apyrase and Neoplasm-Metastasis

Some breast tumors metastasize aggressively whereas others remain dormant for years. The mechanism governing metastatic dormancy remains largely unknown. Through high-parametric single-cell mapping in mice, we identify a discrete population of CD39

Topics: Animals; Antigens, CD; Apyrase; Breast Neoplasms; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Humans; Immunotherapy; Lung; Mammary Neoplasms, Experimental; Mice, Inbred BALB C; Mice, Inbred NOD; Mice, Knockout; Neoplasm Metastasis; Programmed Cell Death 1 Receptor

UVR and immunosuppression are major risk factors for cutaneous squamous cell carcinoma (cSCC). Regulatory T cells promote cSCC carcinogenesis, and in other solid tumors, infiltrating regulatory T cells and CD8

Topics: Adenosine; Apyrase; Carcinoma, Squamous Cell; DNA Damage; DNA Repair; Forkhead Transcription Factors; Humans; Interleukin-27; Memory T Cells; Neoplasm Metastasis; Programmed Cell Death 1 Receptor; Skin Neoplasms; Ultraviolet Rays

Responses to immunotherapy vary between different cancer types and sites. Here, we aimed to investigate features of exhaustion and activation in tumor-infiltrating CD8 T cells at both the primary and metastatic sites in epithelial ovarian cancer.. Tumor tissues and peripheral blood were obtained from 65 patients with ovarian cancer. From these samples, we isolated tumor-infiltrating lymphocytes (TILs) and peripheral blood mononuclear cells. These cells were used for immunophenotype using multicolor flow cytometry, gene expression profile using RNA sequencing and ex vivo functional restoration assays.. We found that CD39. Severely exhausted PD-1

Topics: Apyrase; Carcinoma, Ovarian Epithelial; CD8-Positive T-Lymphocytes; Female; Humans; Immune Checkpoint Inhibitors; Middle Aged; Neoplasm Metastasis; Prospective Studies; Tumor Necrosis Factor Receptor Superfamily, Member 9

Purines are well-known as intracellular sources for energy but they also act as extracellular signaling molecules. In the recent years, there has been a growing interest in the therapeutic potential of purinergic signaling for cancer treatment. This is the first study to analyze lung purine levels and purinergic receptors in non-small-cell lung cancer (NSCLC) patients.. In this prospective clinical trial we enrolled 26 patients with NSCLC and 21 patients with chronic obstructive pulmonary disease (COPD) without signs of malignancy. The purine concentrations were analyzed in bronchoalveolar lavage fluid (BALF) using fluorescent/luminescent assays. Expression of purinergic receptors and ectonucleotidases were analyzed using real time quantitative polymerase chain reaction (RT-qPCR).. Patients with NSCLC have significantly lower ATP and ADP concentrations in BALF than patients with COPD (p=0.006 and p=0.009). Expression of the ectonucleotidase CD39 is significantly higher in BAL cells from cancer patients compared to COPD (p=0.001) as well as in metastasized tumors compared to non-metastasized tumors (p=0.009). Receptor-analysis revealed a higher expression of P2X4 (p=0.03), P2X7 (p=0.001) and P2Y1 (p=0.003) in BAL cells of tumors with distant metastasis.. Our data suggests a role for CD39 in lung cancer tumor microenvironment, influencing tumor invasiveness and metastasization. Potentially the increased degradation of ATP and ADP leads to a subversion of their anti-neoplastic effects. Furthermore P2Y1, P2X4 and P2X7 receptors are upregulated in BAL cells in metastatic disease. Our findings might facilitate the identification of new therapeutic targets for cancer immunotherapy.

Topics: Adenosine Diphosphate; Adenosine Triphosphate; Aged; Antigens, CD; Apyrase; Bronchoalveolar Lavage Fluid; Carcinoma, Non-Small-Cell Lung; Female; Humans; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Pulmonary Disease, Chronic Obstructive; Receptors, Purinergic; Signal Transduction; Tumor Microenvironment

