Authors
Mariza Gabriela Faleiro de Moura Lodi Cruz, Fernanda Viana Moreira Silva, Giovanna Cecília de Paula E Sá, Juliana Martins Ribeiro, Giovanna Ferreira da Fonseca, Davi Alvarenga Lima, Karine Ferreira Lopes, Ana Maria Murta Santi, Silvane Maria Fonseca Murta
Published in
Antimicrobial agents and chemotherapy. Pages e0024726. Jul 06, 2026. Epub Jul 06, 2026.
Abstract
Leishmania infantum depends on tightly regulated signaling pathways to sustain growth, stress adaptation, and infectivity. Glycogen synthase kinase 3 (GSK-3) is a conserved serine/threonine kinase involved in multiple cellular processes; however, its functional role in Leishmania remains poorly defined. Here, we investigated the role of the short isoform of GSK-3 (GSK-3s) in L. infantum using a CRISPR/Cas9-based genetic approach. Attempts to generate chromosomal GSK-3s null mutants were unsuccessful, suggesting that GSK-3s is essential for parasite viability. Chromosomal gene deletion was achieved only after episomal complementation with L. braziliensis GSK-3s, generating a GSK3s-deficient knockdown L. infantum line (LiΔchrGSK3::LbGSK3). Quantitative PCR revealed a 3.8-fold and 9.6-fold reduction in GSK-3s transcript levels in this L. infantum line compared with Li::Cas9 and GSK3-overexpressing (Li::LbGSK3) parasites, respectively. Reduced GSK-3s expression in this parasite line resulted in severe growth impairment, morphological alterations, and cell-cycle dysregulation. GSK-3s knockdown promastigotes exhibited a 5.3-fold increase in resistance to trivalent antimony and a 1.77-fold increase in resistance to miltefosine, as well as a striking 111-fold increase in resistance to the GSK-3 inhibitor 6-bromo-5-methylindirubin-3'-oxime (6-BIO), supporting the specificity of this compound toward parasite GSK-3s. In addition, GSK-3s-deficient parasites displayed a 7.5-fold increase in tolerance to hydrogen peroxide-induced oxidative stress, accompanied by differential modulation of antioxidant defense gene transcripts. In infection models, GSK-3s knockdown parasites displayed reduced early infectivity in THP-1-derived macrophages and significantly lower splenic parasite burdens, with reductions of 1.5-fold in BALB/c mice and 2.5-fold in IFN-γ⁻/⁻ C57BL/6 mice. Collectively, these findings suggest that GSK-3s may play an important role in regulating L. infantum growth, stress adaptation, drug response, and infectivity, indicating its potential as a target for anti-leishmanial drug development.
PMID:
42405965
Bibliographic data and abstract were imported from PubMed on 06 Jul 2026.
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