EGFR) signaling. Mechanistically, CA19-9 modification of the matricellular protein fibulin-3 increased its interaction with EGFR, and blockade of fibulin-3, EGFR ligands, or CA19-9 prevented EGFR hyperactivation in organoids. CA19-9-mediated pancreatitis was reversible and could be suppressed with CA19-9 antibodies. CA19-9 also cooperated with the oncogene to produce aggressive pancreatic cancer. These findings implicate CA19-9 in the etiology of pancreatitis and pancreatic cancer and nominate CA19-9 as a therapeutic target.
Topics: Acute Disease; Animals; CA-19-9 Antigen; Carcinogenesis; Carcinoma, Pancreatic Ductal; Cell Line, Tumor; Chronic Disease; ErbB Receptors; Extracellular Matrix Proteins; Fucosyltransferases; Galactosyltransferases; Glycosylation; Humans; Mice; Molecular Targeted Therapy; Pancreatic Neoplasms; Pancreatitis
PubMed: 31221853
DOI: 10.1126/science.aaw3145 -
Nature Communications Jan 2018The communication between tumor-derived elements and stroma in the metastatic niche has a critical role in facilitating cancer metastasis. Yet, the mechanisms tumor...
The communication between tumor-derived elements and stroma in the metastatic niche has a critical role in facilitating cancer metastasis. Yet, the mechanisms tumor cells use to control metastatic niche formation are not fully understood. Here we report that in the lung metastatic niche, high-metastatic hepatocellular carcinoma (HCC) cells exhibit a greater capacity to convert normal fibroblasts to cancer-associated fibroblasts (CAFs) than low-metastatic HCC cells. We show high-metastatic HCC cells secrete exosomal miR-1247-3p that directly targets B4GALT3, leading to activation of β1-integrin-NF-κB signaling in fibroblasts. Activated CAFs further promote cancer progression by secreting pro-inflammatory cytokines, including IL-6 and IL-8. Clinical data show high serum exosomal miR-1247-3p levels correlate with lung metastasis in HCC patients. These results demonstrate intercellular crosstalk between tumor cells and fibroblasts is mediated by tumor-derived exosomes that control lung metastasis of HCC, providing potential targets for prevention and treatment of cancer metastasis.
Topics: Animals; Cancer-Associated Fibroblasts; Carcinoma, Hepatocellular; Cell Communication; Cell Line, Tumor; Cell Transformation, Neoplastic; Exosomes; Gene Expression Regulation, Neoplastic; Humans; Integrin beta1; Interleukin-6; Interleukin-8; Liver Neoplasms; Lung Neoplasms; Male; Mice; Mice, Nude; MicroRNAs; N-Acetyllactosamine Synthase; Neoplasm Invasiveness; Neoplasm Transplantation; Neoplastic Cells, Circulating; Signal Transduction
PubMed: 29335551
DOI: 10.1038/s41467-017-02583-0 -
Cold Spring Harbor Protocols Jun 2015The Golgi apparatus is a membranous organelle that modifies and packages proteins and lipids into transport carriers and sends them to the proper locations in the cell....
The Golgi apparatus is a membranous organelle that modifies and packages proteins and lipids into transport carriers and sends them to the proper locations in the cell. The study of Golgi structure and function can be facilitated by the isolation of this organelle from homogenates of tissues or cells. Liver cells have abundant Golgi membranes because they actively secrete proteins and lipids; therefore, liver tissue is often the preferred source. In this protocol, Golgi membranes are purified from rat liver homogenate by two sequential sucrose gradients. The relative yield of the prepared Golgi stacks is then assessed by measuring the increase in activity of a Golgi marker enzyme, β-1,4-galactosyltransferase, over that of the total liver homogenate. A typical preparation can yield Golgi membranes that are purified 80- to 100-fold over the homogenate, and the majority (60%-70%) retain their stacked nature.
Topics: Animals; Cell Fractionation; Centrifugation, Density Gradient; Female; Golgi Apparatus; Lipids; Liver; N-Acetyllactosamine Synthase; Proteins; Rats; Rats, Sprague-Dawley
PubMed: 26034300
DOI: 10.1101/pdb.prot075911 -
Journal of Experimental & Clinical... Oct 2022Beta-1,3-galactosyltransferase-4 (B3GALT4) plays a critical regulatory role in tumor biology. However, the role of B3GALT4 in modulating the tumor microenvironment (TME)...
BACKGROUND
Beta-1,3-galactosyltransferase-4 (B3GALT4) plays a critical regulatory role in tumor biology. However, the role of B3GALT4 in modulating the tumor microenvironment (TME) of neuroblastoma (NB) remains unknown.
