KRASgetaway: a preclinical investigation for control of the KRAS oncogene

• Grant number: 23/02032-9
• Support Opportunities: Regular Research Grants
• Start date: September 01, 2023
• End date: November 30, 2025
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: Renato José da Silva Oliveira
• Grantee: Renato José da Silva Oliveira
• Host Institution: Hospital do Câncer de Barretos. Fundação Pio XII (FP). Barretos , SP, Brazil
• Associated researchers: Ana Carolina Laus ; Josiane Mourão Dias ; Letícia Ferro Leal ; Olga Catarina Lopes Martinho ; Rui Manuel Vieira Reis ; Silvia Aparecida Teixeira

Abstract

Gain-of-function mutations in the KRAS gene lead to constitutive protein activity, which is associated with tumorigenesis and tumor progression in pancreatic, colorectal, and lung cancer. The challenge in creating therapies against KRAS has extended for decades due to the structural complexity of this protein. However, after the discovery of druggable portions in some of the KRAS variants, it became possible to design inhibitors with improved potency, selectivity, and bioavailability such as Sotorasib for the G12C variant, and MRTX1133 for the G12D variant. However, the molecular mechanisms involved in resistance and failure represent the greatest current challenge for this treatment and need to be understood. The objective of the project is to preclinically investigate the molecular alterations that lead to failure in response to approved (anti-KRAS G12C) and late-stage approved (anti-KRAS G12D) KRAS therapies, seeking a therapeutic solution for acquired resistance to these inhibitors. The project will be divided into three stages: (1) Creation of resistance models to KRAS inhibitors; (2) Search for markers of resistance to anti-KRAS therapies; and (3) Evaluation of therapeutic combinations capable of restoring sensitivity to KRAS inhibitors. A total of 4 mutant KRAS tumor cell lines (G12C or G12D) will be continuously exposed to KRAS inhibitors (Sotorasib and MRTX1133) to create resistant clones, where molecular changes in the expression profile of genes related to proliferation and immune response pathways will be evaluated by the NanoString platform. Additionally, differential protein expression of resistant clones relative to sensitive ones will be evaluated by protein arrays of KRAS-mediated signaling pathways and immune response protein. In vitro molecular findings will be validated in tumor samples from patient's refractory to anti-KRAS therapy treated at this institution. Therapeutic combinations will be proposed to control resistance to KRAS inhibitors, evaluated through cellular assays of proliferation, invasion, migration, colony formation, analysis of cell cycle and cell death in 2D and 3D cellular models, and xenograft models. We hope that this preclinical investigation platform can identify biomarkers related to anti-KRAS therapy failure and demonstrate the best therapeutic combinations to control the KRAS oncogene in patients still without treatment prospects, after progression disease. (AU)

Scientific publications

(4)

(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)

STRINGHETTA, GIULIA RODRIGUES; MASS, EDUARDO BUSTOS; GOMES, IZABELA NATALIA FARIA; PEIXOTO, MARIA CLARA FONSECA; TEJADA, AMANDA HELENA; SUSUCCHI, LUCIANE; BEZERRA, ARYEL JOSE ALVES; RESENDE, PEDRO VICTOR SILVA; VENDRUSCULO, VINICIUS; REIS, MANUEL; et al. Design, synthesis and evaluation of quinazoline-chalcone hybrids as inducers of cell-cycle arrest and apoptosis in breast cancer via DNA damage and CDK2/ATR inhibition. EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY REPORTS, v. 13, p. 16-pg., 2025-04-01. (23/02032-9, 22/05597-4)

RESENDE, PEDRO VICTOR SILVA; GOMESA, IZABELA NATALIA FARIA; PEIXOTO, MARIA CLARA; STRINGHETTA, GIULIA RODRIGUES; ARANTES, LIDIA MARIA REBOLHO BATISTA; KUZMIN, VLADIMIR ALEXANDROVICH; BORISSEVITCH, IOURI; REIS, RUI MANUEL; VAZQUEZA, VINICIUS DE LIMA; FERREIRA, LUCIMARA PERPETUA; et al. Evaluation of the antineoplastic properties of the photosensitizer biscyanine in 2D and 3D tumor cell models and artificial skin models. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY, v. 262, p. 10-pg., 2024-12-12. (23/02032-9, 22/05597-4)

MEGID, RAQUEL ARANTES; RIBEIRO, GUILHERME GOMES; GOMES, IZABELA NATALIA FARIA; LAUS, ANA CAROLINA; LEAL, LETICIA FERRO; DA SILVA, LUCIANE SUSSUCHI; ARIWOOLA, ABU-BAKR ADETAYO; DIAS, JOSIANE MOURAO; REIS, RUI MANUEL; DA SILVA-OLIVEIRA, RENATO JOSE. Sotorasib resistance triggers epithelial-mesenchymal transition and activates AKT and P38-mediated signaling. FRONTIERS IN MOLECULAR BIOSCIENCES, v. 12, p. 14-pg., 2025-01-30. (23/02032-9)

LAUS, ANA CAROLINA; GOMES, IZABELA NATALIA FARIA; DA SILVA, ALINE LARISSA VIRGINIO; DA SILVA, LUCIANE SUSSUCHI; MILAN, MIRELLA BARONI; APARECIDATEIXEIRA, SILVIA; MARTIN, ANA CAROLINA BAPTISTA MORENO; PEREIRA, LETICIA DO NASCIMENTO BRAGA; DE CARVALHO, CARLOS EDUARDO BARBOSA; CROVADOR, CAMILA SOUZA; et al. Establishment and molecular characterization of HCB-541, a novel and aggressive human cutaneous squamous cell carcinoma cell line. HUMAN CELL, v. 37, n. 4, p. 14-pg., 2024-04-03. (23/02032-9, 17/22305-9)