g(m2)-ganglioside and Skin-Neoplasms

Topics: Adjuvants, Immunologic; Antineoplastic Agents; Cancer Vaccines; Chemotherapy, Adjuvant; Clinical Trials as Topic; Disease-Free Survival; Dose-Response Relationship, Immunologic; Drug Combinations; Female; G(M2) Ganglioside; Humans; Interferon Type I; Interferon-gamma; Interleukin-2; Male; Melanoma; Multicenter Studies as Topic; Neoplasm Staging; Recombinant Proteins; Skin Neoplasms; Vaccines, Conjugate

We evaluated Eastern Cooperative Group phase II and III trials E2696 and E1694 to assess the incidence and prognostic significance of autoimmunity induced by adjuvant high-dose interferon-α2b (HDI). In E2696, patients with resectable high-risk melanoma were randomized to receive vaccination with GM2-KLH/QS-1 (GMK) plus concurrent HDI, GMK plus sequential HDI, or GMK alone. E1694 randomized patients to either HDI or GMK. Sera from 103 patients in E2696 and 691 patients in E1694 banked at baseline and up to three subsequent time points were tested by ELISA for the development of five autoantibodies. In E2696, autoantibodies were induced in 16 patients (23.2%; n=69) receiving HDI and GMK and two patients (5.9%; n=34) receiving GMK alone (P=0.031). Of 691 patients in E1694, 67 (19.1%) who received HDI (n=350) developed autoantibodies, but only 16 patients (4.7%) developed autoantibodies in the vaccine group (n=341; P<0.001). Almost all induced autoantibodies were detected at ≥12 weeks after the initiation of therapy. A 1-year landmark analysis among resected stage III patients treated with HDI in E1694 showed a trend toward a survival advantage associated with HDI-induced autoimmunity (hazard ratio=0.80; 95% confidence interval: 0.50-1.98; P=0.33). Therefore, adjuvant HDI therapy is associated with the induction of autoimmunity that should be further investigated prospectively as a surrogate marker of adjuvant therapeutic benefit. This potential biomarker develops over the course of up to 1 year, and cannot be used to alter the course of therapy. Studies of the genetic determinants of this response may better discriminate patients more likely to benefit from HDI immunomodulatory therapy.

Topics: Autoantibodies; Autoimmunity; Cancer Vaccines; Chemotherapy, Adjuvant; Disease-Free Survival; Enzyme-Linked Immunosorbent Assay; G(M2) Ganglioside; Hemocyanins; Humans; Interferon-alpha; Melanoma; Prognosis; Skin Neoplasms

The GM2 ganglioside is an antigen expressed in the majority of melanomas. The GM2-KLH/QS-21 vaccine induces high immunoglobulin M (IgM) and IgG antibody responses. The EORTC 18961 trial compared the efficacy of GM2-KLH/QS-21 vaccination versus observation.. A total of 1,314 patients with a primary tumor > 1.50 mm in thickness (T3-4N0M0; American Joint Committee on Cancer stage II) were randomly assigned to GM2-KLH/QS-21 vaccination (n = 657) or observation (n = 657). Treatment consisted of subcutaneous injections once per week from week 1 to 4, then every 3 months for the first 2 years and every 6 months during the third year. Primary end point was relapse-free survival (RFS). Secondary end points were distant metastasis-free survival (DMFS) and overall survival (OS). Analyses were by intent to treat.. After a median follow-up of 1.8 years, the trial was stopped at the second interim analysis for futility regarding RFS (hazard ratio [HR], 1.00; P = .99) and detrimental outcome regarding OS (HR, 1.66; P = .02). After a median follow-up of 4.2 years, we had recorded 400 relapses, nine deaths without relapse, a total of 236 deaths. At 4 years, the vaccination arm showed a decreased RFS rate of 1.2% (HR, 1.03; 95% CI, 0.84 to 1.25) and OS rate of 2.1% (HR, 1.16; 95% CI, 0.90 to 1.51). Toxicity was acceptable, with 4.6% of patients ending study participation because of toxicity.. GM2-KLH/QS-21 vaccination does not improve outcome for patients with stage II melanoma.

Topics: Adjuvants, Immunologic; Adult; Aged; Aged, 80 and over; Cancer Vaccines; Disease-Free Survival; Female; G(M2) Ganglioside; Hemocyanins; Humans; Kaplan-Meier Estimate; Lymph Nodes; Lymphatic Metastasis; Male; Melanoma; Middle Aged; Neoplasm Staging; Saponins; Skin Neoplasms; Watchful Waiting

High-dose interferon alfa-2b (IFNalpha2b) is the only established adjuvant therapy of resectable high-risk melanoma. GM2-KLH/QS-21 (GMK) is a chemically defined vaccine that is one of the best developed of a range of vaccine candidates for melanoma. A single-institution phase III trial conducted at Memorial Hospital served as the impetus for an intergroup adjuvant E1694/S9512/C509801 trial, which recently completed enrollment of 880 patients. To build on the apparent benefit of IFNalpha2b in resectable high-risk American Joint Committee on Cancer (AJCC) stage IIB or III melanoma, this phase II study was designed to evaluate the combination of GMK and IFNalpha2b. The E2696 trial was undertaken to evaluate the toxicity and other effects of the established adjuvant high-dose IFNalpha2b regimen in relation to immune responses to GMK and to evaluate the potential clinical and immunologic effects of the combined therapies.. This trial enrolled 107 patients with resectable high- or very high-risk melanoma (AJCC stages IIB, III, and IV).. The results demonstrate that IFNalpha2b does not significantly inhibit immunoglobulin M or G serologic responses to the vaccine and that the combination of high-dose IFNalpha2b and GMK is well tolerated in this patient population.. Cox analysis of the results of the combination with IFNalpha2b show improvement in the relapse-free survival of patients with very high-risk melanoma (including those with resectable M1 disease).

