Enforced STAT3 and PU.1 Expression Activates Type I Interferon-Producing Cell and Dendritic Cell Differentiation Programs in Megakaryocyte/Erythrocyte Progenitors.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 201-201
Author(s):  
Nobuyuki Onai ◽  
Aya Onai ◽  
Roxane Tussiwand ◽  
Antonio Lanzavecchia ◽  
Markus G Manz
Keyword(s):  
Transcription Factors ◽  
Dendritic Cell ◽  
Expression Vectors ◽  
Cytokine Signaling ◽  
Type I ◽  
Mrna Levels ◽  
Developmental Potential ◽  
Cell Fate Decisions ◽  
Dendritic Cell Differentiation ◽  
Flt3 Expression

Abstract A standing question in early hematopoiesis is whether cytokine signaling is sufficient to induce cell fate decisions. Previously, we have reported that enforced expression of human Flt3 in Flt3 negative megakaryocyte/erythrocyte progenitors (MEPs) rescues their interferon producing cell (IPC) and dendritic cell (DC) developmental potential: Human Flt3-signaling in MEPs leads to up-regulation of IPC, DC, and myelomonocytic development affiliated genes such as, STAT3, PU.1, and G-CSFR/M-CSFR/GM-CSFR, and activates differentiation to these lineages. To test whether single Flt3 downstream activated genes would be sufficient to cause the same effects, we transduced murine STAT3 and PU.1 cDNA into MEPs using retrovirus expression vectors, respectively. Both STAT3- and PU.1-transduced MEPs were capable to differentiate into IPCs and DCs in SCF and TPO supplemented media, and, with even higher efficiency, in SCF, TPO, and Flt3L supplemented cultures. While human Flt3-transduced MEPs maintained megakaryocyte-erythrocyte developmental potential and gained IPC, DC, and myelomonocytic cell potential, STAT3- and PU.1-transduced MEPs lost both megakaryocyte and erythrocyte developmental options, indicating that strong signaling of these transcription factors fully converted MEPs to the IPC, DC, and myelomonocytic lineages. Consistently, GATA-1 expression was down-regulated, and C/EBPα mRNA was up-regulated in STAT3- and PU.1-transduced MEPs. Interestingly, STAT3- and PU.1 over-expression in MEPs led to up-regulation of Flt3 mRNA levels, suggesting a self-sustaining effect of Flt3 signaling-cascade induced Flt3 expression. Thus, enforced expression of STAT3 and PU.1 in MEPs reprogrammed them to differentiate into IPCs, DCs and myelomonocytic cell lineages and inhibit Meg/E-lineage potential. Based on these data, we propose a model where Flt3 positive progenitor cells that locate in Flt3L rich environments will be directed to develop into IPCs and DCs. This process might be enhanced by a self-sustaining mechanism where Flt3 downstream transcription factors as STAT3 and PU.1 in turn maintain Flt3 expression.

scholarly journals Rapid muscle atrophy response to unloading: pretranslational processes involving MHC and actin

2009 ◽  
Vol 107 (4) ◽  
pp. 1204-1212 ◽  
Author(s):  
Julia M. Giger ◽  
Paul W. Bodell ◽  
Ming Zeng ◽  
Kenneth M. Baldwin ◽  
Fadia Haddad
Keyword(s):  
Mrna Expression ◽  
Serum Response Factor ◽  
Striated Muscle ◽  
Weight Bearing ◽  
Hindlimb Suspension ◽  
Cytokine Signaling ◽  
Significant Shift ◽  
Type I ◽  
Mrna Levels

