miR-148a regulation interferes in inflammatory cytokine and parasitic load in canine leishmaniasis

Abstract

Canine leishmaniasis (CanL) is a severe public health threat. Infected animals mediate transmission of the Leishmania protozoan to humans via the sandfly’s bite during a blood meal. CanL progression depends on the degree of suppression of the immune response, possibly associated with microRNAs (miR), which can modulate mRNA translation into proteins and (consequently) regulate cell function. Increased miR-148a in splenic leukocytes (SL) of dogs with CanL was observed in previous studies, and in silico analysis, identified possible pathways involved in immune response regulation that are affected by this miR. Therefore, we evaluated the involvement of miR-148a in the regulation of TNF-α, IL-6, IL-12, IL-1β, iNOS, MHCII, CD80, CD3, T-bet, and GATA-3 transcription factors and their relationship with parasite load in SL of dogs with CanL. Splenic leukocytes obtained from healthy and diseased dogs were transfected with miR-148a mimic and inhibitor oligonucleotides. After 48 hours, expression levels of MHCII, CD80, iNOS, CD3, T-bet, and GATA-3 were evaluated by flow cytometry, and concentrations of TNF-α, IL-12, IL-6, and IL-1β were measured in culture supernatants by capture enzyme-linked immunosorbent assays. Transfection of SL with miR-148a mimics decreased iNOS levels in cells and TNF-α, IL-6, and IL-12 in the supernatants of cultured SL from CanL dogs. Interestingly, transfection with miR-148a inhibitor decreased parasite load in SL cells. These results suggest a direct or not regulatory role of this miR in the immune response to Leishmania infantum infection. We conclude that miR-148a can modulate immune responses by regulating inflammatory cytokines during CanL. Our results contribute to understanding the complex host/parasite interaction in CanL and could assist the development of treatments.

MicroRNA-21 and microRNA-148a affects PTEN, NO and ROS in canine leishmaniasis

“…We observed SLs transfected with the miR-148a inhibitor there was more significant NO production, in contrast miR-148a mimic also decrease iNOS in SL in CanL ( Rebech et al, 2023 ), and similar study observed that overexpression of miR-148a in peripheral blood cells of rats was associated with low iNOS ( Yin et al, 2021 ). NO production by macrophages participates in the elimination of L. infantum and can be involved in Th1-type immune responses ( Reza et al, 2019 ).…”

“…MiR-21 and miR-148a expression was evaluated previously in SL, and there was an increase in the expression of miR-21 ( Bragato et al, 2022 ) and miR-148a ( Rebech et al, 2023 ). SLs were cultured at 1.6 × 10 5 in 24-well plates for 48 h at 37 °C in 5% CO 2 .…”

“…Frontiers in Genetics frontiersin.org (Bragato et al, 2022) and miR-148a (Rebech et al, 2023). SLs were cultured at 1.6 × 10 5 in 24-well plates for 48 h at 37 °C in 5% CO 2 .…”

“…These findings suggest that L. infantum modulates the immune response via miR-21 expression as a survival mechanism ( Bragato et al, 2022 ). In addition, the decreased parasitic load in SL following the use of the miR-148a inhibitor suggests a critical role for this miR in developing microbicidal activity in CanL, and miR148a regulates immune response as demonstrated by reducing inducible nitric oxide synthase (iNOS), TNF-α, IL-6, and IL-12 ( Rebech et al, 2023 ).…”

Extracellular vesicle biogenesis of three‐dimensional human pluripotent stem cells in a novel Vertical‐Wheel bioreactor

“…Small RNA‐seq also identified miR‐21 in hiPSC‐EVs, which has a variety of functions including regulation of mechanotransduction, neuroprotection, neurite outgrowth and modulation of proinflammatory microglia (Deng et al., 2022 ; Hurwitz et al., 2018 ; Vichai & Kirtikara, 2006 ; Xia et al., 2020 ). For other identified miRNAs in hiPSC‐EVs, miR‐148a can modulate immune responses by regulating inflammatory cytokines (Rebech et al., 2023 ). miR‐20a may play anti‐inflammatory and anti‐apoptotic roles via deactivation of cytokine CXCL12 and CXC chemokine receptor (CXCR)4, NF‐κB (nuclear factor kappa light chain enhancer of activated B cells) signalling and extracellular signal‐regulated kinase (ERK)1/2 signalling (Zhang et al., 2020 ).…”