heparitin-sulfate and Prostatic-Neoplasms

Functional Tregs play a key role in tumor development and progression, representing a major barrier to anticancer immunity. The mechanisms by which Tregs are generated in cancer and the influence of the tumor microenvironment on these processes remain incompletely understood. Herein, by using NMR, chemoenzymatic structural assays and a plethora of in vitro and in vivo functional analyses, we demonstrate that the tumoral carbohydrate A10 (Ca10), a cell-surface carbohydrate derived from Ehrlich's tumor (ET) cells, is a heparan sulfate-related proteoglycan that enhances glycolysis and promotes the development of tolerogenic features in human DCs. Ca10-stimulated human DCs generate highly suppressive Tregs by mechanisms partially dependent on metabolic reprogramming, PD-L1, IL-10, and IDO. Ca10 also reprograms the differentiation of human monocytes into DCs with tolerogenic features. In solid ET-bearing mice, we found positive correlations between Ca10 serum levels, tumor size and splenic Treg numbers. Administration of isolated Ca10 also increases the proportion of splenic Tregs in tumor-free mice. Remarkably, we provide evidence supporting the presence of a circulating human Ca10 counterpart (Ca10H) and show, for the first time, that serum levels of Ca10H are increased in patients suffering from different cancer types compared to healthy individuals. Of note, these levels are higher in prostate cancer patients with bone metastases than in prostate cancer patients without metastases. Collectively, we reveal novel molecular mechanisms by which heparan sulfate-related structures associated with tumor cells promote the generation of functional Tregs in cancer. The discovery of this novel structural-functional relationship may open new avenues of research with important clinical implications in cancer treatment.

Topics: Animals; Dendritic Cells; Glycosaminoglycans; Heparitin Sulfate; Humans; Male; Mice; Prostatic Neoplasms; T-Lymphocytes, Regulatory; Tumor Microenvironment

Prostate stem/progenitor cells (PrSCs) are responsible for adult prostate tissue homeostasis and regeneration. However, the related regulatory mechanisms are not completely understood. In this study, we examined the role of heparan sulfate (HS) in PrSC self-renewal and prostate regeneration. Using an in vitro prostate sphere formation assay, we found that deletion of the glycosyltransferase exostosin 1 (Ext1) abolished HS expression in PrSCs and disrupted their ability to self-renew. In associated studies, we observed that HS loss inhibited p63 and CK5 expression, reduced the number of p63+- or CK5+-expressing stem/progenitor cells, elevated CK8+ expression and the number of differentiated CK8+ luminal cells and arrested the spheroid cells in the G1/G0 phase of cell cycle. Mechanistically, HS expressed by PrSCs (in cis) or by neighboring cells (in trans) could maintain sphere formation. Furthermore, HS deficiency upregulated transforming growth factor β (TGFβ) signaling and inhibiting TGFβ signaling partially restored the sphere-formation activity of the HS-deficient PrSCs. In an in vivo prostate regeneration assay, simultaneous loss of HS in both epithelial cell and stromal cell compartments attenuated prostate tissue regeneration, whereas the retention of HS expression in either of the two cellular compartments was sufficient to sustain prostate tissue regeneration. We conclude that HS preserves self-renewal of adult PrSCs by inhibiting TGFβ signaling and functions both in cis and in trans to maintain prostate homeostasis and to support prostate regeneration.

Topics: Animals; Heparitin Sulfate; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Prostatic Neoplasms; Signal Transduction; Stem Cells; Transforming Growth Factor beta

Prostate cancer is impacting many men globally. It is a disease that has no effective treatment is available in the market. The understanding of the biophysical and biochemical aspects of the disease and the mechanism that allow it to metastasize is key to finding an effect treatment. Maintenance or pretreatment drug as well as a post treatment drug can be effective to avoid or delay the disease from appearing. The polysaccharides and monosaccharides polymers combined with vitamins can be the ingredient to developing the treatment. There are many evidences that investigators examined the individual components of the therapy proposed but never a combination of all these therapies. The one item that is not discussed is how to formulate the ingredient into an effective form which is a proprietary work being conducted currently. Nevertheless, the hypothesis seems reasonable to us and worth sharing with the scientific community.

Topics: Adenocarcinoma; Amygdalin; Antineoplastic Combined Chemotherapy Protocols; Cell Division; Drug Combinations; Drug Synergism; Extracellular Matrix; Glycosaminoglycans; Heparitin Sulfate; Humans; Injections, Intravenous; Male; Models, Biological; Myocytes, Smooth Muscle; Polysaccharides; Prostate; Prostatic Neoplasms; Vitamin A; Vitamin E

Activation of myofibroblast rich stroma is a rate-limiting step essential for cancer progression. The responsible factors are not fully understood, but TGFβ1 is probably critical. A proportion of TGFβ1 is associated with extracellular nano-vesicles termed exosomes, secreted by carcinoma cells, and the relative importance of soluble and vesicular TGFβ in stromal activation is presented. Prostate cancer exosomes triggered TGFβ1-dependent fibroblast differentiation, to a distinctive myofibroblast phenotype resembling stromal cells isolated from cancerous prostate tissue; supporting angiogenesis in vitro and accelerating tumour growth in vivo. Myofibroblasts generated using soluble TGFβ1 were not pro-angiogenic or tumour-promoting. Cleaving heparan sulphate side chains from the exosome surface had no impact on TGFβ levels yet attenuated SMAD-dependent signalling and myofibroblastic differentiation. Eliminating exosomes from the cancer cell secretome, targeting Rab27a, abolished differentiation and lead to failure in stroma-assisted tumour growth in vivo. Exosomal TGFβ1 is therefore required for the formation of tumour-promoting stroma.

