ganglioside--gd2 and Breast-Neoplasms

Topics: AC133 Antigen; Aldehyde Dehydrogenase 1 Family; Animals; Antigens, CD; Biomarkers, Tumor; Breast Neoplasms; CD24 Antigen; CD55 Antigens; Female; Gangliosides; Glycoproteins; Hedgehog Proteins; Humans; Hyaluronan Receptors; Isoenzymes; Neoplastic Stem Cells; Octamer Transcription Factor-3; Peptides; Receptors, Notch; Retinal Dehydrogenase; Signal Transduction; Wnt Signaling Pathway

Anti-idiotype (Id) vaccine therapy has been tested and shown to be effective, in several animal models, for triggering the immune system to induce specific and protective immunity against bacterial, viral and parasitic infections. The administration of anti-Id antibodies as surrogate tumor-associated antigens (TAA) also represents another potential application of the concept of the Id network. Limited experience in human trials using anti-Id to stimulate immunity against tumors has shown promising results. In this "counter-point" article, we discuss our own findings showing the potential of anti-Id antibody vaccines to be novel therapeutic approaches to various human cancers and also discuss where anti-Id vaccines may perform better than traditional multiple-epitope antigen vaccines.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Neoplasm; Antigens, Neoplasm; Breast Neoplasms; Cancer Vaccines; Carcinoembryonic Antigen; Clinical Trials as Topic; Colorectal Neoplasms; Epitopes; Evaluation Studies as Topic; Gangliosides; Glycolipids; Glycoproteins; Humans; Immunization; Leukemia-Lymphoma, Adult T-Cell; Lipid Droplets; Melanoma; Mice; Mice, Inbred C57BL; Models, Immunological; Molecular Mimicry; Neoplasms; Neoplasms, Experimental; Treatment Outcome

Cancer stem cells (CSCs) are a small subpopulation of cancer cells that have increased resistance to conventional therapies and are capable of establishing metastasis. However, only a few biomarkers of CSCs have been identified. Here, we report that ganglioside GD2 (a glycosphingolipid) identifies a small fraction of cells in human breast cancer cell lines and patient samples that are capable of forming mammospheres and initiating tumors with as few as 10 GD2+ cells. In addition, the majority of GD2+ cells are also CD44hiCD24lo, the previously established CSC-associated cell surface phenotype. Gene expression analysis revealed that GD3 synthase (GD3S) is highly expressed in GD2+ as well as in CD44hiCD24lo cells and that interference with GD3S expression, either by shRNA or using a pharmacological inhibitor, reduced the CSC population and CSC-associated properties. GD3S knockdown completely abrogated tumor formation in vivo. Also, induction of epithelial-mesenchymal transition (EMT) in transformed human mammary epithelial cells (HMLER cells) dramatically increased GD2 as well as GD3S expression in these cells, suggesting a role of EMT in the origin of GD2+ breast CSCs. In summary, we identified GD2 as a new CSC-specific cell surface marker and GD3S as a potential therapeutic target for CSCs, with the possibility of improving survival and cure rates in patients with breast cancer.

Topics: Animals; Biomarkers, Tumor; Breast Neoplasms; CD24 Antigen; Cell Line, Transformed; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Female; Gangliosides; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Hyaluronan Receptors; Mice; Mice, Inbred NOD; Mice, SCID; Neoplasm Proteins; Neoplasm Transplantation; Neoplastic Stem Cells; Sialyltransferases; Transplantation, Heterologous

Anti-idiotype (Id) vaccine therapy has been tested and shown to be effective, in several animal models, for triggering the immune system to induce specific and protective immunity against bacterial, viral and parasitic infections. The administration of anti-Id antibodies as surrogate tumor-associated antigens (TAA) also represents another potential application of the concept of the Id network. Limited experience in human trials using anti-Id to stimulate immunity against tumors has shown promising results. In this "counter-point" article, we discuss our own findings showing the potential of anti-Id antibody vaccines to be novel therapeutic approaches to various human cancers and also discuss where anti-Id vaccines may perform better than traditional multiple-epitope antigen vaccines.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Neoplasm; Antigens, Neoplasm; Breast Neoplasms; Cancer Vaccines; Carcinoembryonic Antigen; Clinical Trials as Topic; Colorectal Neoplasms; Epitopes; Evaluation Studies as Topic; Gangliosides; Glycolipids; Glycoproteins; Humans; Immunization; Leukemia-Lymphoma, Adult T-Cell; Lipid Droplets; Melanoma; Mice; Mice, Inbred C57BL; Models, Immunological; Molecular Mimicry; Neoplasms; Neoplasms, Experimental; Treatment Outcome

Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype with no effective standard therapy. Breast cancer stem-like cells (BCSCs) in primary TNBCs are reported to be responsible for metastatic spread of the disease and resistance to chemotherapy, but no available therapeutic tools target BCSCs. We previously reported that the ganglioside GD2 is highly expressed on BCSCs and that inhibition of its expression hampers TNBC growth. We therefore hypothesized that the anti-GD2 antibody dinutuximab (ch14.18) targets GD2. To test our hypothesis, we first determined GD2 expression via immunohistochemistry in frozen primary tumor samples from patients with TNBC (n=89). Then, we examined the effects of dinutuximab on TNBC cell adhesion, migration, and mammosphere formation in vitro and on tumor growth in vivo using TNBC cell-line and patient-derived xenograft (PDX) models.. We found that GD2 was expressed in around 60% of primary TNBC tumors at variable levels and was associated with worse overall survival of patients with TNBC (p=0.002). GD2 was found to be expressed in tumors and stroma, but normal ducts and lobules in adjacent tissues have shown low or no GD2 staining, indicating that GD2 is potentially a novel biomarker for tumor and its microenvironment. Treatment with dinutuximab significantly decreased adhesion and migration of MDA-MB-231 and SUM159 TNBC cells. Moreover, dinutuximab treatment inhibited mTOR signaling, which has been shown to be regulated by GD2 in BCSCs. Dinutuximab also reduced tumor growth in nude mice bearing TNBC cell-line xenografts. Finally, dinutuximab in combination with activated natural killer cells inhibited tumor growth in a TNBC PDX model and improved overall survival of tumor-bearing mice.. Dinutuximab successfully eliminated GD2

Topics: Animals; Antibodies, Monoclonal; Antineoplastic Agents, Immunological; Breast Neoplasms; Cell Adhesion; Cell Line, Tumor; Cell Movement; Female; Gangliosides; Humans; Immunotherapy, Adoptive; Killer Cells, Natural; Mice, Nude; Mice, SCID; Neoplasm Invasiveness; Neoplastic Stem Cells; Signal Transduction; Tumor Burden; Xenograft Model Antitumor Assays

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Topics: Acetylation; Acetyltransferases; Apoptosis; Breast Neoplasms; Cell Proliferation; Female; Gangliosides; Humans; Prognosis; Survival Rate; Tumor Cells, Cultured

Topics: Breast Neoplasms; Cell Line, Tumor; Extracellular Signal-Regulated MAP Kinases; Gangliosides; Gene Expression Regulation, Neoplastic; Humans; Intercellular Adhesion Molecule-1; MCF-7 Cells; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Sialyltransferases

Breast cancer (BC) is a heterogeneous disease, including different subtypes having diverse incidence, drug-sensitivity and survival rates. In particular, claudin-low and basal-like BC have mesenchymal features with a dismal prognosis. Disialoganglioside GD2 is a typical neuroectodermal antigen expressed in a variety of cancers. Despite its potential relevance in cancer diagnostics and therapeutics, the presence and role of GD2 require further investigation, especially in BC. Therefore, we evaluated GD2 expression in a cohort of BC patients and its correlation with clinical-pathological features.Sixty-three patients with BC who underwent surgery without prior chemo- and/or radiotherapy between 2001 and 2014 were considered. Cancer specimens were analyzed by immunohistochemistry and GD2-staining was expressed according to the percentage of positive cells and by a semi-quantitative scoring system.Patient characteristics were heterogeneous by age at diagnosis, histotype, grading, tumor size, Ki-67 and receptor-status. GD2 staining revealed positive cancer cells in 59% of patients. Among them, 26 cases (41%) were labeled with score 1+ and 11 (18%) with score 2+. Notably, the majority of metaplastic carcinoma specimens stained positive for GD2. The univariate regression logistic analysis revealed a significant association of GD2 with triple-receptor negative phenotype and older age (> 78) at diagnosis.We demonstrate for the first time that GD2 is highly prevalent in a cohort of BC patients clustering on very aggressive BC subtypes, such as triple-negative and metaplastic variants.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Female; Gangliosides; Humans; Immunohistochemistry; Middle Aged; Neoplasm Grading; Neoplasm Staging; Triple Negative Breast Neoplasms

We have identified that the ganglioside GD2 is a marker for breast cancer stem cells (BCSCs), and that targeting the enzyme GD3 synthase (GD3S, which regulates GD2 biosynthesis) reduces breast tumorigenesis. The pathways regulating GD2 expression, and their anomalous functions in BCSC, are unclear. Proteomic analysis of GD2+ and GD2- cells from breast cancer cell lines revealed the activation of NFκB signaling in GD2+ cells. Dose- and time-dependent suppression of NFκB signaling by the small molecule inhibitor BMS-345541 reduced GD2+ cells by > 90%. Likewise, BMS-345541 inhibited BCSC GD3S expression, mammosphere formation, and cell migration/invasion in vitro. Breast tumor-bearing mice treated with BMS-345541 showed a statistically significant decrease in tumor volume and exhibited prolonged survival compared to control mice, with a median survival of 78 d for the BMS-345541-treated group vs. 58 d for the controls. Moreover, in an experimental metastases model, treatment with BMS-345541 reduced the lung metastases by > 5-fold. These data suggest that GD2 expression and function,and NFκB signaling, are related, and they control BCSCs tumorigenic characteristics. Thus, the suppression of NFκB signaling by BMS-345541 is a potentially important advance in controlling breast cancer growth and metastases.

