Gene Expression and Genetic Polymorphism of PNPLA3 in Metabolic Dysfunction-Associated Steatotic Liver Disease and Its Relationship with Hepatocellular Carcinoma

• Grant number: 24/11492-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2024
• End date: May 31, 2025
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Dorotéia Rossi Silva Souza
• Grantee: Lorena Alves Oliveira da Silva
• Host Institution: Faculdade de Medicina de São José do Rio Preto (FAMERP). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São José do Rio Preto , SP, Brazil

Abstract

Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD), characterized by the accumulation of triglycerides in hepatocytes, is a manifestation of metabolic syndrome and has an increasing incidence worldwide. This condition predisposes individuals to the development of fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Among the events preceding carcinogenesis in HCC, the inflammatory microenvironment and fibrosis, induced by the accumulation of triglycerides in liver cells, stand out. Genetic variants have been associated with MASLD progression and the development of HCC. In this context, PNPLA3 (patatin-like phospholipase domain-containing), a membrane lipase (adiponutrin), plays a role in hepatic triglyceride metabolism. Analyzing molecular markers related to metabolic pathways may clarify mechanisms involved in MASLD-associated hepatic carcinogenesis and contribute to the diagnosis and prognosis of the disease. This study aims to analyze the gene expression of PNPLA3 in tissue from patients with MASLD with and without HCC, and its association with PNPLA3 genetic polymorphism (rs738409), as well as survival and histopathological characteristics, including tissue infiltration, metastasis, and cellular differentiation. A total of 110 individuals will be studied, distributed as follows: 30 patients with HCC, 30 patients with MASLD, 30 patients with MASLD and HCC, and 20 individuals without clinical signs of liver diseases (control group - liver tissue samples from autopsies). Formalin-fixed, paraffin-embedded tissue samples will be used for RNA and DNA extraction, for gene expression analysis and genotyping, respectively, by real-time polymerase chain reaction. Clinical data will be obtained from electronic medical records. Data will be analyzed statistically, including chi-square (Dz) or Fisher's test for qualitative variables, analysis of variance (ANOVA) and t-test, or Kruskal-Wallis and Mann-Whitney tests for quantitative variables with or without Gaussian distribution, respectively, and Kaplan-Meier curve for survival analysis. An alpha error of 5% will be considered acceptable.