TGF-beta/atRA induced Tregs express a selected set of microRNAs involved in the repression of transcripts related to Th17 differentiation
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE93858 | GEO | 2017/01/20
SECONDARY ACCESSION(S): PRJNA362600
REPOSITORIES: GEO
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Project description:Regulatory T cells (Tregs) are essential regulators of immune tolerance. atRA and TGF-β can inhibit the polarization of naïve T cells into inflammatory Th17 cells, favoring the generation of stable iTregs, however the regulatory mechanisms involved are not fully understood. In this context, the roles of individual microRNAs in Tregs are largely unexplored. Naïve T cells were immunomagnetically isolated from umbilical cord blood and activated with anti-human CD2/CD3/CD28 beads in the presence of IL-2 alone (CD4Med) or with the addition of TGF-β and atRA (CD4TGF/atRA). As compared to CD4Med, the CD4TGF/atRA condition allowed the generation of highly suppressive CD4+CD25hiCD127-FOXP3hi iTregs. Microarray profiling allowed the identification of a set of microRNAs that are exclusively expressed upon TGF-β/atRA treatment and that are predicted to target a set of transcripts concordantly downregulated. This set of predicted targets were enriched for central components of IL-6/JAK/STAT and AKT-mTOR signaling, whose inhibition is known to play important roles in the generation and function of regulatory lymphocytes. Finally, we show that mimics of exclusively expressed miRs (namely miR-1299 and miR-30a-5p) can reduce the levels of its target transcripts, IL6R and IL6ST (GP130), and increase the percentage of FoxP3+ cells among CD4+CD25+/hi cells.
| S-EPMC5472579 | biostudies-literature
Project description:Regulatory T cells (Tregs) are essential regulators of immune tolerance. In light of its potential therapeutic use, in conditions involving transplant rejection and autoimmune disorders, understanding the mechanisms involved in the polarization between inflammatory and regulatory cells is of great importance. Although several studies have shown that atRA and TGF-β can inhibit the polarization of naïve T cells into inflammatory Th17 cells, favoring the generation of stable FOXP3 expressing iTregs, the exact regulatory mechanisms involved are not fully understood. Increasing evidences have suggested several roles for microRNAs in the development and function of different types of T helper cells; however, the roles of individual microRNAs in Tregs function and development are largely unexplored. Here, we explored how atRA may post-transcriptionally regulate distinct signaling pathways, through the induction of specific microRNAs, controlling the balance between Th17 and T regulatory cells. For this, CD4+CD25-CD45RA+ naïve T cells were immunomagnetically isolated from umbilical cord blood and activated with anti-human CD2/CD3/CD28 beads in the presence of IL-2 alone (CD4Med) or with the addition of TGF-β and atRA (CD4TGF/atRA). Immunophenotyping by flow cytometry, whole genome mRNAs and microRNAs profiling and functional characterizations were performed. We report the generation of highly suppressive CD4+CD25hiCD127-FOXP3+ iTregs in the CD4TGF/atRA condition, that exclusively express a set of microRNAs predicted to target important signaling pathways, including PI3K/Akt, mTOR, Toll-Like Receptor, and IL-6/Jak/Stat. Accordingly, percentages of IL-17 and STAT3 expressing cells were strikingly reduced in CD4TGF/atRA condition. Microarray and qPCR results showed that the expression of IL6R, IL6ST, JAK1 and STAT3 (heavily targeted by identified miRs) are all downregulated in CD4TGF/atRA, as compared to CD4med and naïve T cells. Finally, we show that selected synthetic mimics (namely, miR-23a, -30a, -636 and -1299) are able to knockdown IL6R and IL6ST transcript levels, functionally inhibiting IL-17 expression and favoring the generation of FOXP3+ cells. Our work adds novel evidences supporting the central roles played by microRNAs in the post-transcriptional regulation of major pathways involved in the generation and function of regulatory T cells.