Topics: Animals; Cell Differentiation; Cell Line, Tumor; Cells, Cultured; Exosomes; Gene Knockdown Techniques; Heparitin Sulfate; Human Umbilical Vein Endothelial Cells; Humans; Immunoblotting; Intercellular Signaling Peptides and Proteins; Male; Mice, Nude; Myofibroblasts; Prostatic Neoplasms; rab GTP-Binding Proteins; rab27 GTP-Binding Proteins; Stromal Cells; Transforming Growth Factor beta1; Transplantation, Heterologous

NKp44 is a natural cytotoxicity receptor expressed by human NK cells upon activation. In this study, we demonstrate that cell surface heparan sulfate proteoglycans (HSPGs), expressed by target cells, are involved in the recognition of tumor cells by NKp44. NKp44 showed heparan sulfate-dependent binding to tumor cells; this binding was partially blocked with an antibody to heparan sulfate. In addition, direct binding of NKp44 to heparin was observed, and soluble heparin/heparan sulfate enhanced the secretion of IFNgamma by NK92 cells activated with anti-NKp44 monoclonal antibody. Basic amino acids, predicted to constitute the putative heparin/heparan sulfate binding site of NKp44, were mutated. Tumor cell recognition of the mutated NKp44 proteins was significantly reduced and correlated with their lower recognition of heparin. We previously reported that NKp44 recognizes the hemagglutinin of influenza virus (IV). Nevertheless, the ability of the mutated NKp44 proteins to bind viral hemagglutinin expressed by IV-infected cells was not affected. Thus, we suggest that heparan sulfate epitope(s) are ligands/co-ligands of NKp44 and are involved in its tumor recognition ability.

Topics: Amino Acid Sequence; Animals; Antibodies, Monoclonal; Binding Sites; Carcinoma, Ductal; Cell Line, Tumor; CHO Cells; Cricetinae; Cricetulus; Epitopes; HeLa Cells; Heparitin Sulfate; Humans; Immunoglobulins; Interferon-gamma; Killer Cells, Natural; Ligands; Male; Melanoma; Molecular Sequence Data; Mutation; Natural Cytotoxicity Triggering Receptor 2; Pancreatic Neoplasms; Prostatic Neoplasms; Protein Binding; Protein Structure, Tertiary; Receptors, Immunologic; Recombinant Fusion Proteins

To investigate changes in glycosaminoglycan (GAG) profiles in human prostatic tissue.. Seventeen human prostatic samples were examined; five were normal, six hyperplastic and six were cancerous. Prostatic proteoglycans were extracted with 4 mol/L guanidine-HCl containing protease inhibitors. After digestion of the freeze-dried proteoglycan extract with papain, the prostatic GAGs were purified. Total GAGs were measured by a modified dimethylmethylene blue (DMB) method. High-performance liquid chromatography (HPLC) was used to quantify the extracted GAGs.. Six types of GAGs, i.e. chondroitin 4-sulphate (Ch-4S), chondroitin 6-sulphate (Ch-6S), dermatan sulphate (DS), chondroitin, heparan sulphate and hyaluronic acid, were analysed qualitatively and quantitatively. The total amount of GAGs was increased in hyperplastic and cancerous prostates, with the predominant components being DS and Ch-6S in these specimens. The Ch-S:DS ratio in cancerous prostate was significantly higher than that in normal and BPH tissue (P < 0.05). Moreover, chondroitin was increased in hyperplastic prostatic tissue (P < 0.01).. These results suggest that an increase in chondroitin levels may be associated with hyperplastic change and an increase in the Ch-S:DS ratio may be related to the development of malignancy.

