Evaluation of JAK-STAT5 and mTOR-S6 signaling pathways in functional modulation of natural killer (NK) cells from patients with epithelial ovarian cancer

• Grant number: 18/06959-1
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2018
• End date: March 31, 2021
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: Fernando Guimarães
• Grantee: Fernando Guimarães
• Host Institution: Centro de Atenção Integral à Saúde da Mulher (CAISM). Hospital da Mulher Professor Doutor José Aristodemo Pinotti. Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated researchers: Daniela Maira Cardozo ; Fernando Souza-Fonseca-Guimaraes ; José Andrés Yunes ; Nicholas David Huntington ; Rodrigo Fernandes da Silva ; Sophie Françoise Mauricette Derchain
• Associated scholarship(s): 20/11628-4 - Cell characterization of peripheral blood mononuclear cell samples and ascites cells of patients with advanced ovarian epithelial cancer., BP.TT; 19/25113-9 - Cell characterization of peripheral blood mononuclear cell samples and ascites cells of patients with advanced ovarian epithelial cancer, BP.TT; 18/18047-7 - Cellular characterization of peripheral blood mononuclear cells and ascites cells from patients with advanced ovarian epithelial cancer, BP.TT

Abstract

Natural killer lymphocytes (NK) are known for the ability to kill neoplastic cells and fight the metastatic spread of cancer. Metastatic dissemination is the main cause of death associated with cancer, especially in advanced epithelial ovarian cancer (EOC). We have recently shown that functional performance of NK cells is significantly higher in EOC associated-ascites without malignant cells compared with EOC ascites with carcinomatosis or even, compared with NK cells from heathy women's blood. Additionally, NK cells from ascites with carcinomatosis are unresponsive to interleukin (IL)-2 stimuli. The JAK-STAT5 and mTOR-S6 intracellular pathways have been pointed as pivotal in the convergence of signals delivered by the activating cytokines IL-2 and Il-15 to NK cells and, therefore, implicated in the functional modulation of these cells. Thus, this study aims to evaluate the role of JAK-STAT5 and mTOR-S6 pathways in relation to the functional modulation of NK cells from patients with advanced EOC. This study will include 35 women assisted at the Woman's Hospital of the University of Campinas for treatment of EOC, whose clinical examination and ultrasonography indicate the presence of peritoneal ascites. These women will be subjected to peripheral blood and ascites collection (observing clinical indication for the procedures). Subjects without EOC confirmation by anatomopathological analysis will be excluded from the study. NK cells from blood and ascites will be assessed for their function, phenotype, STAT5 and S6 protein phosphorylation, integrity and responsiveness of cytokines receptors IL-2 and IL-15, as well as, expression of genes that codify proteins of JAK-STAT5 and mTOR-S6 pathways. Ascites will be further assessed for the presence of malignant cells, lymphocytes subtypes and cytokines profile. In vitro cocultivation assays with CAISMOV24 cells (EOC cell lineage) will be employed to assess the possible immunomodulatory mechanisms of NK cells. Descriptive analysis of the data will be performed, as well as comparisons between variables with ANOVA followed by post hoc test. Stepwise linear regression will be used to evaluate the influence of variables such as the intensity of phosphorylation of STAT5 and S6 proteins, expression of activating receptors, presence of regulatory T lymphocytes and others in NK cell function. The study will count on the collaboration of two researchers from "The Walter and Eliza Hall Institute of Medical Research" (Melbourne, Australia), where a similar study has being conducted. Collaboration includes exchange of information and analysis in bioinformatics. The JAK-STAT5 and mTOR-S6 pathways have been implicated in the functional modulation of NK cells and in their ability to control tumor growth and metastatic spread. Thus, this study will expand the knowledge about the potential use of these signaling pathways as therapeutic targets in the treatment of the EOC. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

TONETTI, CLAUDIA RODRIGUES; DE SOUZA-ARAUJO, CAROLINE NATANIA; YOSHIDA, ADRIANA; DA SILVA, RODRIGO FERNANDES; ALVES, PAULO CESAR MARTINS; MAZZOLA, TAIS NITSCH; DERCHAIN, SOPHIE; FERNANDES, LUIS GUSTAVO ROMANI; GUIMARAES, FERNANDO. Ovarian Cancer-Associated Ascites Have High Proportions of Cytokine-Responsive CD56bright NK Cells. CELLS, v. 10, n. 7, JUL 2021. (18/18047-7, 19/25113-9, 20/11628-4, 18/06959-1)

DE SOUZA-ARAUJO, CAROLINE NATANIA; TONETTI, CLAUDIA RODRIGUES; CARDOSO, MARCELLA REGINA; LUCCI DE ANGELO ANDRADE, LILIANA APARECIDA; DA SILVA, RODRIGO FERNANDES; ROMANI FERNANDES, LUIS GUSTAVO; GUIMARAES, FERNANDO. Three-Dimensional Cell Culture Based on Magnetic Fields to Assemble Low-Grade Ovarian Carcinoma Cell Aggregates Containing Lymphocytes. CELLS, v. 9, n. 3, MAR 2020. (18/06959-1)