Skeletal muscles, especially weight-bearing muscles, are very sensitive to changes in loading state. The aim of this paper was to characterize the dynamic changes in the unloaded soleus muscle in vivo following a short bout of hindlimb suspension (HS), testing the hypothesis that transcriptional events respond early to the atrophic stimulus. In fact, we observed that after only 1 day of HS, primary transcript levels of skeletal α-actin and type I myosin heavy chain (MHC) genes were significantly reduced by more than 50% compared with ground control levels. The degree of the decline for the mRNA expression of actin and type I MHC lagged behind that of the pre-mRNA levels after 1 day of HS, but by 2 and 7 days of HS, large decreases were observed. Although the faster MHC isoforms, IIx and IIb, began to be expressed in soleus after 1 day of HS, a relatively significant shift in mRNA expression from the slow MHC isoform type I toward these fast MHC isoforms did not emerge until 7 days of HS. One day of HS was sufficient to show significant decreases in mRNA levels of putative signaling factors serum response factor (SRF), suppressor of cytokine signaling-3 (SOCS3), and striated muscle activator of Rho signaling (STARS), although transcription factors yin-yang-1 (YY1) and transcriptional enhancing factor-1 (TEF-1) were not significantly affected by HS. The protein levels of actin and type I MHC were significantly decreased after 2 days of HS, and SRF protein was significantly decreased after 7 days HS. Our results show that after only 1 day of unloading, pre-mRNA and mRNA expression of muscle proteins and muscle-specific signaling factors are significantly reduced, suggesting that the downregulation of the synthesis side of the protein balance equation that occurs in atrophying muscle is initiated rapidly.

IL-17 stimulates MMP-1 expression in primary human cardiac fibroblasts via p38 MAPK- and ERK1/2-dependent C/EBP-β, NF-κB, and AP-1 activation

2007 ◽  
Vol 293 (6) ◽  
pp. H3356-H3365 ◽  
Author(s):  
Dolores M. Cortez ◽  
Marc D. Feldman ◽  
Srinivas Mummidi ◽  
Anthony J. Valente ◽  
Bjorn Steffensen ◽  
...  
Keyword(s):  
Transcription Factor ◽  
P38 Mapk ◽  
Western Blotting ◽  
Cardiac Fibroblasts ◽  
Critical Role ◽  
Dominant Negative ◽  
Expression Vectors ◽  
Type I ◽  
Mrna Levels ◽  
Human Cardiac Fibroblasts

Matrix metalloproteinases (MMPs) degrade collagen and mediate tissue remodeling. The novel cytokine IL-17 is expressed during various inflammatory conditions and modulates MMP expression. We investigated the effect of IL-17 on MMP-1 expression in primary human cardiac fibroblasts (HCF) and delineated the signaling pathways involved. HCF were treated with recombinant human IL-17. MMP-1 expression was analyzed by Northern blotting, RT-quantitative PCR, Western blotting, and ELISA; transcriptional induction and transcription factor binding by EMSA, ELISA, and reporter assay; and p38 MAPK and ERK1/2 activation by protein kinase assays and Western blotting. Signal transduction pathways were investigated using pharmacological inhibitors, small interfering RNA (siRNA), and adenoviral dominant-negative expression vectors. IL-17 stimulated MMP-1 gene transcription, net mRNA levels, protein, and promoter-reporter activity in HCF. This response was blocked by IL-17 receptor-Fc chimera and IL-17 receptor antibodies, but not by IL-6, TNF-α, or IL-1β antibodies. IL-17-stimulated type I collagenase activity was inhibited by the MMP inhibitor GM-6001 and by siRNA-mediated MMP-1 knockdown. IL-17 stimulated activator protein-1 [AP-1 (c-Fos, c-Jun, and Fra-1)], NF-κB (p50 and p65), and CCAAT enhancer-binding protein (C/EBP)-β DNA binding and reporter gene activities, effects attenuated by antisense oligonucleotides, siRNA-mediated knockdown, or expression of dominant-negative signaling proteins. Inhibition of AP-1, NF-κB, or C/EBP activation attenuated IL-17-stimulated MMP-1 expression. IL-17 induced p38 MAPK and ERK1/2 activation, and inhibition by SB-203580 and PD-98059 blunted IL-17-mediated transcription factor activation and MMP-1 expression. Our data indicate that IL-17 induces MMP-1 in human cardiac fibroblasts directly via p38 MAPK- and ERK-dependent AP-1, NF-κB, and C/EBP-β activation and suggest that IL-17 may play a critical role in myocardial remodeling.