Topics: Animals; Breast Neoplasms; Carcinogenesis; Cell Line, Tumor; Gangliosides; Gene Expression Regulation, Neoplastic; Humans; I-kappa B Kinase; Imidazoles; Mice, Inbred NOD; Mice, Knockout; Mice, SCID; Neoplasm Metastasis; Neoplastic Stem Cells; Quinoxalines; RNA Interference; Sialyltransferases; Signal Transduction; Xenograft Model Antitumor Assays

Two major obstacles in developing cancer vaccines are identifying unique tumor-associated antigens (TAA) and overcoming the lack of structural information about TAAs. Unlike progress with T cell-based vaccines, B cell vaccines are less well developed due to discontinuous or spatially disposed B cell epitopes. Synthetic peptides that emulate B cell epitopes are nevertheless proposed to induce immune responses to TAA. Currently, such antigen-mimicking peptides are identified using informatics approaches, by screening of random peptide libraries against an isolating antibody without any regard to structural principles and by rationale design methodologies. In our own studies we have developed various peptide mimics of TAAs based on combining the structural analysis of antibody-antigen complexes with the peptide library screening process. Understanding the structural context of antigen mimicry is key, as our studies show that mimicry depends on the structural and conformational features of the combining region of antibody-antigen surface amino acids. The ability of a mimic to contact the same set of amino acids found on a template antibody dictates whether or not it is a functional antigenic mimic capable of inducing TAA cross-reactive antibodies. Presented here is an overview of our current rationale and status of structure-based tumor antigenic mimics relevant for breast cancer TAAs.

Topics: Antigens, Neoplasm; Breast Neoplasms; Cancer Vaccines; Cell Surface Display Techniques; Epitopes, B-Lymphocyte; Female; Gangliosides; Humans; Lewis Blood Group Antigens; Molecular Mimicry; Receptor, ErbB-2

We have recently established and characterized cellular clones deriving from MDA-MB-231 breast cancer cells that express the human G(D3) synthase (GD3S), the enzyme that controls the biosynthesis of b- and c-series gangliosides. The GD3S positive clones show a proliferative phenotype in the absence of serum or growth factors and an increased tumor growth in severe immunodeficient mice. This phenotype results from the constitutive activation of the receptor tyrosine kinase c-Met in spite of the absence of ligand and subsequent activation of mitogen-activated protein kinase/extracellular signal-regulated kinase and phosphoinositide 3-kinase/Akt pathways. Here, we show by mass spectrometry analysis of total glycosphingolipids that G(D3) and G(D2) are the main gangliosides expressed by the GD3S positive clones. Moreover, G(D2) colocalized with c-Met at the plasma membrane and small interfering RNA silencing of the G(M2)/G(D2) synthase efficiently reduced the expression of G(D2) as well as c-Met phosphorylation and reversed the proliferative phenotype. Competition assays using anti-G(D2) monoclonal antibodies also inhibit proliferation and c-Met phosphorylation of GD3S positive clones in serum-free conditions. Altogether, these results demonstrate the involvement of the disialoganglioside G(D2) in MDA-MB-231 cell proliferation via the constitutive activation of c-Met. The accumulation of G(D2) in c-Met expressing cells could therefore reinforce the tumorigenicity and aggressiveness of breast cancer tumors.

Topics: Antibodies, Monoclonal; Breast Neoplasms; Cell Membrane; Cell Proliferation; Gangliosides; Humans; Mass Spectrometry; Phenotype; Phosphorylation; Proto-Oncogene Proteins c-met; RNA Interference; RNA, Small Interfering; Sialyltransferases; Tumor Cells, Cultured

The bone marrow microenvironment is considered a critical component in the dissemination and fate of cancer cells in the metastatic process. We explored the possible correlation between bone marrow mesenchymal stem cells (BM-MSC) and disseminated breast cancer-initiating cells (BCIC) in primary breast cancer patients.. Bone marrow mononuclear cells (BM-MNC) were collected at the time of primary surgery in 12 breast cancer patients. BM-MNC was immunophenotyped and BCIC was defined as epithelial cells (CD326+CD45-) with a "stem-like" phenotype (CD44+CD24low/-, ALDH activity). BM-MSC was defined as CD34-CD45-cells that co-expressed GD2, CD271, and/or CD200 within CD326-depleted BM-MNC.. The percentages of BCIC (Aldefluor+CD326+CD44+CD24-) correlated with the percentages of BM-MSC, either CD45-GD2+CD200+CD271+ (Kedall's tau = 0.684, p = 0.004) or CD45-GD2+CD271+ in the bone marrow (Kedall's tau = 0.464, p = 0.042).. There was a positive correlation between mesenchymal stem cells expressing GD2 and CD271 and breast cancer-initiating cells in BM of patients with primary breast cancer.

Topics: Adult; Aged; Antigens, CD; Bone Marrow Cells; Breast Neoplasms; Female; Gangliosides; Humans; Immunophenotyping; Mesenchymal Stem Cells; Middle Aged