apyrase and Melanoma

The mechanisms leading to immune escape of melanoma have been largely investigated in relation to its tumour immunogenicity and features of inflamed microenvironment that promote the immune suppression during the disease progression. These findings have recently led to advantages in terms of immunotherapy-based approaches as rationale for overcoming the immune escape. However, besides immune checkpoints, other mechanisms including the adenosine produced by ectonucleotidases CD39 and CD73 contribute to the melanoma progression due to the immunosuppression induced by the tumour milieu. On the other hand, CD73 has recently emerged as both promising therapeutic target and unfavourable prognostic biomarker. Here, we review the major mechanisms of immune escape activated by the CD39/CD73/adenosine pathway in melanoma and focus potential therapeutic strategies based on the control of CD39/CD73 downstream adenosine receptor signalling. These evidences provide the basis for translational strategies of immune combination, while CD73 would serve as potential prognostic biomarker in metastatic melanoma.

Topics: 5'-Nucleotidase; Adenosine; Apyrase; Biomarkers, Tumor; GPI-Linked Proteins; Humans; Immune Tolerance; Immunosuppression Therapy; Immunotherapy; Melanoma; Receptors, Purinergic P1; Signal Transduction; Tumor Microenvironment

Melanoma cancer cell proliferation, motility, invasion, and tumor growth is affected by the adenosine pathway that consists of adenosine-synthesizing enzymes, receptors, and their respective agonists/antagonists. Accumulating evidence suggests that ischemia and inflammation, two conditions associated with melanoma, display dysregulated adenosine metabolism, which implicates it as the mechanism responsible for the pathogenesis of melanoma, thereby resulting in advanced diagnosis and therapy. Suppression of adenosine signaling by inhibiting adenosine receptors or adenosine-generating enzymes (CD39 and CD73) on melanoma cells presents a novel therapeutic target for patients with melanoma. This review summarizes the role of adenosine signaling in the pathogenesis of melanoma to advance its understanding and hence improve therapeutics and management.

Topics: 5'-Nucleotidase; Adenosine; Apyrase; GPI-Linked Proteins; Humans; Melanoma; Signal Transduction

Extracellular adenosine is produced from ATP by CD39 and CD73, and can modulate tumor development by acting on cancer cells or immune cells. Adenosine metabolism has been poorly studied in uveal melanoma. We studied the protein levels of CD39 and CD73 in a small, well described cohort of patients with uveal melanoma. Our results show a high variability in the levels of the two proteins, both in positivity and in intensity. Our results suggest that similar studies on larger cohorts could determine the clinical value and the druggability of these enzymes in the given clinical setting.Supplemental data for this article is available online at http://dx.doi.org/10.1080/15257770.2022.2032738.

Topics: 5'-Nucleotidase; Adenosine; Adenosine Triphosphate; Antigens, CD; Apyrase; Humans; Melanoma

Adjuvant immune checkpoint inhibitor (ICI) immunotherapies have significantly reduced the recurrence rate in high-risk patients with stage III melanoma compared with surgery alone. However, 48% of anti-PD-1-treated patients will develop recurrent disease within 4 years. There is a need to identify biomarkers of recurrence after adjuvant ICI to enable identification of patients in need of alternative treatment strategies. As cytotoxic T cells are critical for the antitumor response to anti-PD-1, we sought to determine whether specific subsets were predictive of recurrence in anti-PD-1-treated high-risk patients with stage III melanoma.. Associations with recurrence in patients with stage III melanoma were sought by analyzing resection specimens (n=103) taken prior to adjuvant nivolumab/pembrolizumab±low-dose/low-interval ipilimumab. Multiplex immunohistochemistry was used to quantify intratumoral CD8+ T-cell populations using phenotypical markers CD39, CD103, and PD-1.. With a median follow-up of 19.3 months, 37/103 (36%) of patients had a recurrence. Two CD8+ T-cell subpopulations were significantly associated with recurrence. First, CD39+ tumor-resident memory cells (CD39+CD103+PD-1+CD8+ (CD39+ Trm)) comprised a significantly higher proportion of CD8+ T cells in recurrence-free patients (p=0.0004). Conversely, bystander T cells (CD39-CD103-PD-1-CD8+) comprised a significantly greater proportion of T cells in patients who developed recurrence (p=0.0002). Spatial analysis identified that CD39+ Trms localized significantly closer to melanoma cells than bystander T cells. Multivariable analysis confirmed significantly improved recurrence-free survival (RFS) in patients with a high proportion of intratumoral CD39+ Trms (1-year RFS high 78.1% vs low 49.9%, HR 0.32, 95% CI 0.15 to 0.69), no complete lymph node dissection performed, and less advanced disease stage (HR 2.85, 95% CI 1.13 to 7.19, and HR 1.29, 95% CI 0.59 to 2.82). The final Cox regression model identified patients who developed recurrence with an area under the curve of 75.9% in the discovery cohort and 69.5% in a separate validation cohort (n=33) to predict recurrence status at 1 year.. Adjuvant immunotherapy-treated patients with a high proportion of CD39+ Trms in their baseline melanoma resection have a significantly reduced risk of melanoma recurrence. This population of T cells may not only represent a biomarker of RFS following anti-PD-1 therapy, but may also be an avenue for therapeutic manipulation and enhancing outcomes for immunotherapy-treated patients with cancer.

