Molecular and morphological parameters for monitoring the Drug Holiday protocol in 3D culture

• Grant number: 22/15626-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: June 01, 2023
• End date: September 01, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Carmen Veríssima Ferreira
• Grantee: Beatriz Aires Lopes
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated scholarship(s): 24/05180-1 - Tracking the infiltration capacity of NK cells in breast cancer spheroids after violacein treatment, BE.EP.MS

Abstract

Melanoma is defined as an aggressive type of cancer due to its high ability to develop metastases and low response to therapy. In recent decades its incidence has been increasing and by 2040, estimates predict an increase of 50% in the number of cases and 68% in the number of deaths. Usually the melanoma studies are based on 2D cell culture, however this model is not able to mimic important characteristics of the tumor in vivo, which promote the low response rate in this disease. In this sense, the use of 3D cell cultures provides greater fidelity to the tumor physiology present in the body. Regarding the molecular mechanisms that promote such aggressiveness, the Fatty Acid Synthase (FASN) is important for membrane turnover and de novo synthesis, which has a role in the autophagosome formation, a step of the autophagic process (one of the main mechanisms of resistance in melanoma). Tyrosine protein kinase receptor, AXL, is related to the activation of pro-tumorigenic signaling pathways which promotes migration and invasion. In addition, recent evidence has also associated it with the autophagic process. Our study demonstrated that violacein, a secondary metabolite derived from Chromobacterium violaceum, found in the Amazon River, modulates the expression of such proteins in 2D cell culture and in spheroids (3D). However, despite the relevant findings, considering the rapid tumor plasticity of melanoma, in the present project we will focus on: I. Establishing a long-term culture of spheroids, aimed to track responses to treatment for a longer period of time; II. Evaluating the response of spheroids to treatment with the ViVe protocol (violacein + vemurafenib) and Drug Holiday therapeutic scheme, evaluating the morphological and molecular responses, through the expression and location of AXL, FASN and p62 tracking.