g(m1)-ganglioside and Neoplasms

Glycoprotein glycan chains, by virtue of structure, topology of presentation and connection to signal-inducing units, are functional galectin counterreceptors. As example, cross-linking of the α(5)β(1) integrin by galectin-1 on carcinoma cells leads to G(1) arrest or anoikis. Contact-dependent switching from proliferation to differentiation in cultured neuroblastoma cells (SK-N-MC) also utilizes galectin-1. Activity enhancement of a cell surface sialidase underlies the shift in glycan display to ganglioside GM1. Its pentasaccharide within microdomains becomes the target. Similarly, this recognition pair is upregulated upon T cell activation. Cross-linking of GM1 along with associated α(4)/α(5)β(1) integrins elicits Ca(2+)-influx via TRPC5 channels as the relevant response for T effector cell (T(eff)) suppression. Unlike T(eff) cells from wild-type mice, those from genetically altered mice lacking GM1 are not suppressed by galectin-1 or regulatory T cells. Similarly, in the context of GM1 deficiency in NOD mice, T(eff) cells are associated with resistance to regulatory T cell suppression, which is reversed by applied GM1. The broad array of glycosphingolipid structures suggests the possible existence of several novel counterreceptors targeted to endogenous lectins, with sulfatide-galectin-4 interplay within apical delivery serving as recent example.

Topics: Animals; Anoikis; Cell Communication; G(M1) Ganglioside; Galectins; Glycoproteins; Humans; Indazoles; Mice; Models, Immunological; Morpholines; Neoplasms; Propionates; Signal Transduction; T-Lymphocytes

Topics: Animals; Emulsions; G(M1) Ganglioside; Immunotherapy; Mice; N-Acetylneuraminic Acid; Neoplasms; Selectins; Ubiquinone

On-chip glycan biosynthesis is an effective strategy for preparing useful complex glycan sources and for preparing glycan-involved applications simultaneously. However, current methods have some limitations when analyzing biosynthesized glycans and optimizing enzymatic reactions, which could result in undefined glycan structures on a surface, leading to unequal and unreliable results. In this work, a glycan chip is developed by introducing a pH-responsive i-motif DNA linker to control the immobilization and isolation of glycans on chip surfaces in a pH-dependent manner. On-chip enzymatic glycosylations are optimized for uniform biosynthesis of cancer-associated Globo H hexasaccharide and its related complex glycans through stepwise quantitative analyses of isolated products from the surface. Successful interaction analyses of the anti-Globo H antibody and MCF-7 breast cancer cells with on-chip biosynthesized Globo H-related glycans demonstrate the feasibility of the structure-switchable DNA linker-based glycan chip platform for on-chip complex glycan biosynthesis and glycan-involved applications.

Topics: Antigens, Tumor-Associated, Carbohydrate; Cholera Toxin; DNA; G(M1) Ganglioside; Glycosylation; Humans; Hydrogen-Ion Concentration; MCF-7 Cells; Neoplasms; Oligonucleotide Array Sequence Analysis; Polysaccharides; Protein Subunits

SeviL is a recently isolated lectin found to bind to the linear saccharides of the ganglioside GM1b (Neu5Ac[Formula: see text](2-3)Gal[Formula: see text](1-3)GalNAc[Formula: see text](1-4)Gal[Formula: see text](1-4)Glc) and its precursor, asialo-GM1 (Gal[Formula: see text](1-3)GalNAc[Formula: see text](1-4)Gal[Formula: see text](1-4)Glc). The crystal structures of recombinant SeviL have been determined in the presence and absence of ligand. The protein belongs to the [Formula: see text]-trefoil family, but shows only weak sequence similarity to known structures. SeviL forms a dimer in solution, with one binding site per subunit, close to the subunit interface. Molecular details of glycan recognition by SeviL in solution were analysed by ligand- and protein-based NMR techniques as well as ligand binding assays. SeviL shows no interaction with GM1 due to steric hindrance with the sialic acid branch that is absent from GM1b. This unusual specificity makes SeviL of great interest for the detection and control of certain cancer cells, and cells of the immune system, that display asialo-GM1.

