In vivo profile of the human leukocyte microRNA response to endotoxemia

doi:10.1016/j.bbrc.2008.12.190

Biochemical and Biophysical Research Communications

Volume 380, Issue 3, 13 March 2009, Pages 437-441

https://doi.org/10.1016/j.bbrc.2008.12.190 Get rights and content

Abstract

To gain insight into microRNAs (miRNAs) involved in the regulation of the human innate immune response, we screened for differentially expressed miRNAs in circulating leukocytes in an in vivo model of acute inflammation triggered by Escherichia coli lipopolysaccharide (LPS) infusion.

Leukocyte RNA was isolated from venous blood samples obtained from healthy male volunteers before and 4 h after LPS-infusion. After fluorescence labeling, RNA samples were hybridized to microarrays containing capture probes for measuring the abundance of more than 600 human miRNAs. Target genes were predicted for differentially expressed miRNAs and then compared to changes in genome-wide expression levels, which had been established in a previous study.

Data analysis revealed that five miRNAs consistently responded to LPS-infusion, four of which were down-regulated (miR-146b, miR-150, miR-342, and let-7g) and one was up-regulated (miR-143). The miR-150 and mir-342 response was confirmed by real-time PCR. By correlating to measured LPS-induced changes of the leukocyte transcriptome, we next searched for predicted target genes, whose stability might be under (co-) control by these miRNAs. We found that the rapid transcriptional activation during acute inflammation of select genes, such as the gene encoding interleukin-1 receptor-associated kinase 2 (IRAK2) might be facilitated by decreased levels of LPS-responsive miRNAs. The increased level of miR-143 might be associated with the pronounced down-regulation of the B-cell CLL/lymphoma 2 (BCL2) gene expression during LPS endotoxemia, and could further be involved in the translational silencing of several other predicted inflammation-related target genes.

This is the first in vivo study to demonstrate relative abundance of miRNA levels in peripheral blood leukocytes during acute LPS-induced inflammation. The miRNAs and their potential target genes identified herein contribute to the understanding of the complex transcriptional program of innate immunity initiated by pathogens.

Section snippets

Materials and methods

Subjects and study design. The study population included three healthy, normal weight, drug free, male volunteers. Health was defined as the absence of a detectable disease as defined by normal results from history, physical examination, ECG, blood pressure, clinical chemistry, complete blood count, hepatitis virus serology, and HIV test. The study was approved by the Ethics Committee of the Medical University of Vienna, performed in accordance with the Declaration of Helsinki, the Austrian

LPS-induced differential miRNA levels

In the first step, we analyzed the microarray data by comparing RNA samples obtained from blood before (baseline) and 4 h after LPS-infusion. When employing paired t-test based statistics to discover differentially expressed miRNAs, we found seven miRNAs with different abundance after LPS-infusion compared with baseline (Fig. 1). Of these, four were detected at lower level after LPS (miR-146b, miR-150, miR-342, and let-7g), while three miRNAs (miR-143, and two proprietary miRNAs that are not in

Conclusions

In this study, we screened for changes induced by LPS to the in vivo miRNA profile of human circulating leukocytes and found that a relatively small number of miRNAs are subject to differential expression. At least for select targets, we were able to correlate miRNA changes to differential expression during LPS-induced inflammation. While it is not possible to draw definite conclusions as suggested targets of LPS-responsive miRNAs have not been validated experimentally, the predicted targets

References (33)

K. Takeda et al.
TLR signaling pathways
Semin Immunol
(2004)
G. Meister
MiRNAs get an early start on translational silencing
Cell
(2007)
E. Sonkoly et al.
MicroRNAs and immunity: novel players in the regulation of normal immune function and inflammation
Semin Cancer Biol
(2008)
F.B. Mayr et al.
Coagulation interventions in experimental human endotoxemia
Transl Res
(2006)
B.P. Lewis et al.
Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets
Cell
(2005)
B.P. Lewis et al.
Prediction of mammalian microRNA targets
Cell
(2003)
P. Maziere et al.
Prediction of microRNA targets
Drug Discov Today
(2007)
R. Medzhitov
Toll-like receptors and innate immunity
Nat Rev Immunol
(2001)
R. Medzhitov
Recognition of microorganisms and activation of the immune response
Nature
(2007)
K. Takeda et al.
Toll-like receptors
Annu Rev Immunol
(2003)

B. Beutler et al.

Innate immune sensing and its roots: the story of endotoxin

Nat Rev Immunol

(2003)

S.E. Calvano et al.

