Effects of modulating WEE1 protein expression in Trypanosoma cruzi: a potential target for treatment

• Grant number: 24/23848-0
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: August 01, 2025
• End date: October 31, 2028
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Simone Guedes Calderano
• Grantee: Evelin Mariani Gonçalves
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Resistance to Chagas disease treatment with benznidazole (BNZ) is a growing concern, partly attributed to the ability of Trypanosoma cruzi amastigote forms to enter a state of dormancy. This state allows the parasite to persist and remain infective even after treatment cessation. Dormancy may occur spontaneously or be induced by BNZ itself, which causes DNA damage and activates repair mechanisms mediated by cell cycle checkpoints. Despite its relevance, these processes remain poorly understood in trypanosomatids, especially regarding checkpoint regulation. The presence of WEE1, a regulatory kinase known for inhibiting CDKs in response to DNA damage, may be a key factor in this process. This protein, whose function is well-established in mammals and yeast, inhibits cell cycle progression, primarily at the G2/M checkpoint, contributing to genomic integrity maintenance. In preclinical cancer studies, WEE1 inhibition combined with chemo/radiotherapeutic agents has been shown to increase tumor cell sensitivity to treatment, leading to irreparable genomic damage and, ultimately, apoptosis. However, the role of WEE1 in trypanosomatids like T. cruzi remains largely unexplored. In T. cruzi, the cell cycle is regulated by kinases homologous to CDK1, known as Cdc2 Related Kinases (CRKs), which interact with cyclins to promote cell cycle progression. Although specific inhibitors for CRKs have yet to be identified, WEE1 is present in the T. cruzi genome, with conservation of the phosphorylation site in several CRKs, suggesting a potential regulatory function of these kinases. Studies conducted by our research group have revealed that WEE1 exhibits variable expression during the cell cycle and metacyclogenesis, along with nuclear localization, suggesting its involvement in these processes. However, it remains unclear whether WEE1 plays a role in these parasites similar to that observed in other eukaryotes, particularly in regulating specific checkpoints. This project aims to investigate the effects of modulating WEE1 expression in dormant and resistant amastigotes. Furthermore, it seeks to explore whether WEE1 inhibition can sensitize the parasite to BNZ, as observed in oncological studies, offering a novel therapeutic perspective for Chagas disease.