Topics: Adult; Aged; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Dermatan Sulfate; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Male; Middle Aged; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms

There is increasing evidence that the association of prostate carcinoma cells with the surrounding extracellular matrix is important for their growth. Proteoglycans (PGs) are components of matrix that form important associations with other molecules. Heparan sulfate proteoglycan (HSPG), for example, associates with several matrix components such as fibronectin, laminin, and basic fibroblast growth factor. The purpose of this study was to determine if human prostate carcinoma cells (PC-3) synthesized HSPG in tissue culture. Three areas of sulfate-labeled matrix were collected from PC-3 cells and analyzed: attachment sites, nonattachment cell surfaces, and media. Charged groups were separated by ion-exchange columns, and PGs identified by gel filtration chromatography after chemical and enzymatic treatment. The media fraction contained the greatest proportion of sulfated PGs (79.3%), of which 45.5% were HSPG. The greatest concentration of HSPG was in the attachment site fraction where 88.6% of PGs were HSPG, although only 11.7% of sulfated molecules were PGs. In nonattachment surfaces, only 19.6% were PGs, of which 19.4% were HSPG. When PGs were assessed for hydrophobic binding to octyl-Sepharose beads, only a proportion of HSPG in the media fraction contained hydrophobic domains (18.8%). In summary, PC-3 cells synthesize at least two types of HSPG, sorting them into different matrix compartments. The major PG in attachment sites is HSPG without a hydrophobic domain, while an HSPG in the media fraction has a hydrophobic domain. The localization of different types of HSPG may be functionally important and may be altered during the metastatic process for prostate carcinoma when in association with specific stroma.

Topics: Adenocarcinoma; Chromatography, Ion Exchange; Extracellular Matrix; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Male; Prostatic Neoplasms; Proteoglycans; Tumor Cells, Cultured

The status of the basement membrane in prostatic intraepithelial neoplasia (PIN) and adenocarcinoma is unsettled. Previous studies using antibodies directed against Type IV collagen have been hindered by intense staining around the stromal smooth muscle fibers, making interpretation of acinar staining difficult. We employed a monoclonal antibody to heparan sulfate proteoglycan (HSPG) to overcome this problem, recognizing that HSPG is present in the basement membrane of epithelial and endothelial cells, but not stromal smooth muscle cells. In 22 totally-embedded whole mount radical prostatectomies for adenocarcinoma which contained PIN, intense HSPG immunoreactivity was observed in the basement membrane of all normal and hyperplastic acini, 98% of acini with high grade PIN, and 100% of acini of well differentiated (Gleason score 5) adenocarcinoma; vessels served as the internal positive control, with consistent staining throughout each specimen. The extent of HSPG immunoreactivity in cancer decreased with increasing Gleason grade (measured as percent of acini staining, in 10% increments; p = 0.002). These findings indicate that HSPG is a consistent component of the basement membrane of benign, hyperplastic, and early neoplastic prostatic acini, and, unlike other extracellular matrix proteins such as type IV collagen, is not hindered by background staining around stromal smooth muscle cells. High grade PIN and well differentiated adenocarcinoma usually maintain an intact basement membrane, and loss of the basement membrane occurs with histologic dedifferentiation.

Topics: Adenocarcinoma; Aged; Antibodies, Monoclonal; Basement Membrane; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Male; Middle Aged; Neoplasm Staging; Neoplasms, Multiple Primary; Prostatic Intraepithelial Neoplasia; Prostatic Neoplasms; Proteoglycans

This unit has in the past evaluated the Nb rat prostatic adenocarcinoma model with respect to chemotherapies. Recently, this unit has been evaluating agents that may have a role in decreasing metastatic rate. The three androgen-insensitive tumors, Nb Pr A.I. I, II, III, have been evaluated herein. Agents that have been used include indomethacin, heparin, heparin plus cortisone, and heparan sulfate (SP54). It has been shown that these agents do play a role in reducing the metastasis. In evaluation of Nb Pr A.I, I, control animals had a metastatic rate of 57%. In treatment with indocin, only 21% of the animals, three of 14 animals treated, had a metastasis, and treatment with heparin and cortisone resulted in one of 14 animals having metastasis. Similar observations were seen when treatment with SP54 and heparin was evaluated in the Nb Pr A.I. III; 18 and 27% of treatment group animals had metastasis, whereas 55% of control groups had metastasis. Similarly, in the Nb Pr A.I. II evaluation, control animals having a metastatic rate of 43% had heparin plus cortisone and heparin alone, and this particular tumor model revealed complete resolution with no animals having metastatic disease. The majority of these agents have not effected tumor volume in terms of reduction as much as the best chemotherapeutic agents in this model system include cyclophosphamide and cis-platinum.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Cortisone; Disease Models, Animal; Drug Therapy, Combination; Heparin; Heparitin Sulfate; Indomethacin; Male; Neoplasm Metastasis; Prostatic Neoplasms; Rats; Rats, Inbred Strains

The glycosaminoglycans of normal, benign hyperplastic and cancerous prostate were studied. In both prostatic hyperplasia and cancer the chondroitin sulfate:dermatan sulfate ratio was increased. In prostatic cancer this increase correlated with both the differentiation and extent of cancer in the prostate. The percentages heparan sulfate and heparan sulfate sulfation were decreased in prostatic cancer. Hyaluronic acid increased with dedifferentiation of the cancer. Histochemically, sulfated glycosaminoglycans were concentrated in the prostatic stroma at the stromal-epithelial interface. The increased chondroitin sulfate:dermatan sulfate ratio may be a nonspecific response or requirement for epithelial growth.

Topics: Adult; Aged; Chondroitin Sulfates; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Histocytochemistry; Humans; Hyaluronic Acid; Male; Middle Aged; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Uronic Acids