EFFECTS OF iNOS AND METABOLIC INHIBITION IN HUMAN PRIMARY GLIOMA CULTURES

HINCAPIÉ ARIAS EL ; BELGOROSKY D ; MERENZON M ; SEOANE ER ; YASUDA E ; RECALDE R ; SOKOLOVSKY M ; ZANINOVICH R ; VILLAVERDE M ; EIJAN ANA MARIA

MAR DEL PLATA

Congreso; Reunion Anual de Sociedades de Biociencias SAIC-SAI-SAFIS; 2022

Introduction: Gliomas (GM) are a group with varied cellular origins, from low to high grade (I-IV, OMS). Despite treatment with surgery/radiotherapy/chemotherapy, malignant and some benign gliomas in eloquent areas or of vital importance are inoperable and/or have a dismal prognosis. It has been suggested that inducible nitric oxide synthase isoform enzyme (iNOS) is related to glioma growth, progression and cancer stem cell (CSC) maintenance; and that the selective inhibition of glycolytic pathways is related to a decreased proliferation of glioma cells.Objective: Evaluate the effect of iNOS specific inhibitor, S-methylisothiourea (SMT); and the effect of glycolytic inhibitor, 2-Deoxy-D-glucose (2DG), alone or in combination with TMZ, on 2D (monolayer) and on the glioma stem cell (GSC) niche. Methodology: Endorsement was obtained from the ethics committees of 3 hospitals (Ángel H. Roffo, Clínicas, Ramos-Mejía); and signed informed consents. Samples were obtained between 2018 and 2022 from the operating room and immediately processed by mechanical disintegration with DMEM-F12 and 10% fetal bovine serum; seeded in 2D and under sphere conditions (low adhesion and high dilution). Cell viability was determined in 2D by MTS. The number of CSC was established by sphere forming efficiency (SFE) in relation to the number of seeded cells and their diameters were measured with imageJ.Results: 11 human GM samples were processed. 6 cultures prospered with at least 1 cell passage. In 2D, 2DG decreased cell viability (50 - 70%); SMT has no significant effect. In the GSC niche, SMT alone decreased SFE (40 - 60%, p