2017-01-20 | GSE93857 | GEO
Project description:Regulatory T cells (Tregs) are essential regulators of immune tolerance. In light of its potential therapeutic use, in conditions involving transplant rejection and autoimmune disorders, understanding the mechanisms involved in the polarization between inflammatory and regulatory cells is of great importance. Although several studies have shown that atRA and TGF-β can inhibit the polarization of naïve T cells into inflammatory Th17 cells, favoring the generation of stable FOXP3 expressing iTregs, the exact regulatory mechanisms involved are not fully understood. Increasing evidences have suggested several roles for microRNAs in the development and function of different types of T helper cells; however, the roles of individual microRNAs in Tregs function and development are largely unexplored. Here, we explored how atRA may post-transcriptionally regulate distinct signaling pathways, through the induction of specific microRNAs, controlling the balance between Th17 and T regulatory cells. For this, CD4+CD25-CD45RA+ naïve T cells were immunomagnetically isolated from umbilical cord blood and activated with anti-human CD2/CD3/CD28 beads in the presence of IL-2 alone (CD4Med) or with the addition of TGF-β and atRA (CD4TGF/atRA). Immunophenotyping by flow cytometry, whole genome mRNAs and microRNAs profiling and functional characterizations were performed. We report the generation of highly suppressive CD4+CD25hiCD127-FOXP3+ iTregs in the CD4TGF/atRA condition, that exclusively express a set of microRNAs predicted to target important signaling pathways, including PI3K/Akt, mTOR, Toll-Like Receptor, and IL-6/Jak/Stat. Accordingly, percentages of IL-17 and STAT3 expressing cells were strikingly reduced in CD4TGF/atRA condition. Microarray and qPCR results showed that the expression of IL6R, IL6ST, JAK1 and STAT3 (heavily targeted by identified miRs) are all downregulated in CD4TGF/atRA, as compared to CD4med and naïve T cells. Finally, we show that selected synthetic mimics (namely, miR-23a, -30a, -636 and -1299) are able to knockdown IL6R and IL6ST transcript levels, functionally inhibiting IL-17 expression and favoring the generation of FOXP3+ cells. Our work adds novel evidences supporting the central roles played by microRNAs in the post-transcriptional regulation of major pathways involved in the generation and function of regulatory T cells.
2017-01-20 | GSE93851 | GEO
Project description:TGF-beta/atRA induced Tregs express a selected set of microRNAs involved in the repression of transcripts related to Th17 differentiation
| PRJNA362600 | ENA
Project description:Background50% of leprosy patients suffer from episodes of Type 1/ reversal reactions (RR) and Type 2/ Erythema Nodosum Leprosum (ENL) reactions which lead to morbidity and nerve damage. CD4+ subsets of Th17 cells and CD25+FOXP3+ regulatory T cells (Tregs) have been shown to play a major role in disease associated immunopathology and in stable leprosy as reported by us and others. The aim of our study was to analyze their role in leprosy reactions.Methodology and principle findingsQuantitative reverse transcribed PCR (qPCR), flowcytometry and ELISA were used to respectively investigate gene expression, cell phenotypes and supernatant levels of cytokines in antigen stimulated PBMC cultures in patients with stable disease and those undergoing leprosy reactions. Both types of reactions are associated with significant increase of Th17 cells and associated cytokines IL-17A, IL-17F, IL-21, IL-23 and chemokines CCL20, CCL22 as compared to matching stable forms of leprosy. Concurrently patients in reactions show reduction in FOXP3+ Treg cells as well as reduction in TGF-β and increase in IL-6. Moreover, expression of many T cell markers, cytokines, chemokines and signaling factors were observed to be increased in RR as compared to ENL reaction patients.ConclusionsPatients with leprosy reactions show an imbalance in Th17 and Treg populations. The reduction in Treg suppressor activity is associated withhigherTh17cell activity. The combined effect of reduced TGF-β and enhanced IL-6, IL-21 cytokines influence the balance between Th17 or Treg cells in leprosy reactions as reported in the murine models and autoimmune diseases. The increase in Th17 cell associated cytokines may contribute to lesional inflammation.
| S-EPMC4818038 | biostudies-literature
Project description:The hallmark of pancreatic ductal adenocarcinoma (PDAC) is abundant desmoplasia, which is orchestrated by pancreatic stellate cells (PSCs) and accounts for the majority of the stroma surrounding the tumour. Healthy PSCs are quiescent, but upon activation during disease progression, they adopt a myofibroblast-contractile phenotype and secrete and concomitantly reorganise the stiff extracellular matrix (ECM). Transforming growth factor ? (TGF-?) is a potent activator of PSCs, and its activation requires spatiotemporal organisation of cellular and extracellular cues to liberate it from an inactive complex with latent TGF-? binding protein (LTBP). Here we study the mechanical activation of TGF-? by PSCs in vitro by investigating LTBP-1 organisation with fibrillar fibronectin and show that all trans-retinoic acid (ATRA), which induces PSC quiescence, down-regulates the ability of PSCs to mechanically organise LTBP-1 and activate TGF-? through a mechanism involving myosin II dependent contractility. Therefore, ATRA inhibits the ability of PSCs to mechanically release active TGF-?, which might otherwise act in an autocrine manner to sustain PSCs in an active state and a tumour-favouring stiff microenvironment.