Topics: Apyrase; Humans; Immunotherapy; Melanoma; Melanoma, Cutaneous Malignant; Programmed Cell Death 1 Receptor; Skin Neoplasms

In the context of adoptive T cell transfer (ACT) for cancer treatment, it is crucial to generate

Topics: Adoptive Transfer; Animals; Antigens, CD; Apyrase; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cell Separation; Female; Humans; Lymphocytes, Tumor-Infiltrating; Melanoma; Mice; Mice, Inbred C57BL; Phenotype; Programmed Cell Death 1 Receptor; Receptors, CXCR5

Adoptive T cell therapy (ACT) using ex vivo-expanded autologous tumor-infiltrating lymphocytes (TILs) can mediate complete regression of certain human cancers. The impact of TIL phenotypes on clinical success of TIL-ACT is currently unclear. Using high-dimensional analysis of human ACT products, we identified a memory-progenitor CD39-negative stem-like phenotype (CD39

Topics: Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Apyrase; CD8-Positive T-Lymphocytes; Female; Humans; Immunotherapy, Adoptive; Lectins, C-Type; Lymphocytes, Tumor-Infiltrating; Melanoma; Mice; Mice, Mutant Strains; Skin Neoplasms

Immune checkpoint inhibitors have revolutionized cancer treatment. However, many cancers are resistant to ICIs, and the targeting of additional inhibitory signals is crucial for limiting tumor evasion. The production of adenosine via the sequential activity of CD39 and CD73 ectoenzymes participates to the generation of an immunosuppressive tumor microenvironment. In order to disrupt the adenosine pathway, we generated two antibodies, IPH5201 and IPH5301, targeting human membrane-associated and soluble forms of CD39 and CD73, respectively, and efficiently blocking the hydrolysis of immunogenic ATP into immunosuppressive adenosine. These antibodies promoted antitumor immunity by stimulating dendritic cells and macrophages and by restoring the activation of T cells isolated from cancer patients. In a human CD39 knockin mouse preclinical model, IPH5201 increased the anti-tumor activity of the ATP-inducing chemotherapeutic drug oxaliplatin. These results support the use of anti-CD39 and anti-CD73 monoclonal antibodies and their combination with immune checkpoint inhibitors and chemotherapies in cancer.

Topics: 5'-Nucleotidase; Adenosine; Adenosine Triphosphate; Animals; Antibodies, Blocking; Antigens, CD; Antineoplastic Agents; Apyrase; Cell Line, Tumor; Disease Models, Animal; Gene Knock-In Techniques; Humans; Leukocytes, Mononuclear; Melanoma; Mice; Mice, Inbred C57BL; Mice, Knockout; Oxaliplatin; Survival Rate; T-Lymphocytes; Tumor Microenvironment

The ability of CD8

Topics: Adenosine Triphosphate; Animals; Antigens, CD; Apyrase; Breast Neoplasms; CD8-Positive T-Lymphocytes; Cytokines; Female; Humans; Lymphatic Metastasis; Lymphocytes, Tumor-Infiltrating; Male; Melanoma; Mice, Inbred BALB C; Mice, Knockout; Tumor Microenvironment; Xenograft Model Antitumor Assays

Identifying tumor antigen-specific T cells from cancer patients has important implications for immunotherapy diagnostics and therapeutics. Here, we show that CD103

Topics: Adenocarcinoma of Lung; Antigens, CD; Apyrase; Carcinoma, Squamous Cell; CD8 Antigens; CD8-Positive T-Lymphocytes; Female; Humans; Immunophenotyping; Integrin alpha Chains; Lymphocytes, Tumor-Infiltrating; Male; Melanoma; Ovarian Neoplasms; Receptors, Antigen, T-Cell, alpha-beta; Squamous Cell Carcinoma of Head and Neck; Survival Analysis; Transcriptome

Treatment of cancer has been revolutionized by immune checkpoint blockade therapies. Despite the high rate of response in advanced melanoma, the majority of patients succumb to disease. To identify factors associated with success or failure of checkpoint therapy, we profiled transcriptomes of 16,291 individual immune cells from 48 tumor samples of melanoma patients treated with checkpoint inhibitors. Two distinct states of CD8