Topics: Animals; Bivalvia; Carbohydrate Sequence; G(M1) Ganglioside; Gangliosides; Humans; Lectins; Neoplasms

Topics: Alzheimer Disease; Animals; Blood Platelets; Cell Membrane; Delayed-Action Preparations; Drug Delivery Systems; G(M1) Ganglioside; Humans; Magnets; Nanoparticles; Neoplasms; Neutrophils

Cancer progression involves carcinogenesis, an increase in tumour size, and metastasis. Here, we investigated the effect of overexpressed CXC chemokine ligand 14 (CXCL14) on these processes by using CXCL14/BRAK (CXCL14) transgenic (Tg) mice. The rate of AOM/DSS-induced colorectal carcinogenesis in these mice was significantly lower compared with that for isogenic wild type C57BL/6 (Wt) mice. When tumour cells were injected into these mice, the size of the tumours that developed and the number of metastatic nodules in the lungs of the animals were always significantly lower in the Tg mice than in the Wt ones. Injection of anti-asialo-GM1 antibodies to the mice before and after injection of tumour cells attenuated the suppressing effects of CXCL14 on the tumor growth and metastasis, suggesting that NK cell activity played an important role during CXCL14-mediated suppression of tumour growth and metastasis. The importance of NK cells on the metastasis was also supported when CXCL14 was expressed in B16 melanoma cells. Further, the survival rates after tumour cell injection were significantly increased for the Tg mice. As these Tg mice showed no obvious abnormality, we propose that CXCL14 to be a promising molecular target for cancer suppression/prevention.

Topics: Animals; Antigens, Ly; Autoantibodies; Cell Transformation, Neoplastic; Chemokines, CXC; Chronic Disease; Colitis; Disease Models, Animal; Female; G(M1) Ganglioside; Galactosylceramides; Killer Cells, Natural; Lung Neoplasms; Lymphocyte Depletion; Melanoma, Experimental; Mice; Mice, Transgenic; Neoplasms; NK Cell Lectin-Like Receptor Subfamily B; Tumor Burden

GM1 and GM2 gangliosides are important components of the cell membrane and play an integral role in cell signaling and metabolism. In this conceptual overview, we discuss recent developments in our understanding of the basic biological functions of GM1 and GM2 and their involvement in several diseases. In addition to a well-established spectrum of disorders known as gangliosidoses, such as Tay-Sachs disease, more and more evidence points at an involvement of GM1 in Alzheimer's and Parkinson's diseases. New emerging methodologies spanning from single-molecule imaging in vivo to simulations in silico have complemented standard studies based on ganglioside extraction.

Topics: Amyloid beta-Peptides; Cell Membrane; Diabetes Mellitus; G(M1) Ganglioside; G(M2) Ganglioside; Humans; Neoplasms; Neurodegenerative Diseases; Virus Diseases

Hypoxia-inducible factor 1 alpha (HIF-1alpha), a component of HIF-1, is expressed in human tumors and renders cells able to survive and grow under hypoxic (low-oxygen) conditions. YC-1, 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole, an agent developed for circulatory disorders that inhibits platelet aggregation and vascular contraction, inhibits HIF-1 activity in vitro. We tested whether YC-1 inhibits HIF-1 and tumor growth in vivo.. Hep3B hepatoma, NCI-H87 stomach carcinoma, Caki-1 renal carcinoma, SiHa cervical carcinoma, and SK-N-MC neuroblastoma cells were grown as xenografts in immunodeficient mice (69 mice total). After the tumors were 100-150 mm(3), mice received daily intraperitoneal injections of vehicle or YC-1 (30 microg/g) for 2 weeks. HIF-1 alpha protein levels and vascularity in tumors were assessed by immunohistochemistry, and the expression of HIF-1-inducible genes (vascular endothelial growth factor, aldolase, and enolase) was assessed by reverse transcription-polymerase chain reaction. All statistical tests were two-sided.. Compared with tumors from vehicle-treated mice, tumors from YC-1-treated mice were statistically significantly smaller (P<.01 for all comparisons), expressed lower levels of HIF-1 alpha (P<.01 for all comparisons), were less vascularized (P<.01 for all comparisons), and expressed lower levels of HIF-1-inducible genes, regardless of tumor type.. The inhibition of HIF-1 alpha activity in tumors from YC-1-treated mice is associated with blocked angiogenesis and an inhibition of tumor growth. YC-1 has the potential to become the first antiangiogenic anticancer agent to target HIF-1 alpha.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Carcinoma; Carcinoma, Hepatocellular; Cell Hypoxia; Culture Media, Conditioned; Endothelial Growth Factors; Enzyme-Linked Immunosorbent Assay; Female; G(M1) Ganglioside; Gene Expression Regulation, Neoplastic; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoblotting; Indazoles; Intercellular Signaling Peptides and Proteins; Kidney Neoplasms; Killer Cells, Natural; Liver Neoplasms; Lymphokines; Male; Mice; Mice, SCID; Neoplasms; Neovascularization, Pathologic; Neuroblastoma; Platelet Endothelial Cell Adhesion Molecule-1; Precipitin Tests; Rats; Reverse Transcriptase Polymerase Chain Reaction; Stomach Neoplasms; Transcription Factors; Transplantation, Heterologous; Tumor Cells, Cultured; Uterine Cervical Neoplasms; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

