Modulation of Trypanosoma cruzi infection in HTR-8/SVneo cells by Copaifera spp. oleoresins and isolated compound (ent-kaurenoic acid)

Abstract

Introduction: Congenital Chagas Disease (CCD), caused by Trypanosoma cruzi, is a major public health issue and the main form of infection of Chagas Disease (CD) in non-endemic countries. While CD is treated with nifurtimox and benznidazole, these are not recommended for congenital and gestational CD due to limited fetal safety data, highlighting the urgent need for alternatives. Copaifera spp. oleoresins are known for microbicidal effects, including anti-T. cruzi activity, but few studies address their impact on the maternal-fetal interface. Objectives: This study evaluated three Copaifera oleoresins and one isolated compound for modulating T. cruzi infection in human extravillous trophoblastic cells (HTR-8/SVneo). Methods: Cell viability was assessed by MTT; T. cruzi proliferation and invasion by parasite counting; cytokines by ELISA; horizontal cell migration and cell invasion by Wound Healing and Transwell assays, respectively; and in silico analysis by molecular docking. Results: The oleoresins and ent-kaurenoic acid significantly reduced amastigote proliferation, while only ent-kaurenoic acid controlled parasite invasion in HTR-8/SVneo cells. Cytokine analysis showed downregulation of IFN-γ, TNF-α, IL-8, TGF-β1 and IL-10, indicating maintained immune tolerance needed in pregnancy. The migration assay showed all treatments restored normal migration in HTR-8/SVneo cells, which was impaired by infection. However, the Transwell assay showed oleoresins and ent-kaurenoic acid did not restore impaired cell invasion by T. cruzi. Finally, molecular docking showed ent-kaurenoic acid had the best binding to T. cruzi glyceraldehyde-3-phosphate dehydrogenase, suggesting a potent parasite target. Conclusion: These findings offer important insights for therapies against T. cruzi infection at the human maternal-fetal interface.