Topics: Adult; Antibody Formation; Antineoplastic Agents; Cancer Vaccines; Chemotherapy, Adjuvant; Combined Modality Therapy; Disease-Free Survival; Female; G(M2) Ganglioside; Humans; Interferon alpha-2; Interferon-alpha; Male; Melanoma; Middle Aged; Neoplasm Recurrence, Local; Recombinant Proteins; Risk Factors; Skin Neoplasms

β-1,4-N-Acetyl-Galactosaminyltransferase 1 (B4GALNT1) encodes the key enzyme B4GALNT1 to generate gangliosides GM2/GD2. GM2/GD2 gangliosides are surface glycolipids mainly found on brain neurons as well as peripheral nerves and skin melanocytes and are reported to exacerbate the malignant potential of melanomas. In order to elucidate the mechanism, we performed functional analyses of B4GALNT1-overexpressing cells. We analyzed ganglioside pattern on four melanoma and two neuroblastoma cell lines by high performance liquid chromatography (HPLC). We overexpressed B4GALNT1 in GM2/GD2-negative human melanoma cell line (SH4) and confirmed production of GM2/GD2 by HPLC. They showed higher anchorage independence growth (AIG) in colony formation assay, and exhibited augmented motility. In vitro, cell proliferation was not affected by GM2/GD2 expression. In vivo, GM2/GD2-positive SH4 clones showed significantly higher tumorigenesis in NOD/Scid/IL2Rγ-null mice, and immunostaining of mouse CD31 revealed that GM2/GD2 induced remarkable angiogenesis. No differences were seen in melanoma stem cell and Epithelial-Mesenchymal Transition markers between GM2/GD2-positive and -negative SH4 cells. We therefore concluded that B4GALNT1, and consequently GM2/GD2, enhanced tumorigenesis via induction of angiogenesis, AIG, and cell motility. RNA-Seq suggested periostin as a potential key factor for angiogenesis and AIG. These findings may lead to development of novel therapy for refractory melanoma.

Topics: Animals; Carcinogenesis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; G(M2) Ganglioside; Heterografts; Humans; Male; Melanoma; Mice; Mice, Inbred NOD; Mice, SCID; N-Acetylgalactosaminyltransferases; Neovascularization, Pathologic; Neuroblastoma; RNA-Seq; Skin Neoplasms; Transfection; Tumor Burden

Ganglioside functions in tumor metastasis were analyzed by carbohydrate remodeling of a mouse Lewis lung cancer (subline P29) by introducing beta1,4GalNAc-T cDNA. Although P29 was originally a low-metastatic subline in the s.c. injection system, it showed high potential in lung metastasis when i.v.-injected via the tail vein. Two lines of GM(2)(+) transfectants showed markedly reduced metastatic potential to the lung compared to 2 control lines. However, cell proliferation rates and expression levels of various cell adhesion molecules, e.g., integrin family members, SLe(x) and CD44, were essentially unchanged after transfection of the cDNA. Then, cell adhesion to fibronectin-coated dishes was examined, showing that GM(2) (+) transfectants attached to the plates much more slowly than controls, suggesting functional modulation of integrins with newly expressed GM(2). Phosphorylation of the FAK located at downstream of integrin molecules was markedly reduced in GM(2)(+) transfectants, suggesting that GM(2) suppressed cell adhesion signals via fibronectin-integrin interaction.

Topics: Animals; Carcinoma, Lewis Lung; Cell Membrane; DNA, Complementary; Female; Focal Adhesion Kinase 1; Focal Adhesion Protein-Tyrosine Kinases; G(M2) Ganglioside; Gangliosides; Integrins; Lung Neoplasms; Mice; Mice, Inbred C57BL; N-Acetylgalactosaminyltransferases; Neoplasm Transplantation; Phosphorylation; Protein-Tyrosine Kinases; Skin Neoplasms; Transfection; Tumor Cells, Cultured; Tyrosine; Vitronectin

An increased synthesis of the ganglioside GM2 has been reported on transformed murine cells, human fetal tissues, and transformed melanocytes. This study was designed to investigate whether any correlation existed between GM2 expression and the tumorigenicity of human melanoma. Ten established human melanoma cell lines, 5 rich in GM2 (group A) and 5 poor in GM2 (group B), were selected on the basis of previous ganglioside analysis of 28 melanoma cell lines. Six athymic nude mice per cell line were given an sc injection of 10(6) human melanoma cells/mouse. Tumors were measured every 2-4 days. By day 36 after the injection, 28 of 30 mice (93%) in group A developed medium to large tumors, but only 1 of 30 (3%) in group B developed a small tumor (P less than .005). The correlation of GM2 content of individual melanomas with tumor growth rate also was very high. GM2 content was expressed as nanomoles per gram wet weight of each melanoma. The area under the log values of tumor growth curves from day 0 to day 36, which represented tumor growth rate, tumor size, and latent period, was proportional to GM2 content, with a correlation coefficient of 0.927 and with P less than .001. The same log relationship when tested with other gangliosides, GM3, GD3, and GD2, was not statistically significant. These results, combined with the fact that variations in GM2 content do not affect in vitro growth of human melanoma cells, suggest that GM2 expression may be directly related to the dedifferentiation or the tumorigenicity of human melanoma.

Topics: Animals; Cell Differentiation; G(M2) Ganglioside; Gangliosides; Humans; Melanoma, Experimental; Mice; Mice, Nude; Neoplasm Transplantation; Skin Neoplasms