We previously reported that ganglioside GD1a, which is highly expressed in poorly metastatic FBJ-S1 cells, inhibits the serum-induced motility of FBJ-LL cells and that the metastatic potential of FBJ-LL cells is completely suppressed by enforced GD1a expression (Hyuga et al., Int J Cancer 1999;83:685-91). We recently discovered that hepatocyte growth factor (HGF) induces FBJ-LL cell motility. In the present study, the HGF-induced motility of FBJ-S1 cells was found to be one-thirtieth that of FBJ-LL cells. This motility of GD1a-expressing transfectants, which were produced by transfection of FBJ-LL cells with GM2/GD2 synthase cDNA, decreased with increases in their GD1a expression and HGF induced almost no motility in GD1a-pretreated FBJ-LL cells, indicating that GD1a inhibits the HGF-induced motility of FBJ-LL cells. The expression of the HGF receptor c-Met on FBJ-S1 cells, FBJ-LL cells, transfectants and a mock-transfectant was almost the same. The level of tyrosine phosphorylation of c-Met after HGF stimulation in FBJ-S1 cells, GD1a-pretreated FBJ-LL cells and a GD1a-expressing transfectant was significantly lower than in FBJ-LL cells and a mock-transfectant. These findings suggested that GD1a inhibits the HGF-induced motility of FBJ-LL cells through suppression of tyrosine phosphorylation of c-Met. HepG2 cells, a human hepatoma cell line, were used to investigate whether GD1a interferes with other cancer cells expressing c-Met. HepG2 cells did not express GD1a. HGF induced cell scattering of HepG2 cells and the scattering was inhibited by pretreating the cells with GD1a. The c-Met in the cells was autophosphorylated by stimulation with HGF, but after treating the cells with GD1a, the HGF-induced autophosphorylation of c-Met was suppressed. These results suggest that GD1a acts as a negative regulator of c-Met in cancer cells.

Topics: Actins; Animals; Blotting, Western; Cell Movement; DNA, Complementary; Flow Cytometry; G(M1) Ganglioside; Gangliosides; Hepatocyte Growth Factor; Mice; Neoplasms; Phosphorylation; Precipitin Tests; Proto-Oncogene Proteins c-met; Signal Transduction; Stress Fibers; Time Factors; Transfection; Tumor Cells, Cultured; Tyrosine

The complement system is one potential cytotoxic effector mechanism that might be effective in immunotherapy of cancer using monoclonal antibodies (mAb) directed against tumor antigens. In order to evaluate the treatment outcome from trials using mAb in cancer patients, assessment of complement-dependent cytotoxicity (CDC) may therefore be of interest. Here we describe the elaboration of a CDC assay in vitro using a rat hepatoma cell line, H4-II-E, as target cells sensitised with mAb F12, directed against the tumor-associated ganglioside antigen fucosyl-GMI. Sensitised cells were incubated with various concentrations of fresh serum as complement source for 48 h and cytotoxicity was then assessed by the tetrazolium bromide (MTT) test. A large variation in CDC efficacy was observed between individual serum donors. No differences in CDC could be seen between healthy donors and cancer patients. The CDC showed a strong correlation to the serum concentrations of complement factor C4, supporting the validity of the assay. Our results suggest that there may be significant variations in complement function within and between individuals that might influence the outcome of clinical mAb therapy. The H4/F12 CDC assay described here, together with measurement of individual complement factors. such as C4, should be further validated in cancer patients at various disease stages and phases of treatment.