A network-based analysis of systemic inflammation in humans

Nature

(2005)

U. Prabhakar et al.

Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects

DNA Cell Biol

(2005)

W.M. Schmidt et al.

In-vivo effects of simvastatin and rosuvastatin on global gene expression in peripheral blood leucocytes in a human inflammation model

Pharmacogenet Genomics

(2008)

S. Talwar et al.

Gene expression profiles of peripheral blood leukocytes after endotoxin challenge in humans

Physiol Genomics

(2006)

M.P. Gantier et al.

Fine-tuning of the innate immune response by microRNAs

Immunol Cell Biol

(2007)

Cited by (106)

Trichoderma stromaticum spores induce autophagy and downregulate inflammatory mediators in human peripheral blood-derived macrophages
2022, Current Research in Microbial Sciences
Citation Excerpt :
MiR-146 and miR-155 are known by their ability to differentially contribute to endotoxin-inflammatory responses (Das Gupta et al., 2014). The first maintains a sub-inflammatory immune response or decrease the human leukocyte activities, while the second has pro-inflammatory characteristics (Nahid et al., 2009; Schmidt et al., 2009). Since miR-155 has an inflammatory effect while miR-146 has an anti-inflammatory effect (Testa et al., 2017; Zhao et al., 2016).
Trichoderma spp. are usually considered safe and normally used as biocontrol and biofertilization. Safety for human health is evaluated by several tests that detect various effects such as allergenicity, toxicity, infectivity, and pathogenicity. However, they do not evaluate the effects of the agent upon the immune system. The aim of this study was to investigate the interaction between T. stromaticum spores and mammalian cells to assess the immunomodulatory potential of the spores of this fungus. First, mouse macrophage cell line J774 and human macrophages were exposed to fungal spores and analyzed for structural features, through scanning and transmission electron microscopy. Then, various analysis were performed in human macrophages as to their effect in some functional and molecular aspects of the immune system through immunocytochemistry, flow cytometry and gene expression assays. We demonstrated that T. stromaticum spores induces autophagy and autophagy-related genes (ATGs) and downmodulate inflammatory mediators, including ROS, NLRP3, the cytokines IL-1β, IL-18, IL-12 and IL-10, as well as TLR2, TLR4, miR-146b and miR-155, which may lead to an augmented susceptibility to pathogens. Our study shows the extension of damages the biofungicide Tricovab® can cause in the innate immune response. Further studies are necessary to elucidate other innate and adaptive immune responses and, consequently, the safety of this fungus when in contact with humans.
MicroRNA-150 inhibits myeloid-derived suppressor cells proliferation and function through negative regulation of ARG-1 in sepsis
2021, Life Sciences
Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection. The majority of sepsis-related deaths occur during late sepsis, which presents as a state of immunosuppression. Myeloid-derived suppressor cells (MDSCs) have been reported to promote immunosuppression during sepsis. Here we aim to understand the role of microRNAs in regulating MDSCs proliferation and immunosuppression function during sepsis.
Murine sepsis model was established using cecal ligation and puncture (CLP). A microarray was used to identify microRNAs with differential expression in murine sepsis. The effect of microRNA-150 on MDSCs proliferation and function was then evaluated. 140 multiple trauma patients from Tongji Hospital and 10 healthy controls were recruited. Peripheral blood samples were taken and the serum level of miR-150 was measured.
In the murine model of sepsis, MDSCs expansion was noted in the spleen and bone marrow, while expression of miR-150 in MDSCs decreased. Replenishing miR-150 inhibited the expansion of MDSCs in both monocytic and polymorphonuclear subpopulations, as well as decreasing the immunosuppressive function of MDSCs, through down-regulation of ARG1. Both pro-inflammatory cytokine IL-6 and anti-inflammatory cytokines TGF-β and IL-10 were reduced by miR-150. In human, the serum level of miR-150 was down-regulated in septic patients and elevated in non-septic trauma patients compared to healthy controls.
