Establishment of a low-cost 3D tri-culture model to study the effects of emerging contaminants on fatty liver-inducing dysfunctions and functional validation of the ChREBP protein activity

• Grant number: 25/01570-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: July 01, 2025
• End date: January 31, 2029
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal Investigator: Karen Cristiane Martinez de Moraes
• Grantee: Joyce Alessandra Lima
• Host Institution: Instituto de Biociências (IB). Universidade Estadual Paulista (UNESP). Campus de Rio Claro. Rio Claro , SP, Brazil

Abstract

Nonalcoholic fatty liver diseases (NAFLD) are currently considered a global public health issue. Presenting different etiologies, these pathologies are characterized by an excessive accumulation of fat in the liver, which may progress to more sever clinical conditions. In our society, the excessive use of 2 chemical products, such as pesticides, has significantly contributed to environmental contamination, affecting drinking water sources worldwide and exposing non-target animals to these compounds, which may contribute to increased number of cases of liver and metabolic dysfunctions. Thus, the development of an alternative methodology to the use of animals in research is considered a valuable tool in predictive toxicology to evaluate the adverse effects of contaminants. Considering that in previous studies carried out in our laboratory, where we developed a low-cost 3D co-culture of liver cells to investigate pro steatotic environment, in the present scientific proposal we aimed to establish a 3D triculture to serve as a biotechnological tool for the study of hepatic metabolic dysfunctions element. The 3D triculture will be exposed to the pesticides s-metolachlor and chloratalonil and the concentrations evaluated will be those acceptable by regulatory agencies. These pesticides are often found in drinking waters of different countries. Next, cellular, molecular and biochemical analyses will be performed to evaluate the adverse signaling routes that facilitate the establishment of pro-steatotic environment. In addition, we will evaluate the functional role of the ChREBP protein, an important element in hepatic metabolism, whose understanding of its functionality against pesticide poisoning is still required. For the mechanistic analysis of the ChREBP genetic tools and/or chemical blockers of the functional activity of the protein will be used and the ChREBP-modified spheroids will be exposed to pesticides in study. Following analysis of RNA-seq will elucidate elements of fine regulation of ChREBP in toxic environments, aiming at characterization of markers and biotechnological applications. (AU)