The Role of EHZF in Erythroid Differentiation of K562 Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1311-1311
Author(s):  
Etsuko Matsubara ◽  
Ikuya Sakai ◽  
Jun Yamanouchi ◽  
Taichi Azuma ◽  
Hiroshi Narumi ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Zinc Finger Protein ◽  
K562 Cells ◽  
Erythroid Differentiation ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Expression Vectors ◽  
Mrna Levels ◽  
Control Clone

Abstract Early hematopoietic zinc finger protein (EHZF), the human homolog to mouse Evi3, is a transcription factor with 30 zinc fingers and a highly conserved N-terminal FOG repression motif. The EHZF mRNA expression is abundant in early hematopoietic progenitors and declines during differentiation. In most of the leukemic cells from acute myelogenous leukemia patients, significant levels of EHZF mRNA are detected. These findings suggest that EHZF play an important role in hematopoietic differentiation. In the present study we investigated whether siRNA-mediated depletion of EHZF affected the erythroid differentiation of K562 cells. Three kind of EHZF siRNA were designed and their expression vectors were constructed. After transient expression of these siRNAs by electroporation, hemoglobinization of K562 cells were analyzed by Benzidine staining. In all of these siRNAs introduced cells, the levels of hemoglobinizations were abundant compared to control cells, which suggested that EHZF might influence the erythroid differentiation of K562 cells. To further analyze the effects of EHZF siRNA in erythroid differentiation, we established K562 clones in which EHZF was depleted by siRNA. EHZF siRNA expression vectors were introduced into K562 cells by electroporation and the expression of EHZF mRNA were examined by RT-PCR in G418 resistant clones. The depressions of EHZF mRNA in the clones were confirmed by Northern blotting. We isolated several EHZF depleted clones in each kind of EHZF siRNA introduced cells. Most of the EHZF depleted clones showed marked hemoglobinization by Benzidine staining compared to control clones, and surface expressions of glycophorin A were also increased in these clones. These results confirmed the relevant role of EHZF in erythroid differentiation of K562 cells. Comparison of mRNA expressions between an EHZF depleted clone and a control clone using DNA array showed increase expressions of hemoglobin alpha, beta, delta mRNA in EHZF depleted clone compared to control clone. Several transcription factors which are involved in erythroid differentiation also showed the difference mRNA levels between two clones. Studies are underway to elucidate the mechanisms underlying the role of EHZF in erythroid differentiation, the relationship to other transcription factors (GATA-1, GFi-1B, FOG-1 etc) which involved in erythroid differentiation.

S1PR4 is required for plasmacytoid dendritic cell differentiation

2015 ◽  
Vol 396 (6-7) ◽  
pp. 775-782 ◽  
Author(s):  
Christina Dillmann ◽  
Javier Mora ◽  
Catherine Olesch ◽  
Bernhard Brüne ◽  
Andreas Weigert
Keyword(s):  
Dendritic Cell ◽  
Cell Biology ◽  
Cell Function ◽  
Immune Cell ◽  
Plasmacytoid Dendritic Cell ◽  
Type I ◽  
Hematopoietic Stem ◽  
Dendritic Cell Differentiation ◽  
Sphingosine 1 Phosphate

Abstract The sphingolipid sphingosine-1-phosphate (S1P) has various functions in immune cell biology, regulating survival, proliferation, and, most prominently, migration. S1P couples to five G protein-coupled receptors (S1PR1–5) to transduce its effects on immune cell function. Expression of S1PR4 is restricted to immune cells. However, its impact on immune cell biology is largely elusive. In the current study, we intended to answer the question of whether S1P might affect plasmacytoid dendritic cell (pDC) migration, which dominantly express S1PR4. pDC are highly specialized cells producing large amounts of type I interferon in response to TLR7/9 ligands after viral infection or during autoimmunity. Surprisingly, we noticed a reduced abundance of pDC, particularly CD4- pDC, in all organs of S1PR4-deficient vs. wildtype mice. This effect was not caused by altered migration of mature pDC, but rather a reduced potential of pDC progenitors, especially common DC progenitors, to differentiate into pDCs. In vitro studies suggested that reduced S1PR4-deficient pDC progenitor differentiation into mature pDC might be explained by both migration and differentiation of pDC progenitors in the bone marrow. As S1PR4 also affected the differentiation of CD34+ human hematopoietic stem cells into pDC, interfering with S1PR4 might be useful to reduce pDC numbers during autoimmunity.