Our study showed that MiR-150 is down-regulated during sepsis. Replenishing miR-150 reduces the immunosuppression function of MDSCs by down-regulating ARG1 in late sepsis. MiR-150 might serve as a potential therapeutic option for sepsis.
Circulating plasma microRNAs dysregulation and metabolic endotoxemia induced by a high-fat high-saturated diet
2020, Clinical Nutrition
High-fat diet increase two to three times the plasma lipopolysaccharide (LPS) levels and induce subclinical inflammation. Diet can modify gene expression due to epigenetic processes related to MicroRNAs (miRNAs). MicroRNAs (miRNAs) play important role in the post-transcriptional mechanisms involved in regulation of expression of genes related to the inflammatory response. Also, diet can indirectly induce post-transcriptional regulation of gene expression by miRNAs, which may affect the risk for the development of chronic diseases.
This study investigated the effect of high-fat high-saturated meal ingestion on plasma miRNA expression and LPS levels during the postprandial period in healthy women.
An interventional study was carried out in which a high-fat breakfast (1067.45 kcal), composed mainly of saturated fatty acids (56 g), and 500 mL of water, was offered. Blood samples were collected at baseline and 1, 3 and 5 h after meal intake. The studied population consisted of healthy women (n = 11), aged between 20 and 40 years, and body mass index (BMI) between 18.5 and 25 kg/m². Plasma levels of lipid profile, cytokines, adhesion molecules, and LPS were measured at the 3 time points. A profile of 752 human plasma miRNA expression was analyzed by real-time PCR assay. These analyzes were performed for all blood collection time-points.
Expression profile analysis revealed 33 differentially expressed plasma circulating miRNAs compared to that of the control group. MiR-145-5p and miR-200 were differentially modulated in all time-points post meal consumption. In addition, there was a significant increase in plasma LPS, triglycerides, myristic and palmitic saturated fatty acids levels at the 3 time-points in comparison with the control basal levels. We also observed increased levels of the plasma tumor necrosis factor alpha (TNF-α) cytokine and the vascular cell adhesion molecule 1 (VCAM-1) levels after 5 h post meal ingestion.
Ingestion of high-fat high-saturated meal was able to induce metabolic endotoxemia and increase the expression of pro-inflammatory molecules such as TNF-alpha and VCAM-1, as well as modulating circulating miRNAs possibly controlling inflammatory and lipid metabolism proteins at the postprandial period.
MiR-150 attenuates LPS-induced acute lung injury via targeting AKT3
2019, International Immunopharmacology
Citation Excerpt :
Our results inferred that miR-150 inhibited JNK and NF-κB pathways probably through targeting AKT3 pathway. In recent years, miR-150 has been identified to be implicated in host response to LPS [9]. Here, we firstly demonstrated that miR-150 was down-regulated, and inversely correlated with the disease severity and 28-day survival in patients with ARDS.
Acute lung injury (ALI) and its severe manifestation of acute respiratory distress syndrome (ARDS) in human lung are induced by inflammatory cytokines and endogenous factors such as miRNAs. However, the role of miR-150 in lipopolysaccharide (LPS)-induced ALI is not clear. Here, we found miR-150 expression was significantly reduced in the serum of patients with ARDS, and negatively associated with the disease severity and 28-day survival of ARDS. In vivo, miR-150 decreased total cell and neutrophil counts, and production of inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) as well as levels of total protein, albumin and IgM in the bronchoalveolar lavage (BAL) fluid in LPS-induced ALI mice. Meanwhile, miR-150 improved the 72 h survival rate of LPS-induced ALI mice. In-vitro assays demonstrated that miR-150 alleviated LPS-induced A549 cell apoptosis, autophagy, and release of inflammatory cytokines. Further, AKT3 was a direct target of miR-150. Silencing of AKT3 partially reversed LPS-induced A549 cell injury, and enhanced the protective effects of miR-150. In addition, miR-150 or si-AKT3 effectively inhibited the phosphorylation levels of c-Jun N-terminal kinase (JNK) and nuclear factor-κB (NF-κB) (p65 and IκBα). In conclusion, miR-150 alleviated LPS-induced acute lung injury via directly targeting AKT3 expression or regulating JNK and NF-κB pathways, which may be a promising therapeutic strategy to treat ALI/ARDS.
