Effects of carboxymethylcellulose and polysorbate 80 food additive on hepatocarcinogenesis-associated non-alcoholic fatty liver disease.

• Grant number: 22/13402-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: April 01, 2023
• Status: Discontinued
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Luís Fernando Barbisan
• Grantee: Gabriel Bacil Prata
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated scholarship(s): 24/17423-6 - Early obesity and hepatocellular carcinoma: unveiling the adipose tissue and liver carcinogenesis interplay, BE.EP.DR

Abstract

The hepatocellular carcinoma (HCC) features as the 6th/3rd most incident and deadliest, respectively, among all kind of cancers worldwide, and it has been linked to chronic liver diseases like non-alcoholic fatty liver disease (NAFLD). Nowadays, NAFLD is the most common cause of chronic liver disease worldwide and affects almost 25% of the global population, also contributing for increasing the HCC-related incidence by 26%. Moreover, the adherence to a western diet (WD) - mostly composed by saturated fatty acids, sugars, and food additive - has been targeted as driver of NAFLD, also increasing the risk of developing HCC by 80%. The ultra-processed food (UPF) is characterized its hypercaloric and food additive-enriched profile (eg.: sugars, emulsifiers, flavourings, and dyes), consisting of ~60% of the suggested daily-intake calories. Among the most common used emulsifiers additives, carboxymethylcellulose (CMC) and polysorbate 80 (P80) shows potential effects on the liver-gut-adipose axis, modulating the hepatic damage levels, microbiome profile, gut permeability, adipogenesis, and glucose/insulin dynamics, also prospecting as potential drivers of HCC, although there are not preclinical findings about it. We sought to assess the effects of CMC and/or P80 on both in vivo and in vitro models of HCC-associated NAFLD. Male C57BL/6 were distributed into 11 experimental groups (G1-G11) and received intraperitoneal injections of diethylnitrosamine [DEN, 25mg/Kg of body weight (b.w.), 1x/week, for 4 weeks] or vehicle, starting at the 14th post-natal day. At the 6th week of life, mice also received a WD (hypercaloric chow: 30% of saturated fat/20% of sucrose/0.2% of cholesterol; and a high-sugar solution: 55/45% of d-frcutose/d-glucose, for drinking) or a basal diet, for further 24 weeks. Simultaneously, mice received intragastric injections of CMC (370.0 or 740.0 mg/Kg of b.w., 5x/week, for 24 weeks) and/or P80 (100.0 or 200.0 mg/Kg of b.w., 5x/week, for 24 weeks) and were euthanized at the end of 24th week of protocol. Neoplastic and hepatic (histopathological, lipid/collagen content, immunohistochemistry, metabolomic, and mRNAseq), adipose tissue (histopathological and immunohistochemistry), small intestine (immunohistochemistry, toluidine blue and mucin density), feces (microbiome profiling), and serum (glucose tolerance test and determination of alanine aminotransferases, total cholesterol, and triglycerides levels) samples will be further assessed. In a co-culture spheroid model added to a fatty acid-supplemented medium, cell viability and lipid/collagen content will be assessed. Data will be analyzed by one-way ANOVA or Kruskal-Wallis and Tukey pos hoc test and will be considered statistically different when p<0.05.

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)

ROMUALDO, GUILHERME RIBEIRO; HEIDOR, RENATO; BACIL, GABRIEL PRATA; MORENO, FERNANDO SALVADOR; BARBISAN, LUIS FERNANDO. Past, present, and future of chemically induced hepatocarcinogenesis rodent models: Perspectives concerning classic and new cancer hallmarks. Life Sciences, v. 330, p. 16-pg., 2023-10-01. (23/08751-7, 22/16633-1, 22/13402-9, 22/06082-8)

VALENTE, LETICIA CARDOSO; BACIL, GABRIEL PRATA; RIECHELMANN-CASARIN, LUANA; BARBOSA, GIULLIA CAVICHIOLLI; BARBISAN, LUIS FERNANDO; ROMUALDO, GUILHERME RIBEIRO. Exploring in vitro modeling in hepatocarcinogenesis research: morphological and molecular features and similarities to the corresponding human disease. Life Sciences, v. 351, p. 13-pg., 2024-06-13. (22/13402-9, 23/05411-0, 23/08751-7, 22/16633-1, 23/17585-3)