METHODS
Public datasets and clinical NB samples were collected to evaluate the expression and clinical significance of GD2 and B3GALT4 in NB patients. CCK-8, colony formation, and transwell assays and experiments in tumor-bearing mouse models were conducted to investigate the function of B3GALT4. Flow cytometry, ELISA, immunohistochemistry, immunofluorescence, western blotting, and chemotaxis assays were conducted to ascertain the immunomodulatory mechanism of B3GALT4. The combined therapeutic effect of the lipid raft inhibitor MβCD and anti-GD2 mAb was validated in a murine model of NB.
RESULTS
GD2 was overexpressed in NB tissues and high expression of GD2 was associated with poor prognosis in NB patients. B3GALT4 was downregulated in NB tissues, and low expression of B3GALT4 indicated poor prognosis in NB patients. Silencing B3GALT4 significantly enhanced tumor progression both in vitro and in vivo. Meanwhile, the overexpression of B3GALT4 increased the recruitment of CD8 T lymphocytes via the chemokines CXCL9 and CXCL10. Additionally, B3GALT4 regulated NB-cell GD2 expression and lipid raft formation. Mechanistically, B3GALT4 regulated the expression of CXCL9 and CXCL10 via the c-Met signaling in the lipid rafts and the downstream AKT/mTOR/IRF-1 pathway. The lipid raft inhibitor, MβCD, attenuated B3GALT4 deficiency-induced tumor progression and immune evasion. Last, MβCD combined with anti-GD2 mAb treatment significantly enhanced the antitumor effect and the infiltration of CD8 T cells.
CONCLUSIONS
Upregulation of B3GALT4 promotes the secretion of CXCL9 and CXCL10 to recruit CD8 T lymphocytes via the GD2-mediated lipid rafts and the c-Met/AKT/mTOR/IRF-1 pathway. Moreover, lipid raft inhibitors may enhance the efficacy of anti-GD2 immunotherapy for NB.
Topics: Animals; Mice; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Chemokines; Galactosyltransferases; Gangliosides; Membrane Microdomains; Neuroblastoma; Proto-Oncogene Proteins c-akt; Sincalide; TOR Serine-Threonine Kinases; Tumor Microenvironment; Proto-Oncogene Proteins c-met
PubMed: 36284313
DOI: 10.1186/s13046-022-02523-x -
Molecular Cell Jan 2023Post-translational modifications (PTMs) of proteins are crucial to guarantee the proper biological functions in immune responses. Although protein phosphorylation has...
Post-translational modifications (PTMs) of proteins are crucial to guarantee the proper biological functions in immune responses. Although protein phosphorylation has been extensively studied, our current knowledge of protein pyrophosphorylation, which occurs based on phosphorylation, is very limited. Protein pyrophosphorylation is originally considered to be a non-enzymatic process, and its function in immune signaling is unknown. Here, we identify a metabolic enzyme, UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1), as a pyrophosphorylase for protein serine pyrophosphorylation, by catalyzing the pyrophosphorylation of interferon regulatory factor 3 (IRF3) at serine (Ser) 386 to promote robust type I interferon (IFN) responses. Uap1 deficiency significantly impairs the activation of both DNA- and RNA-viruse-induced type I IFN pathways, and the Uap1-deficient mice are highly susceptible to lethal viral infection. Our findings demonstrate the function of protein pyrophosphorylation in the regulation of antiviral responses and provide insights into the crosstalk between metabolism and innate immunity.
Topics: Animals; Mice; Immunity, Innate; Interferon Regulatory Factor-3; Interferon Type I; Phosphorylation; Signal Transduction; Galactosyltransferases
PubMed: 36603579
DOI: 10.1016/j.molcel.2022.12.007 -
Frontiers in Plant Science 2021A wide range of proteins with diverse functions in development, defense, and stress responses are -arabinosylated at hydroxyprolines (Hyps) within distinct amino acid... (Review)
Review
A wide range of proteins with diverse functions in development, defense, and stress responses are -arabinosylated at hydroxyprolines (Hyps) within distinct amino acid motifs of continuous stretches of Hyps, as found in the structural cell wall extensins, or at non-continuous Hyps as, for example, found in small peptide hormones and a variety of plasma membrane proteins involved in signaling. Plant -glycosylation relies on hydroxylation of Prolines to Hyps in the protein backbone, mediated by prolyl-4-hydroxylase (P4H) which is followed by -glycosylation of the Hyp C-OH group by either galactosyltransferases (GalTs) or arabinofuranosyltranferases (AraTs) yielding either Hyp-galactosylation or Hyp-arabinosylation. A subset of the P4H enzymes with putative preference to hydroxylation of continuous prolines and presumably all AraT enzymes needed for synthesis of the substituted arabinose chains of one to four arabinose units, have been identified and functionally characterized. Truncated root-hair phenotype is one common denominator of mutants of Hyp formation and Hyp-arabinosylation glycogenes, which act on diverse groups of -glycosylated proteins, e.g., the small peptide hormones and cell wall extensins. Dissection of different substrate derived effects may not be regularly feasible and thus complicate translation from genotype to phenotype. Recently, lack of proper arabinosylation on arabinosylated proteins has been shown to influence their transport/fate in the secretory pathway, hinting to an additional layer of functionality of -arabinosylation. Here, we provide an update on the prevalence and types of -arabinosylated proteins and the enzymatic machinery responsible for their modifications.
