IN VIVO GENOTOXICITY TEST OF THE CHEMOPROPHYLACTIC ALBENDAZOLE USING THE SOMATIC MUTATION AND RECOMBINATION TEST (SMART)

• Grant number: 24/14580-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2025
• End date: November 30, 2025
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Luciana Pereira Silva
• Grantee: Marina Carbone
• Host Institution: Instituto Municipal de Ensino Superior de Assis. Fundação Educacional do Município de Assis (FEMA). Assis , SP, Brazil

Abstract

Benzimidazole derivatives, such as albendazole, can cause mutations and interfere with crucial cellular processes like mitosis, chromosome number, mitotic spindle function, and microtubule integrity in parasite larvae. There is evidence of its genotoxic potential in humans, including the induction of apoptosis, DNA fragmentation, and the production of reactive oxygen species, both in vitro and in vivo. Children treated with albendazole have shown an increase in sister chromatid exchanges and micronucleus formation in lymphocytes. Given this, the present project aims to evaluate the genotoxicity of albendazole using the Somatic Mutation and Recombination Test (SMART) in the wing cells of Drosophila melanogaster.To achieve this, crosses will be performed between two strains of D. melanogaster: flies with standard biometabolism and flies with high bioactivation capacity, allowing the generation of markers and balancers. From these crosses, transheterozygous marker individuals (MH) mwh/flr3, which will develop triplet hairs on the wings, and balancer heterozygous individuals (BH) mwh/TM3, BdS, which will develop flame-shaped hairs, differentiated by the wing edges, will be obtained.The larvae resulting from these crosses will be exposed to an aqueous solution containing albendazole suspension to verify drug-induced mutations in each subtype. The suspension will be diluted immediately before use in sterile distilled water, with standardized concentrations of 50%, 75%, and 100%. Third instar larvae from standard (ST) and high bioactivation (HB) crosses will be treated for approximately 48 hours with different concentrations of albendazole (ABZ), as well as with a negative control (distilled water) and a positive control (urethane 10 mM).After treatment, the survival rate of the larvae will be assessed by counting 100 individuals that reached hatching. After metamorphosis, adult flies emerging from the pupae will be collected, euthanized in an ethyl ether chamber, and preserved in 70% ethanol. The wings will be removed, fixed on slides, and analyzed under an optical microscope. Statistical analysis to verify genotoxicity will be based on the wing hair phenotype of treated samples and controls. The values will be analyzed using a two-tailed chi-square test for proportions, with a significance level of ¿ = 0.05. For mutation and recombination calculation, the following formula will be used: Mutation frequency (FM) = Frequency of mwh spots in BH offspring / Frequency of mwh spots in MH offspring; and Recombination = 1 - mutation frequency.