Dados do Trabalho

Título

HIF-1α interaction with HIF-1Α antisense long non-coding RNA shift response in macrophages during Leishmania infantum infection

Objetivo (s)

In this work, we investigated the correlation of hypoxia inducible factor 1 alpha (HIF1A) and HIF-1α antisense long non-coding RNAs (HIF1A-AS2, HIF1A-AS3) gene expression in macrophages infected with Leishmania infantum or stimulated with TLRs agonists.

Material e Métodos

We analyzed the HIF1A, HIF1A-AS2, HIF1A-AS3 gene expression in human THP-1-derived macrophages infected with L. infantum NCL (MOI 5:1) or stimulated with LPS (100ng/mL), poly(I:C) (100ng/mL) or gardiquimod (500ng/mL) for 4 and 24 h by RT-qPCR. We evaluated the HIF-1α expression in macrophages infected with L. infantum or TLRs agonists by flow cytometry. Next, we investigated the impact of HIF1A-AS3 knockdown and overexpression to infection and gene modulation using CRISPR-Cas9 strategy.

Resultados e Conclusão

In THP-1 macrophages infected with L. infantum, we observed reduced levels of HIF1A, HIF1A-AS2 and HIF1A-AS3 after 4 and 24h. Interestingly, LPS stimulation of macrophages increased the levels of HIF1A, HIF1A-AS2 and HIF1A-AS3 after 4 and 24h, but not with poly(I:C), and gardiquimod stimuli. Also, TLR3, TLR4 and TLR7 stimulation of THP-1 macrophages with poly(I:C), LPS and gardiquimod, respectively, modulated the genes hexokinase II(HKII), pyruvate dehydrogenase kinase 1(PDK1), pyruvate kinase M2 subtype(PKM2) and lactate dehydrogenase A(LDHA) expression.The L. infantum infection modulated these genes in a distinct way. The infection and LPS stimulation increased the frequency of HIF-1α+ cells. The knockdown and overexpression of HIF1A-AS3 regulate HIF1A and PDK1 expression.
Our initial data suggest that infection of human macrophages with L.infantum, reduces HIF1A, HIF1A-AS2 and HIF1A-AS3 expression, working in a distinct way of LPS. TLRs stimulation by poly(I:C), LPS and gardiquimod modulates HIF-1α target-genes from glycolysis and lncRNAs HIF1A-AS3.

Palavras-chave

leishmania, HIF-1α, glycolysis, lncRNAs

Agradecimentos

CAPES, FAPESP and CNPq