Topics: Animals; Antibodies, Monoclonal, Humanized; Antigens, CD; Antineoplastic Agents, Immunological; Apyrase; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Humans; Immunotherapy; Leukocyte Common Antigens; Melanoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; T Cell Transcription Factor 1; Transcriptome

We have developed an array-based resequencing method to determine genetic alterations in putative cancer genes. The method relies on that the specificity of DNA polymerase in allele-specific extensions can be enhanced by terminating the extension reactions with apyrase and that a tiling set of primers are synthesized covering the investigated gene sequence. We report on such apyrase-mediated allele-specific primer extension (AMASE) assay as a method suitable for high-throughout resequencing and mutation detection in tumor suppressor genes and oncogenes. In the experimental setup, primers complementary to codons 12, 13 and codon 61 of the N-ras proto-oncogene were spotted onto glass slides. Overlapping sense and anti-sense primers were designed so that complementary primers for all possible mutations in each base position were investigated. The extension reactions were performed in a single step following hybridization of target DNA to the immobilized primers on the array surface. Mutation detection limits and the possibility of quantifying the mutations were investigated using synthetic oligonucleotides. In addition, 64 clinical samples were sequenced and 16 of these showed mutations in the N-ras gene.

Topics: Alleles; Apyrase; DNA Mutational Analysis; DNA Primers; Genes, ras; Humans; Melanoma; Oligonucleotide Array Sequence Analysis; Polymerase Chain Reaction; Proto-Oncogene Mas; Proto-Oncogenes; Sequence Analysis, DNA

Ecto-ATPase activities of melanocytes and human melanoma cell lines differing in the stage of progression were compared. A dramatic increase in ecto-ATPase activity above the level of normal melanocytes was demonstrated in the differentiated melanomas and was followed by a gradual decrease with tumor progression. The characteristics of this enzymatic activity were consistent with CD39/ecto-ATP diphosphohydrolase (ATPDase) which was found to be the major ecto-ATP-hydrolysing enzyme in melanomas. Indeed, the expression of CD39 and the level of CD39 mRNA followed a similar pattern. Since CD39 is known to regulate homotypic adhesion and, supposedly, affects the disaggregation step, we suggest that overexpression of CD39 may enable tumor cells to reduce contacts with T-lymphocytes and escape from immunological recognition.

Topics: Adenosine Triphosphatases; Antigens, CD; Apyrase; Biomarkers, Tumor; Cell Differentiation; Cell Line; Disease Progression; Enzyme Inhibitors; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Melanocytes; Melanoma; RNA, Messenger; RNA, Neoplasm; Tumor Cells, Cultured

We studied the effects on platelet function of cells isolated from freshly dissociated human tumor tissues (11 breast carcinomas, 9 colon carcinomas and 1 lymph node metastasis from melanoma) obtained at surgery as compared with cultured human tumor cells: namely, human melanoma 1402 cell line derived from a primary tumor and two lines derived from lymph node metastases (ME 7110/2 and Me 665/1) as well as a human hepatoma cell line (Hep G2). The three melanoma cell lines activated platelets by producing ADP, as evidenced by the inhibitory effect of apyrase and by the direct measurement of the agonist in the supernatants of tumor cell suspensions; this production was much greater by the cells derived from metastases than by the cells derived from the primary tumor. On the other hand, aggregation induced by Hep G2 hepatoma cells was unaffected by apyrase and was inhibited by hirudin or concanavalin A, suggesting that the cells aggregate platelets by producing thrombin, probably through tissue factor activity of the cells themselves. Cells isolated from 16 of the 21 human tumor tissues possessed a potent platelet-aggregating effect, which was not inhibited by apyrase, hirudin or concanavalin A, but was virtually abolished by the cysteine protease inhibitors iodoacetic acid or p-hydroxymercuri-phenylsulfonate. Collectively, our data demonstrate that cells isolated from freshly dissociated tumor tissues activate platelets through tumor-associated cysteine proteinases rather than by the ADP- or thrombin-dependent mechanisms characteristic of cultured human tumor cell lines.

Topics: Adenosine Diphosphate; Adult; Apyrase; Blood Platelets; Breast Neoplasms; Cell Communication; Cell Separation; Colonic Neoplasms; Concanavalin A; Cysteine Endopeptidases; Cysteine Proteinase Inhibitors; Hirudins; Humans; Melanoma; Platelet Aggregation; Tumor Cells, Cultured

Topics: Adenocarcinoma; Animals; Apyrase; Cell Line; Colonic Neoplasms; Glioma; Hirudins; Humans; Kinetics; Lung Neoplasms; Melanoma; Mesothelioma; Mice; Neoplasms; Neoplasms, Experimental; Neuroblastoma; Phospholipases; Phosphoric Monoester Hydrolases; Platelet Aggregation