Identification of suitable controls for miRNA quantification in T-cells and whole blood cells in sepsis

Hirschberger, Simon; Hübner, Max; Strauß, Gabriele; Effinger, David; Bauer, Michael; Weis, Sebastian; Giamarellos‐Bourboulis, Evangelos J.; Kreth, Simone

doi:10.1038/s41598-019-51782-w

Cited by 13 publications

(11 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although miR-150 has been previously revealed to serve key roles in diabetes and other autoimmune diseases ( 45 ), the role of this miRNA in sepsis remains unclear. In the present study, it was confirmed that miR-150 expression was downregulated in LPS-stimulated THP-1 cells, consistent with the reduced miR-150 expression observed in patients with sepsis ( 46 ). Results from the present study also demonstrated that miR-150 functioned as a negative regulator of the macrophage inflammatory response by suppressing the production of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α.…”

Section: Discussionsupporting

confidence: 91%

Anti‑inflammatory effects of miR‑150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment

et al. 2021

View full text Add to dashboard Cite

Sepsis is a condition that is associated with high rates of mortality. It is characterized by serious systemic inflammatory responses induced by pathogenic invasion. Although microRNA-150 (miR-150) has been previously reported to be involved in the modulation of sepsis, the underlying molecular mechanism in sepsis remains poorly understood. In the present study, the human monocytic cell line THP-1 was treated with LPS to mimic sepsis in vitro, following which miR-150 and STAT1 expression were measured using reverse transcription-quantitative PCR or western blotting. Secretion of inflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) into the medium were measured by ELISA. The potential relationship between STAT1 and miR-150 was determined using dual-luciferase reporter and RNA immunoprecipitation assays. miR-150 expression was found to be was downregulated by LPS treatment in THP-1 cells in both dose-and time-dependent manners. LPS treatment also induced IL-1β, IL-6 and TNF-α secretion in a manner that could be inhibited by miR-150 overexpression and enhanced by transfection with the miR-150 inhibitor. miR-150 was revealed to directly target STAT1 by negatively regulating its expression. In addition, STAT1 expression was demonstrated to be upregulated by LPS treatment. STAT1 overexpression reversed the inhibitory effects of miR-150 overexpression on IL-1β, IL-6 and TNF-α secretion whilst STAT1 knockdown attenuated IL-1β, IL-6 and TNF-α secretion induced by miR-150 inhibitor transfection. In conclusion, the present study suggested that miR-150 regulates the inflammatory response in macrophages following LPS challenge by regulating the expression of STAT1.

show abstract

Section: Discussionsupporting

confidence: 91%

Anti‑inflammatory effects of miR‑150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Expression of mRNA was determined using a LightCycler 480 instrument (Roche Diagnostics, Mannheim, Germany) as previously described (Möhnle et al , 2018 ; Hirschberger et al , 2019 ). All analyses were calculated using duplicate values.…”

Section: Methodsmentioning

confidence: 99%

Very‐low‐carbohydrate diet enhances human T‐cell immunity through immunometabolic reprogramming

et al. 2021

Self Cite

View full text Add to dashboard Cite

Very‐low‐carbohydrate diet triggers the endogenous production of ketone bodies as alternative energy substrates. There are as yet unproven assumptions that ketone bodies positively affect human immunity. We have investigated this topic in an in vitro model using primary human T cells and in an immuno‐nutritional intervention study enrolling healthy volunteers. We show that ketone bodies profoundly impact human T‐cell responses. CD4+, CD8+, and regulatory T‐cell capacity were markedly enhanced, and T memory cell formation was augmented. RNAseq and functional metabolic analyses revealed a fundamental immunometabolic reprogramming in response to ketones favoring mitochondrial oxidative metabolism. This confers superior respiratory reserve, cellular energy supply, and reactive oxygen species signaling. Our data suggest a very‐low‐carbohydrate diet as a clinical tool to improve human T‐cell immunity. Rethinking the value of nutrition and dietary interventions in modern medicine is required.

show abstract

“…U6 is one of the most commonly employed, although its use is a matter of debate; it is not properly a miRNA [ 28 , 30 ]. Some studies have indicated its unsuitability for miRNA normalization due to expression instability in various experimental conditions [ 31 , 32 , 33 ]. Conversely, in other models, U6 expression levels are very stable among the sample groups, making it a valid endogenous control [ 30 , 34 ].…”

Section: Discussionmentioning

confidence: 99%

miRNA Expression Profiling in Subcutaneous Adipose Tissue of Monozygotic Twins Discordant for HIV Infection: Validation of Differentially Expressed miRNA and Bioinformatic Analysis

Bresciani

Squillace

Orsini

et al. 2022

IJMS

View full text Add to dashboard Cite

Combined AntiRetroviral Treatments (cARTs) used for HIV infection may result in varied metabolic complications, which in some cases, may be related to patient genetic factors, particularly microRNAs. The use of monozygotic twins, differing only for HIV infection, presents a unique and powerful model for the controlled analysis of potential alterations of miRNAs regulation consequent to cART treatment. Profiling of 2578 mature miRNA in the subcutaneous (SC) adipose tissue and plasma of monozygotic twins was investigated by the GeneChip® miRNA 4.1 array. Real-time PCR and ddPCR experiments were performed in order to validate differentially expressed miRNAs. Target genes of deregulated miRNAs were predicted by the miRDB database (prediction score > 70) and enrichment analysis was carried out with g:Profiler. Processes in SC adipose tissue most greatly affected by miRNA up-regulation included (i) macromolecular metabolic processes, (ii) regulation of neurogenesis, and (iii) protein phosphorylation. Furthermore, KEGG analysis revealed miRNA up-regulation involvement in (i) insulin signaling pathways, (ii) neurotrophin signaling pathways, and (iii) pancreatic cancer. By contrast, miRNA up-regulation in plasma was involved in (i) melanoma, (ii) p53 signaling pathways, and (iii) focal adhesion. Our findings suggest a mechanism that may increase the predisposition of HIV+ patients to insulin resistance and cancer.

show abstract

Identification of suitable controls for miRNA quantification in T-cells and whole blood cells in sepsis

Cited by 13 publications

References 62 publications

Anti‑inflammatory effects of miR‑150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment

Anti‑inflammatory effects of miR‑150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment

Very‐low‐carbohydrate diet enhances human T‐cell immunity through immunometabolic reprogramming

miRNA Expression Profiling in Subcutaneous Adipose Tissue of Monozygotic Twins Discordant for HIV Infection: Validation of Differentially Expressed miRNA and Bioinformatic Analysis

Contact Info

Product

Resources

About