The antischistosomal potential of GSK-J4, an H3K27 demethylase inhibitor: insights from molecular modeling, transcriptomics and in vitro assays

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Lobo-Silva, Jessica ^[1] ; Cabral, Fernanda J. ^[2] ; Amaral, Murilo S. ^[3] ; Miyasato, Patricia A. ^[4] ; de Freitas, Rafaela Paula ^[4] ; Pereira, Adriana S. A. ^{[5, 3]} ; Khouri, I, Mariana ; Barbosa, Mayra M. F. ^[6] ; Ramos, Pablo I. P. ^[7] ; Leite, Luciana C. C. ^[6] ; Asojo, Oluwatoyin A. ^[8] ; Nakano, Eliana ^[4] ; Verjovski-Almeida, Sergio ^{[5, 3]} ; Farias, Leonardo P. ^[9]

Afiliação do(s) autor(es)

^[1] Fundacao Oswaldo Cruz, Lab Biomarcadores & Inflamacao, Inst Goncalo Moniz, Salvador, BA - Brazil

^[2] Univ Estadual Campinas, Inst Biol, Dept Biol Anim, Campinas, SP - Brazil

^[3] Inst Butantan, Lab Expressao Gen Eucariotos, Sao Paulo, SP - Brazil

^[4] Inst Butantan, Lab Parasitol, Sao Paulo, SP - Brazil

^[5] Univ Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo - Brazil

^[6] Inst Butantan, Lab Especial Desenvolvimento Vacinas, Sao Paulo, SP - Brazil

^[7] Fundacao Oswaldo Cruz, Inst Goncalo Moniz, Ctr Integracao Dados & Conhecimentos Saude CIDACS, Salvador, BA - Brazil

^[8] Hampton Univ, Dept Chem & Biochem, Hampton, VA 23668 - USA

^[9] Khouri, Mariana, I, Fundacao Oswaldo Cruz, Lab Biomarcadores & Inflamacao, Inst Goncalo Moniz, Salvador, BA - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PARASITES & VECTORS; v. 13, n. 1 MAR 17 2020.
• Citações Web of Science: 0

Resumo

Background Schistosomiasis chemotherapy is largely based on praziquantel (PZQ). Although PZQ is very safe and tolerable, it does not prevent reinfection and emerging resistance is a primary concern. Recent studies have shown that the targeting of epigenetic machinery in Schistosoma mansoni may result in severe alterations in parasite development, leading to death. This new route for drug discovery in schistosomiasis has focused on classes of histone deacetylases (HDACs) and histone acetyltransferases (HATs) as epigenetic drug targets. Schistosoma histone demethylases also seem to be important in the transition of cercariae into schistosomula, as well as sexual differentiation in adult worms. Methods The Target-Pathogen database and molecular docking assays were used to prioritize the druggability of S. mansoni histone demethylases. The transcription profile of Smp\_03400 was re-analyzed using available databases. The effect of GSK-J4 inhibitor in schistosomula and adult worms' motility/viability/oviposition was assessed by in vitro assays. Ultrastructural analysis was performed on adult worms exposed to GSK-J4 by scanning electron microscopy, while internal structures and muscle fiber integrity was investigated by confocal microscopy after Langeron \& apos;s carmine or phalloidin staining. Results The present evaluation of the potential druggability of 14 annotated S. mansoni demethylase enzymes identified the S. mansoni ortholog of human KDM6A/UTX (Smp\_034000) as the most suitable druggable target. In silico analysis and molecular modeling indicated the potential for cofactor displacement by the chemical probe GSK-J4. Our re-analysis of transcriptomic data revealed that Smp\_034000 expression peaks at 24 h in newly transformed schistosomula and 5-week-old adult worms. Moreover, this gene was highly expressed in the testes of mature male worms compared to the rest of the parasite body. In in vitro schistosome cultures, treatment with GSK-J4 produced striking effects on schistosomula mortality and adult worm motility and mortality, as well as egg oviposition, in a dose- and time-dependent manner. Unexpectedly, western blot assays did not demonstrate overall modulation of H3K27me3 levels in response to GSK-J4. Confocal and scanning electron microscopy revealed the loss of original features in muscle fibers and alterations in cell-cell contact following GSK-J4 treatment. Conclusions GSK-J4 presents promising potential for antischistosomal control; however, the underlying mechanisms warrant further investigation. (AU)

• Processo FAPESP: 15/06366-2 - Avaliando os mecanismos de auto-cura em macacos rhesus infectados com Schistosoma mansoni como uma nova base para uma vacina
• Beneficiário: Sergio Verjovski Almeida
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 17/07364-9 - Estudo das modificações pós-traducionais de histonas e de co-reguladores da expressão nos estágios de vida do Schistosoma mansoni
• Beneficiário: Fernanda Janku Cabral
• Modalidade de apoio: Auxílio à Pesquisa - Regular