PubMed: 33815452
DOI: 10.3389/fpls.2021.645219 -
Current Opinion in Organ Transplantation Dec 2017This review describes recent progress in tolerance-inducing strategies across xenogeneic immunological barriers as well as the potential benefit of a tolerance strategy... (Review)
Review
PURPOSE OF REVIEW
This review describes recent progress in tolerance-inducing strategies across xenogeneic immunological barriers as well as the potential benefit of a tolerance strategy for islets and kidney xenotransplantation.
RECENT FINDINGS
Using advanced gene editing technologies, xenotransplantation from multitransgenic alpha-1,3-galactosyltransferase knockout pigs has demonstrated marked prolongation of renal xenograft survival, ranging from days to greater than several months for life-supporting kidneys, and more than 2 years in a heterotopic nonlife-supporting cardiac xenograft model. Continuous administration of multiple immunosuppressive drugs has been required and attempts to taper immunosuppression have been unsuccessful. It appears likely that low levels of T cell dependent antibodies and activation of innate responses are responsible for xenograft loss. Mixed chimerism and thymic transplantation approaches have achieved xenogeneic tolerance in pig-to-mouse models and both have recently been extended to pig-to-baboon models. Encouraging results have been reported, including persistence of macrochimerism, prolonged pig skin graft survival, donor-specific unresponsiveness in vitro and detection of recent T cell emigrants in vivo.
SUMMARY
Although tolerance induction in vivo has not yet been achieved in pig-to-baboon models, recent results are encouraging that this goal will be attainable through genetic engineering of porcine donors.
Topics: Animals; Chimerism; Humans; Immune Tolerance; Papio; Swine; Transplantation, Heterologous
PubMed: 28937406
DOI: 10.1097/MOT.0000000000000466 -
Oncology Letters Aug 2021O-linked glycosylation (-glycosylation) and N-linked glycosylation (-glycosylation) are the two most important forms of protein glycosylation, which is an important... (Review)
Review
O-linked glycosylation (-glycosylation) and N-linked glycosylation (-glycosylation) are the two most important forms of protein glycosylation, which is an important post-translational modification. The regulation of protein function involves numerous mechanisms, among which protein glycosylation is one of the most important. Core 1 synthase glycoprotein-N-acetylgalactosamine 3-β-galactosyltransferase 1 () serves an important role in the regulation of -glycosylation and is an essential enzyme for synthesizing the core 1 structure of mucin-type O-glycans. Furthermore, serves a vital role in a number of biological functions, such as angiogenesis, platelet production and kidney development. Impaired expression activity has been associated with different types of human diseases, including inflammatory or immune-mediated diseases, and cancer. -glycosylation exists in normal tissues, as well as in tumor tissues. Previous studies have revealed that changes in the level of glycosyltransferase in different types of cancer may be used as potential therapeutic targets. Currently, numerous studies have reported the dual role of in tumors (carcinogenesis and cancer suppression). The present review reports the role of in normal development and human diseases. Since the mechanism and regulation of and -glycosylation remain elusive, further studies are required to elucidate their effects on development and disease.
PubMed: 34149900
DOI: 10.3892/ol.2021.12850 -
Oncology Letters Mar 2022Cancer remains one of the most difficult diseases to treat. In the quest for early diagnoses to improve patient survival and prognosis, targeted therapies have become a... (Review)
Review
Cancer remains one of the most difficult diseases to treat. In the quest for early diagnoses to improve patient survival and prognosis, targeted therapies have become a hot research topic in recent years. Glycosylation is the most common posttranslational modification in mammalian cells. Core 1β1,3-galactosyltransferase (C1GALT1) is a key glycosyltransferase in the glycosylation process and is the key enzyme in the formation of the core 1 structure on which most complex and branched O-glycans are formed. A recent study reported that C1GALT1 was aberrantly expressed in tumors. In cancer cells, C1GALT1 is regulated by different factors. In the present review, the expression of C1GALT1 in different tumors and its possible molecular mechanisms of action are described and the role of C1GALT1 in cancer development is discussed.
PubMed: 35154428
DOI: 10.3892/ol.2022.13217