Effects of the combination of coffee compounds and antitumor agents on Hepatocellular Carcinoma: in vitro and in vivo investigation

• Grant number: 23/05411-0
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: November 01, 2023
• End date: February 28, 2027
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Guilherme Ribeiro Romualdo
• Grantee: Leticia Cardoso Valente
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The Hepatocellular Carcinoma (HCC) displays a high incidence and mortality worldwide, also featuring a poor prognosis. Many patients in advanced disease stages are submitted to low response antitumoral therapies. Tumor microenvironment modulation is also neglected in current targeted therapies. A myriad of epidemiological studies indicate that coffee consumption reduced by 40% the HCC risk. This beverage is a reservoir of several bioactive compounds (BCs) that have already demonstrated antiproliferative, pro-apoptotic and anti-inflammatory effects in preclinical studies, such as caffeine (CAF), chlorogenic acid (ACG), kahweol (KWL). Due to the need to increase the effectiveness of current therapies, we will assess whether the administration - alone or in combination - of the CAF alkaloid, the diterpene KWL and the polyphenol ACG promotes the antitumoral effects of Sorafenib therapy or immunotherapy by Atezolizumab+Bevacizumab in in vitro and in vivo models of HCC. Co-culture 3D models of HCC tumor cells (HepG2) and stellate cells (LX2) will be established. Co-cultures will be treated with BCs - alone or in combination - and with the respective therapies for 24 or 48 h. Cell viability (MTT), cytotoxicity (LDH), motility (scratch assay), and immunoexpression of Ki-67 (proliferation), caspase-3 (apoptosis) and VEGF (angiogenesis) will be evaluated. Then, the Combinatorial Index (CI) will be calculated and the combinations with additive/synergistic results will be selected for the in vivo study. BALB/c nude mice will be subjected to xenograft tumor model by the inoculation of HepG2/LX2 cells. The animals will be treated with the selected BCs and respective therapies, and the immunoexpression of Ki-67, caspase-3, CD31 and VEGF, and the main tumorigenic pathways and tumor microenvironment at the transcriptomic level (NanoString) will be evaluated in tumors. The main altered pathways will be validated at the protein level (immunohistochemistry and immunoblot) in human samples, using Patient-derived xenograft methodology. HCC human samples of will be collected, and implanted in immunosuppressed mice, and treated with selected therapies and BCs. (AU)

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)

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)