Topics: Animals; Antibodies, Monoclonal; Cell Death; Cell Division; Chemistry, Clinical; Complement C3c; Complement C4; Complement System Proteins; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; G(M1) Ganglioside; Humans; Immunoglobulin G; Mice; Neoplasms; Rats; Tetrazolium Salts; Thiazoles; Time Factors; Tumor Cells, Cultured

This study is intended to demonstrate the versatility and feasibility of custom-made oligosaccharide-exposing neoglycoconjugates including histo-blood group epitopes in various human lesions, including nasal polyps. The binding of the biotinylated probes was determined on formalin-fixed paraffin-embedded sections from archive materials. The general aspects of our results may be interpreted as follows: the neoglycoconjugates used here can readily detect differences in the ability of cells to bind glycan residues in tissue sections, thereby enabling the extent of the binding capacity of various types of human lesions to be compared. Furthermore, the reactivity to glycan may reflect characteristics of the cells and their environment. The investigation into pathological disorders with respect to the binding capacity of these carrier-immobilized mono- or oligosaccharide structures derived from custom-made synthesis or biochemical purification is based on the prospect of translating progress in this field into the establishment of potentially beneficial procedures for medical diagnosis and pathological classification.

Topics: Adenocarcinoma; Binding Sites; Blood Group Antigens; Brain Neoplasms; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Transitional Cell; Colonic Neoplasms; Feasibility Studies; Female; Fibroadenoma; G(M1) Ganglioside; Glioblastoma; Glycoconjugates; Histocytochemistry; Humans; Hyaluronic Acid; Male; Melanoma; Nasal Polyps; Neoplasms; Prostatic Hyperplasia; Skin Neoplasms; Trisaccharides; Urinary Bladder Neoplasms

N-Glycolylglucosamine 8 was synthesized in 4 steps from anisal glucosamine, via the new crystalline monochloracetyl derivatives 3, 4 and 7. N-Glycolylneuraminic acid 10 was prepared in 59% yield starting from pyruvate and a mixture of 8 and its manno epimer 9 in a 2:3 ratio, with immobilized sialic acid aldolase. Neu5Gc 10 was converted into CMP-NeuGc 11 in the presence of immobilized calf brain CMP-sialate synthetase. Finally 11 was used as a donor in the transfer to the acceptor beta-D-Gal-(1-3)-beta-D-GalNAc-OBn 12 catalyzed by a preparation of porcine liver (2-3)-alpha-sialyltransferase, roughly purified by a chromatography on Cibacron Blue-agarose. alpha-Neu5Gc-(2-3)-beta-D-Gal-(1-3)-beta-D-GalNac-OBn 13 isolated in 56% yield was deprotected to give the non-reducing terminal sequence of GM1b glycolylated ganglioside, which might be expressed in human tumors.

Topics: Animals; beta-Galactoside alpha-2,3-Sialyltransferase; Biomarkers, Tumor; Brain; Carbohydrate Conformation; Carbohydrate Sequence; Cattle; Chromatography, Affinity; Enzymes, Immobilized; Fructose-Bisphosphate Aldolase; G(M1) Ganglioside; Humans; Indicators and Reagents; Liver; Magnetic Resonance Spectroscopy; Molecular Sequence Data; N-Acylneuraminate Cytidylyltransferase; Neoplasms; Oligopeptides; Optical Rotation; Sialyltransferases; Swine

We have examined the antitumor and antimetastatic effects of native-type, glycosylated recombinant lymphotoxin (LT) on human and murine tumors transplanted in mice. The results reported here are as follows: (a) The in vivo antitumor spectrum of LT is not coincident with the in vitro study, and it has a wide antitumor spectrum and substantially inhibits the growth of human solid tumors, (b) When both syngeneic and nude mice are transplanted with Meth A tumor, the significant growth-inhibitory effect of LT is obtained in syngeneic mice, but the effect is quite small in nude mice regardless of the routes; LT attains the same degree of effectiveness as that in syngeneic mice, but at an 8 to 16 times higher dose. Furthermore, the pretreatment with anti-asialo-GM1 antibody inhibits the antitumor effects of LT in syngeneic mice, (c) In the pulmonary metastasis model induced by i.v. injection of Meth A cells, a high preventive effect of LT is obtained by systemic administration in syngeneic mice, but not in nude mice. In addition, the pretreatment with anti-asialo-GM1 antibody completely prevents the antimetastatic effect of LT, but also blocks that effect of control mice without LT treatment. In conclusion, LT appears to be a potent cytokine against tumor growth and metastasis in vivo. The differences between nude and syngeneic mice suggest the involvement of host immunity in the expression of LT function.

