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Influence of variants in genes of immunological pathways in risk and prognosis of patients with cutaneous melanoma

Grant number: 19/09168-8
Support type:Regular Research Grants
Duration: November 01, 2019 - October 31, 2021
Field of knowledge:Health Sciences - Medicine - Medical Clinics
Principal Investigator:Carmen Silvia Passos Lima
Grantee:Carmen Silvia Passos Lima
Home Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Assoc. researchers:Gustavo Jacob Lourenço


Cutaneous melanoma (CM) deserves prominence among tumors due to its high mortality. The success of immunotherapy in patients with CM indicates that the immune system acts in the control of the tumor. Recently, single-base genetic variants (SNVs) in the PD1 pathway were associated with our group with the risk and prognosis of CM. It is quite possible that SNVs in genes of other pathways of the immune system have similar or greater importance in the origin and progression of the tumor. We intend to evaluate in this study the association of SNVs in gene TNFRSF1B (c.*215C>T, c.*188A>G, c.*922C>T and c.587T>G) of the tumor necrosis factor receptor 2 (TNFR2) pathway, HAVCR2 (c.-574A>C, c.-882G>A, c.-1766C>A and c.*931A>G) of the T cell immunoglobulin and mucin-domain containing-3 (TIM-3) pathway and JAK1 (c.1648+1272G>A, c.991-27C>T), JAK2 (c.-1132G>T, c.-139G>A) and STAT3 (c.*1671T>C, c.-1937C>G) of janus kinase signal transducer of activation (JAK/STAT) pathway of the immune system with the risk of occurrence, clinicopathological aspects, and prognosis of CM patients. Three-hundred patients with CM and 300 blood donors will be evaluated at the Institution. Clinical information will be obtained through specific questionnaires and access to medical records. Genotypes will be identified by real-time polymerase chain reaction (PCR) in genomic DNA. The expression of the gene will be analyzed by quantitative PCR in the genomic RNA of individuals with the different genotypes of each SNV. The functional roles of SNVs of greater interest (the one with the most consistent association with risk, clinicopathological aspects or patient survival) for each of the TNFRSF1B, HAVCR2, and STAT3 genes will be evaluated. If the SNV of interest is located in an intronic region, the reporter minigene assay will be performed and if in the promoter region or 3'-UTR region, a luciferase assay will be performed in a genetically modified SK-MEL-28 melanoma cell line to present the ancestral and variant genotypes of the SNV of interest; flow cytometry to assess the cell cycle and apoptosis will complement those assays. Differences between groups will be assessed by the Fisher or chi-square test. Comparisons of gene and protein expression will be performed using t-tests and ANOVA or Mann-Whitney and Kruskal-Wallis tests. Survival analyses will be performed using the Kaplan-Meier method, log-rank test and Cox's univariate and multivariate analyses. The results of this study may contribute to identifying individuals at high risk for CM or aggressive melanoma, who deserve to receive measures for early diagnosis and/or differentiated therapy. (AU)