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

Área

Eixo 06 | Protozooses

Categoria

Concorrer ao Prêmio Jovem Pesquisador - Doutorado

Autores

Jonathan Miguel Zanatta, Juliane Cristina Ribeiro Fernandes, Stephanie Maia Acuña, Clara Andrade Teixeira, Carolina Manganeli Polonio, Tiago Francisco Da Silva, Lilian Gomes De Oliveira, Jean Pierre Schatzmann Peron, Sandra Marcia Muxel