CD73/ecto-5'-nucleotidase dephosphorylates extracellular AMP into adenosine, and it is a key enzyme in the regulation of adenosinergic signaling. The contribution of host CD73 to tumor growth and anti-tumor immunity has not been studied. Here, we show that under physiological conditions CD73-deficient mice had significantly elevated ATPase and ADPase activities in LN T cells. In a melanoma model, the growth of primary tumors and formation of metastasis were significantly attenuated in mice lacking CD73. Among tumor-infiltrating leukocytes there were fewer Tregs and mannose receptor-positive macrophages, and increased IFN-γ and NOS2 mRNA production in CD73-deficient mice. Treatment of tumor-bearing animals with soluble apyrase, an enzyme hydrolyzing ATP and ADP, significantly inhibited tumor growth and accumulation of intratumoral Tregs and mannose receptor-positive macrophages in the WT C57BL/6 mice but not in the CD73-deficient mice. Pharmacological inhibition of CD73 with α,β-methylene-adenosine-5'-diphosphate in WT mice retarded tumor progression similarly to the genetic deletion of CD73. Together these data show that increased pericellular ATP degradation in the absence of CD73 activity in the host cells is a novel mechanism controlling anti-tumor immunity and tumor progression, and that the purinergic balance can be manipulated therapeutically to inhibit tumor growth.

Topics: 5'-Nucleotidase; Adenosine Diphosphate; Adenosine Triphosphatases; Animals; Apyrase; Cell Proliferation; Disease Progression; Interferon-gamma; Lectins, C-Type; Lymphocytes; Lymphocytes, Tumor-Infiltrating; Macrophages; Mannose Receptor; Mannose-Binding Lectins; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Mice, Knockout; Neoplasm Metastasis; Nitric Oxide Synthase Type II; Purines; Receptors, Cell Surface; RNA, Messenger; Signal Transduction; T-Lymphocytes, Regulatory

Platelets contribute to the development of metastasis, the most common cause of mortality in cancer patients, but the precise role that anti-platelet drugs play in cancer treatment is not defined. Metastatic tumor cells can produce platelet alphaIIb beta3 activators, such as ADP and thromboxane A(2) (TXA(2)). Inhibitors of platelet beta3 integrins decrease bone metastases in mice but are associated with significant bleeding. We examined the role of a novel soluble apyrase/ADPase, APT102, and an inhibitor of TXA(2) synthesis, acetylsalicylic acid (aspirin or ASA), in mouse models of experimental bone metastases. We found that treatment with ASA and APT102 in combination (ASA + APT102), but not either drug alone, significantly decreased breast cancer and melanoma bone metastases in mice with fewer bleeding complications than observed with alphaIIb beta3 inhibition. ASA + APT102 diminished tumor cell induced platelet aggregation but did not directly alter tumor cell viability. Notably, APT102 + ASA treatment did not affect initial tumor cell distribution and similar results were observed in beta3-/- mice. These results show that treatment with ASA + APT102 decreases bone metastases without significant bleeding complications. Anti-platelet drugs such as ASA + APT102 could be valuable experimental tools for studying the role of platelet activation in metastasis as well as a therapeutic option for the prevention of bone metastases.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apyrase; Aspirin; Bone Neoplasms; Diagnostic Imaging; Melanoma, Experimental; Mice; Neoplasm Metastasis; Platelet Aggregation; Platelet Aggregation Inhibitors; Tumor Burden

Saos-2 cells, derived from a primary human osteosarcoma, caused dose-dependent platelet aggregation in heparinised human platelet-rich plasma. Saos-2 tumour cell-induced platelet aggregation (TCIPA) was completely inhibited by hirudin but unaffected by apyrase. The cell suspension shortened the plasma recalcification times of normal, factor VIII-deficient and factor IX-deficient human plasmas in a dose-dependent manner. However, the cell suspension did not affect the recalcification time of factor VII-deficient plasma. Moreover, a monoclonal antibody (MAb) against human tissue factor completely abolished TCIPA. Flow cytometric analysis using anti-integrin MAbs as the primary binding ligands demonstrated that the integrin receptors alpha v beta 3, alpha 5 beta 1 and alpha 6 beta 1 were present of Saos-2 cells, which might mediate tumour cell adhesion to extracellular matrix. Rhodostomin, an Arg-Gly-Asp (RGD)-containing snake venom peptide which antagonises the binding of fibrinogen to platelet membrane glycoprotein IIb/IIIa, prevented Saos-2 TCIPA as well as tumour cell adhesion to vitronectin, fibronectin and collagen type I. Likewise, the synthetic peptide Gly-Arg-Gly-Asp-Ser (GRGDS) showed a similar effect. On a molar basis, rhodostomin was about 18,000 and 1000 times, respectively, more potent than GRGDS in inhibiting TCIPA and tumour cell adhesion.