| S-EPMC4931506 | biostudies-literature
Project description:The capacity of IL-10 and Tregs in the inflammatory tumor microenvironment to impair anticancer Th1 immunity makes them attractive targets for cancer immunotherapy. IL-10 and Tregs also suppress Th17 activity, which is associated with poor prognosis in several cancers. However, previous studies have overlooked their potential contribution to the regulation of pathogenic cancer-associated inflammation. In this study, we investigated the origin and function of IL-10–producing cells in the tumor microenvironment using transplantable tumor models in mice. The majority of tumor-associated IL-10 was produced by an activated Treg population. IL-10 production by Tregs was required to restrain Th17-type inflammation. Accumulation of activated IL-10+ Tregs in the tumor required type I IFN signaling but not inflammatory signaling pathways that depend on TLR adapter protein MyD88 or IL-12 family cytokines. IL-10 production limited Th17 cell numbers in both spleen and tumor. However, type I IFN was required to limit Th17 cells specifically in the tumor microenvironment, reflecting selective control of tumor-associated Tregs by type I IFN. Thus, the interplay of type I IFN, Tregs, and IL-10 is required to negatively regulate Th17 inflammation in the tumor microenvironment. Therapeutic interference of this network could therefore have the undesirable consequence of promoting Th17 inflammation and cancer growth.
| S-EPMC3809773 | biostudies-literature
Project description:Cancer cells display an altered distribution of DNA methylation relative to normal cells. Certain tumor suppressor gene promoters are hypermethylated and transcriptionally inactivated, whereas repetitive DNA is hypomethylated and transcriptionally active. Little is understood about how the abnormal DNA methylation patterns of cancer cells are established and maintained. Here, we identify over 20 DNMT3B transcripts from many cancer cell lines and primary acute leukemia cells that contain aberrant splicing at the 5' end of the gene, encoding truncated proteins lacking the C-terminal catalytic domain. Many of these aberrant transcripts retain intron sequences. Although the aberrant transcripts represent a minority of the DNMT3B transcripts present, Western blot analysis demonstrates truncated DNMT3B isoforms in the nuclear protein extracts of cancer cells. To test if expression of a truncated DNMT3B protein could alter the DNA methylation patterns within cells, we expressed DNMT3B7, the most frequently expressed aberrant transcript, in 293 cells. DNMT3B7-expressing 293 cells have altered gene expression as identified by microarray analysis. Some of these changes in gene expression correlate with altered DNA methylation of corresponding CpG islands. These results suggest that truncated DNMT3B proteins could play a role in the abnormal distribution of DNA methylation found in cancer cells.
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Project description:Interleukin-23 (IL-23) is required for inflammatory Th17 cell function in experimental autoimmune encephalomyelitis (EAE), and IL-23 blockade reduces the number of effector Th17 cells in the CNS. We report that pro-inflammatory Th17 cells express high integrin ?3 that is IL-23 dependent. Integrin ?3 was not upregulated on all activated T cells; rather, integrin ?3 was upregulated along with its functional partner integrin ?v on effector Th17 cells and "ex-Th17" cells, and ?v?3(hi) ROR?t(+) cells expanded during EAE. Integrin ?v?3 inhibitors ameliorated clinical signs of EAE, and integrin ?3 deficiency on CD4(+) T cells alone was sufficient to block EAE induction. Furthermore, integrin-?3-deficient Th17 cells, but not Th1 cells, were impaired in their ability to induce EAE. Integrin ?3(-/-) T cells induced smaller demyelinated lesions and showed reduced spread and accumulation within the CNS, corresponding with impaired extracellular-matrix-mediated migration. Hence, integrin ?3 is required for Th17 cell-mediated autoimmune CNS inflammation.
| S-EPMC5627357 | biostudies-literature
Project description:T helper (Th) 17 cells and CD4(+) CD25(+) regulatory T (Treg) cells are supposed to be critically involved in regulating autoimmune and inflammatory diseases. The aim of this study was to investigate the Th17/Treg pattern in rats with gunpowder smog-induced acute lung injury. Wistar rats were equally randomized to three groups: normal control group, ALI 6 h group (smoke inhalation for 6 h) and ALI 24 h group (smoke inhalation for 24 h). We observed changes in cell counting in bronchoalveolar lavage fluid (BALF), alveolar-capillary membrane permeability and lung tissue pathology. Moreover, rats in ALI 6 h and ALI 24 h group showed increased expression of Th17 cell and related cytokines (IL-17 A, IL-6, TGF-β and IL-23). Meanwhile, Treg prevalence and related cytokines (IL-10, IL-2 and IL-35) were decreased. Consequently, the ratio of Th17/Treg was higher after smoke inhalation. Additionally, Th1 cell decreased while Th2 cell increased at 6 h and 24 h after smoke inhalation. In conclusion, Th17/Treg imbalance exists in rats with smoke inhalation-induced acute lung injury, suggesting its potential role in the pathogenesis of this disease.
| S-EPMC4756332 | biostudies-literature