scholarly journals Pluripotency Factors on Their Lineage Move

2016 ◽  
Vol 2016 ◽  
pp. 1-16 ◽  
Author(s):  
Clair E. Weidgang ◽  
Thomas Seufferlein ◽  
Alexander Kleger ◽  
Martin Mueller
Keyword(s):  
Transcription Factors ◽  
Cell Fate ◽  
Ground State ◽  
Regulatory Networks ◽  
Embryonic Stem ◽  
Lineage Specification ◽  
Developmental Potential ◽  
Cell Fate Decisions ◽  
Complex Interactions ◽  
Great Knowledge

Pluripotent stem cells are characterised by continuous self-renewal while maintaining the potential to differentiate into cells of all three germ layers. Regulatory networks of maintaining pluripotency have been described in great detail and, similarly, there is great knowledge on key players that regulate their differentiation. Interestingly, pluripotency has various shades with distinct developmental potential, an observation that coined the term of a ground state of pluripotency. A precise interplay of signalling axes regulates ground state conditions and acts in concert with a combination of key transcription factors. The balance between these transcription factors greatly influences the integrity of the pluripotency network and latest research suggests that minute changes in their expression can strengthen but also collapse the network. Moreover, recent studies reveal different facets of these core factors in balancing a controlled and directed exit from pluripotency. Thereby, subsets of pluripotency-maintaining factors have been shown to adopt new roles during lineage specification and have been globally defined towards neuroectodermal and mesendodermal sets of embryonic stem cell genes. However, detailed underlying insights into how these transcription factors orchestrate cell fate decisions remain largely elusive. Our group and others unravelled complex interactions in the regulation of this controlled exit. Herein, we summarise recent findings and discuss the potential mechanisms involved.

scholarly journals Differential Activity of Type I Interferon Subtypes for Dendritic Cell Differentiation

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e58465 ◽  
Author(s):  
Geneviève Garcin ◽  
Yann Bordat ◽  
Paul Chuchana ◽  
Danièle Monneron ◽  
Helen K. W. Law ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Dendritic Cell ◽  
Type I Interferon ◽  
Type I ◽  
Dendritic Cell Differentiation

scholarly journals Transcriptome Analysis of Hypertrophic Heart Tissues from Murine Transverse Aortic Constriction and Human Aortic Stenosis Reveals Key Genes and Transcription Factors Involved in Cardiac Remodeling Induced by Mechanical Stress

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Peng Yu ◽  
Baoli Zhang ◽  
Ming Liu ◽  
Ying Yu ◽  
Ji Zhao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Aortic Stenosis ◽  
Mechanical Stress ◽  
Cardiac Remodeling ◽  
Interaction Network ◽  
Cytokine Signaling ◽  
Transverse Aortic Constriction ◽  
Aortic Constriction ◽  
Genomic Changes

Background. Mechanical stress-induced cardiac remodeling that results in heart failure is characterized by transcriptional reprogramming of gene expression. However, a systematic study of genomic changes involved in this process has not been performed to date. To investigate the genomic changes and underlying mechanism of cardiac remodeling, we collected and analyzed DNA microarray data for murine transverse aortic constriction (TAC) and human aortic stenosis (AS) from the Gene Expression Omnibus database and the European Bioinformatics Institute. Methods and Results. The differential expression genes (DEGs) across the datasets were merged. The Venn diagrams showed that the number of intersections for early and late cardiac remodeling was 74 and 16, respectively. Gene ontology and protein–protein interaction network analysis showed that metabolic changes, cell differentiation and growth, cell cycling, and collagen fibril organization accounted for a great portion of the DEGs in the TAC model, while in AS patients’ immune system signaling and cytokine signaling displayed the most significant changes. The intersections between the TAC model and AS patients were few. Nevertheless, the DEGs of the two species shared some common regulatory transcription factors (TFs), including SP1, CEBPB, PPARG, and NFKB1, when the heart was challenged by applied mechanical stress. Conclusions. This study unravels the complex transcriptome profiles of the heart tissues and highlighting the candidate genes involved in cardiac remodeling induced by mechanical stress may usher in a new era of precision diagnostics and treatment in patients with cardiac remodeling.

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