Topics: Amino Acid Sequence; Antibodies, Monoclonal; Apyrase; Blood Coagulation Disorders; Blood Coagulation Factors; Bone Neoplasms; Cell Adhesion; Enzyme Activation; Extracellular Matrix; Hirudins; Humans; Integrins; Molecular Sequence Data; Neoplasm Metastasis; Neoplasm Proteins; Oligopeptides; Osteosarcoma; Peptides; Platelet Aggregation; Platelet Aggregation Inhibitors; Platelet Membrane Glycoproteins; Protein Binding; Thrombin; Tumor Cells, Cultured

We established five clones (ML-01, ML-02, MH-01, MH-02, MH-03) from murine 3-methylcholanthrene-induced fibrosarcoma A (Meth A), and investigated their experimental metastatic potentials in relation to their platelet-aggregating activities. A clone with a high metastatic potential (MH-02) showed a characteristic biphasic pattern of platelet aggregation, of which the first peak was not present in the aggregation patterns of the clone with low metastatic potential (ML-01). The first peak was eliminated by treatment of the cells with apyrase, indicating that adenosine diphosphate (ADP) was the causative substance of this particular peak. The metastatic potential of clones correlated well with the ADP concentration of the culture media. These results suggest that the increased ADP production and consequential enhancement of platelet-aggregating activity are closely related to the increment of pulmonary metastatic potential of MH-02 clone.

Topics: Animals; Apyrase; Fibrosarcoma; Hot Temperature; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Microbial Collagenase; NADP; Neoplasm Metastasis; Neuraminidase; Platelet Aggregation; Trypsin; Tumor Cells, Cultured; Ultracentrifugation

Platelet aggregating activity of the NCG human neuroblastoma cell line was compared with that of the HL-60 human promyelocytic leukemia cell line. NCG, in intact cell suspensions and ultracentrifuged pellets, induced platelet aggregation most significantly in heparinized platelet rich plasma (PRP) containing 2.5 units/ml of heparin, but not in the presence of higher concentrations of heparin or 5 mM ethylenediamine-tetraacetate or in citrated PRP. NCG induced platelet aggregation was also inhibited by hirudin or (2R,4R)-4-methyl-1-[N2-(3-methyl-1,2,3,4-tetrahydro-8-quinolinesulfon yl)-L- arginyl]-2-piperidinecarboxylic acid (MD 805) in the same manner as that of tissue thromboplastin induced platelet aggregation. HL-60 cells did not induce platelet aggregation in our heparinized PRP assay systems; however, after treatment with neuraminidase HL-60 cells became active in aggregating platelets in either heparinized or citrated PRP. NCG demonstrated high procoagulant activity by either intact cell suspensions or ultracentrifuged pellets. The procoagulant activity of NCG was reduced in Factor VII deficient human plasma as it was in the results obtained by tissue thromboplastin. These results suggest that NCG induces platelet aggregation via thrombin generated through procoagulant activity which is shed in association with microvesicles demonstrated in the ultracentrifuged pellets. This type of platelet aggregating activity found in NCG is significantly different from that of HL-60.

Topics: Adenosine Diphosphate; Adenosine Triphosphate; Apyrase; Blood Coagulation; Calcium; Cell Line; Heparin; Humans; Neoplasm Metastasis; Neuraminidase; Neuroblastoma; Platelet Aggregation; Thrombin; Ultracentrifugation

Platelets may promote the development of metastasis, and tumor cells that aggregate platelets are believed to be more malignant. We studied three different human mammary carcinoma cell lines, which had different interactions with human platelet-rich plasma (PRP). The MCF-7 and the T47-D cell lines induced an adenosine diphosphate (ADP)-mediated platelet aggregation. The third cell line, MDA-MB 231 did not induce any platelet aggregation. On the contrary, this cell line inhibited ADP- and arachidonic acid-induced platelet aggregation. This inhibiting activity is mainly adenosine-mediated. The mechanism by which platelets may contribute to the dissemination of cancer could be related to platelet growth factors. MCF-7 and T47-D cell lines induced a release of platelet-derived growth factor (PDGF). On the contrary, the MDA-MB 231 cell line did not induce any platelet release. The role of these platelet growth factors in tumor cell growth is discussed.

Topics: 6-Ketoprostaglandin F1 alpha; Adenosine Diphosphate; Alprostadil; Apyrase; Arachidonic Acid; Arachidonic Acids; Blood Platelets; Breast Neoplasms; Carcinoma; Cell Line; Creatine Kinase; Humans; Neoplasm Metastasis; Platelet Aggregation; Thromboxane B2