Integrative mRNA-miRNA interaction analysis associated with the immune response of Epinephelus coioddes to Vibrio alginolyticus infection
2019, Fish and Shellfish Immunology
MicroRNAs (miRNAs) are a kind of small non-coding RNAs that have been reported to play a vital role in mediating host-pathogen interactions. High-throughput sequencing technology was applied to identify and illuminate mRNAs and miRNAs from grouper infected with Vibrio alginolyticus. The KEGG pathway enrichment analysis showed that the most significate DEGs are associated with Toll-like receptor signaling pathway and NOD-like receptor signaling pathway. We obtained 374 known miRNAs and 116 novel miRNAs. During them, there are 31 up-regulated miRNAs and 93 down-regulated miRNAs. miRNA-mRNA GO and KEGG analysis show that there are 90 miRNAs associated with the immune system. The target genes of immune-related miRNAs (miR-142, miR-146, miR-150, miR-155, miR-203, miR-205, miR-24, miR-31) and genes (CD80, IL-2, AMPK, PI3K) in Epinephelus coioddes were predicted and validated. This study provides an opportunity to further understanding the molecular mechanisms especially the immune system of miRNA regulation in Epinephelus coioddes host-pathogen interactions.
Angiopoietin-like 8 (ANGPTL8) expression is regulated by miR-143-3p in human hepatocytes
2019, Gene
Citation Excerpt :
Previous studies have shown that miR-143 levels are higher in vascular smooth muscles cells (SMCs) from diabetic individuals compared to healthy controls, but in vitro treatment of SMCs with high glucose or insulin produced no significant effect on miRNA expression (Riches et al., 2014). Similarly, miR-143-3p upregulation in the presence of LPS in the current study is consistent with findings of increased circulating leukocyte levels of the miRNA in healthy male volunteers following LPS infusion (Schmidt et al., 2009). In that study, miR-143 levels were also associated with downregulation of pro-inflammatory genes.
Angiopoietin-like protein 8 (ANGPTL8) is associated with reduced HDL-cholesterol levels and may contribute to the development of dyslipidemia. Factors regulating ANGPTL8 expression remain poorly understood. Here we analyzed the relationship between miRNA-143-3p and ANGPTL8 in liver cells. Using target prediction algorithms, we identified a putative binding site for miR-143-3p in the ANGPTL8 3′ untranslated region (3’UTR). Exogenous miR-143-3p interacted with the ANGPTL8 3’UTR to downregulate its expression compared to scrambled sequence control. Transfection of HepG2 cells with miR-143-3p mimic or siRNA resulted in decreased or increased ANGPTL8 transcript and protein levels, respectively. Treatment of HepG2 cells with 30 mM glucose, 100 nM insulin, or 75 ng/ml lipopolysaccharide to mimic hyperglycemic, hyperinsulinemic, and proinflammatory conditions corresponded with increased miR-143-3p and ANGPTL8 levels. Inhibition of miR-143-3p amplified ANGPTL8 response to these treatments, suggesting that the miRNA acts to suppress ANGPTL8 expression under metabolically distorted conditions. These results, combined with growing evidence supporting a role for ANGPTL8 in the regulation of HDL-C metabolism, provide a better understanding of the molecular mechanisms underlying ANGPTL8 expression.

View all citing articles on Scopus

View full text

In vivo profile of the human leukocyte microRNA response to endotoxemia

Abstract

Section snippets

Materials and methods

LPS-induced differential miRNA levels

Conclusions

Semin Immunol

Cell

Semin Cancer Biol

Transl Res

Cell

Cell

Drug Discov Today

Toll-like receptors and innate immunity

Nat Rev Immunol

Recognition of microorganisms and activation of the immune response

Nature

Toll-like receptors

Annu Rev Immunol

Innate immune sensing and its roots: the story of endotoxin

Nat Rev Immunol

A network-based analysis of systemic inflammation in humans

Nature

Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects

DNA Cell Biol

In-vivo effects of simvastatin and rosuvastatin on global gene expression in peripheral blood leucocytes in a human inflammation model

Pharmacogenet Genomics

Gene expression profiles of peripheral blood leukocytes after endotoxin challenge in humans

Physiol Genomics

Fine-tuning of the innate immune response by microRNAs

Immunol Cell Biol