Topics: Animals; Female; G(M1) Ganglioside; Glycosphingolipids; Glycosylation; Humans; Immunization, Passive; Lung Neoplasms; Lymphotoxin-alpha; Mammary Neoplasms, Experimental; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Nude; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms; Recombinant Proteins; Sarcoma, Experimental; Tumor Cells, Cultured

The possible role that natural killer (NK) cells might play in the control of tumors and certain infections prompted an investigation of the status of NK cells in aged mice of two inbred strains (C3H and C57BL/6). The frequency of NK cells in young-adult and aged mice was assessed by two methods that provided accurate estimates of the relative numbers of NK cells: (a) a functional assay procedure from which the number of lytic units could be estimated, and (b) a target-cell (YAC-1 tumor cells) binding procedure. The frequency of NK cells in the spleens of untreated mice as well as in mice infected with Trypanosoma musculi, the latter a powerful NK cell activating agent, was determined. In both strains of mice the frequency of functionally-competent NK cells declined significantly with age, to a greater extent in C3H than in C57BL/6 mice. Similarly, the potential to generate NK cells upon parasite activation was significantly less in aged than in young mice and the reduced potential was more apparent in C3H than in C57BL/6 mice. In contrast, the target cell-binding procedure showed only a modest decline in the frequency of NK cells in the spleens of aged mice of either strain. It appears, therefore, that the decline in NK activity during aging is a reflection of loss of competence to lyse targets rather than a major decline in the actual numbers of NK cells.

Topics: Animals; Cell Survival; G(M1) Ganglioside; Glycosphingolipids; Immune Sera; Killer Cells, Natural; Lectins; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Neoplasms; Spleen; Wheat Germ Agglutinins

The immunoreactivity of a monosialoganglioside antigen defined by monoclonal antibody 116NS19-9 (19-9) was studied in neoplastic and normal glandular and mucosal epithelia using an indirect immunoperoxidase method. In neoplastic mucosae, the antigen was detected in the majority of colorectal and endometrial carcinomas, predominantly in a focal staining pattern. A substantial proportion of gastric and pancreatic tumors and an occasional breast carcinoma also reacted with the monoclonal antibody. Expression of the monosialoganglioside in normal colonic mucosa appeared to be restricted to areas adjacent to tumor tissue. In gastric mucosa, the antigen was confined to some areas showing intestinal metaplasia. The antigen was also detected in the epithelium of normal mucosa of the gall bladder and endocervix, as well as in some ductal epithelia of the pancreas and salivary glands. Most other mucosae were negative for antigen expression.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens; Antigens, Neoplasm; Colonic Neoplasms; Female; G(M1) Ganglioside; Gangliosides; Humans; Immunoenzyme Techniques; Male; Neoplasms; Organ Specificity; Rectal Neoplasms; Uterine Neoplasms

The present study was performed to further evaluate the possible in vivo involvement of natural killer (NK) cells in host resistance against tumors. Selective depression of NK activity in Wistar Furth rats was induced by i.p. or i.v. injection of rabbit anti-asialo GM1. This antiserum has previously been shown to produce a decrease in NK activity and a parallel increase in tumor growth in mice. In the present study, rats treated with this antibody showed a parallel decrease in NK activity and in the frequency of large granular lymphocytes (LGL) in the spleen and peripheral blood, indicating that the antiserum-induced depression of NK activity in these sites was probably caused by an elimination of most effector cells. To further determine the possible role of rat LGL in tumor rejection in vivo, we studied LGL involvement in the rapid clearance of radiolabeled tumor cells from the lungs, an assay previously shown to correlate well with in vitro NK activity. Animals treated with anti-asialo GM1 antiserum were found to have a substantial decrease in the in vivo rate of clearance of tumor cells from the lungs. Furthermore, the adoptive transfer of a highly enriched population of LGL into NK-depressed animals 2 hr before tumor challenge, partially restored their cytotoxic activity against established cell lines in vitro and their ability to eliminate radiolabeled cells from the lungs. These results provide direct support for the hypothesis that NK cells are involved in in vivo resistance to tumors, particularly in the elimination of potentially metastatic tumor cells from the circulation and capillary beds.

Topics: Animals; Antibodies; Cytotoxicity, Immunologic; Female; G(M1) Ganglioside; Glycosphingolipids; Killer Cells, Natural; Lymphocytes; Male; Neoplasms; Rats

Topics: Animals; G(M1) Ganglioside; Glycosphingolipids; Humans; Immune Sera; Immunity, Innate; Killer Cells, Natural; Mice; Mice, Nude; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms; Neoplasms, Experimental; Transplantation, Heterologous