scholarly journals PRDM1 controls the sequential activation of neural, neural crest and sensory progenitor determinants by regulating histone modification

2019 ◽  
Author(s):  
Ravindra S. Prajapati ◽  
Mark Hintze ◽  
Andrea Streit
Keyword(s):  
Neural Crest ◽  
Chromatin Immunoprecipitation ◽  
Specific Binding ◽  
Critical Role ◽  
Chromatin Modifications ◽  
Pluripotency Factors ◽  
Sequential Activation ◽  
Orderly Transition ◽  
Open Question

ABSTRACTDuring early embryogenesis, the ectoderm is rapidly subdivided into neural, neural crest and sensory progenitors. How the onset of lineage-specific determinants and the loss of pluripotency markers are temporally and spatially coordinated in vivo remains an open question. Here we identify a critical role for the transcription factor PRDM1 in the orderly transition from epiblast to defined neural lineages. Like pluripotency factors, PRDM1 is expressed in all epiblast cells prior to gastrulation, but lost as they begin to differentiate. We show that, unlike pluripotency factors, PRDM1 is initially required for the activation of neural, neural crest and sensory progenitor specifiers and for the downregulation of pluripotency-associated genes. In vivo chromatin immunoprecipitation reveals stage-specific binding of PRDM1 to regulatory regions of neural and sensory progenitor genes, PRDM1-dependent recruitment of the histone demethylase Kdm4a to these regions and associated removal of repressive histone marks. Once lineage determinants become expressed, they repress PRDM1, and our data suggest that PRDM1 downregulation is required for cells to maintain their identity. Thus, PRDM1 mediates chromatin modifications that directly control neural and sensory progenitor genes, and its activities switch from an activator at early stages to a repressor once neural fates have been established.

scholarly journals A PHD12–Snail2 repressive complex epigenetically mediates neural crest epithelial-to-mesenchymal transition

2012 ◽  
Vol 198 (6) ◽  
pp. 999-1010 ◽  
Author(s):  
Pablo H. Strobl-Mazzulla ◽  
Marianne E. Bronner
Keyword(s):  
Neural Crest ◽  
Adhesion Molecule ◽  
Chromatin Immunoprecipitation ◽  
Epithelial To Mesenchymal Transition ◽  
Adaptor Protein ◽  
Unknown Mechanism ◽  
Mesenchymal Transition ◽  
Histone 3 ◽  
Active Transcription

Neural crest cells form within the neural tube and then undergo an epithelial to mesenchymal transition (EMT) to initiate migration to distant locations. The transcriptional repressor Snail2 has been implicated in neural crest EMT via an as of yet unknown mechanism. We report that the adaptor protein PHD12 is highly expressed before neural crest EMT. At cranial levels, loss of PHD12 phenocopies Snail2 knockdown, preventing transcriptional shutdown of the adhesion molecule Cad6b (Cadherin6b), thereby inhibiting neural crest emigration. Although not directly binding to each other, PHD12 and Snail2 both directly interact with Sin3A in vivo, which in turn complexes with histone deacetylase (HDAC). Chromatin immunoprecipitation revealed that PHD12 is recruited to the Cad6b promoter during neural crest EMT. Consistent with this, lysines on histone 3 at the Cad6b promoter are hyperacetylated before neural crest emigration, correlating with active transcription, but deacetylated during EMT, reflecting the repressive state. Knockdown of either PHD12 or Snail2 prevents Cad6b promoter deacetylation. Collectively, the results show that PHD12 interacts directly with Sin3A/HDAC, which in turn interacts with Snail2, forming a complex at the Cad6b promoter and thus revealing the nature of the in vivo Snail repressive complex that regulates neural crest EMT.

scholarly journals CircDUSP16 promotes the tumorigenesis and invasion of gastric cancer by sponging miR-145-5p

2019 ◽  
Vol 23 (3) ◽  
pp. 437-448 ◽  
Author(s):  
Zizhen Zhang ◽  
Chaojie Wang ◽  
Yeqian Zhang ◽  
Site Yu ◽  
Gang Zhao ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Specific Binding ◽  
Critical Role ◽  
Ectopic Expression ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Poor Survival ◽  
Data Set

Abstract Background Circular RNAs (circRNAs) as a novel subgroup of non-coding RNAs act a critical role in the pathogenesis of gastric cancer (GC). However, the underlying mechanisms by which hsa_circ_0003855 (circDUSP16) contributes to GC are still undocumented. Materials The differentially expressed circRNAs were identified by GEO database. The association of circDUSP16 or miR-145-5p expression with clinicopathological features and prognosis in GC patients was analyzed by FISH and TCGA-seq data set. Loss- and gain-of-function experiments as well as a xenograft tumor model were performed to assess the role of circDUSP16 in GC cells. circDUSP16-specific binding with miR-145-5p was confirmed by bioinformatic analysis, luciferase reporter, and RNA immunoprecipitation assays. Results The expression levels of circDUSP16 were markedly increased in GC tissue samples and acted as an independent prognostic factor of poor survival in patients with GC. Knockdown of circDUSP16 repressed the cell viability, colony formation, and invasive potential in vitro and in vivo, but ectopic expression of circDUSP16 reversed these effects. Moreover, circDUSP16 possessed a co-localization with miR-145-5p in the cytoplasm, and acted as a sponge of miR-145-5p, which attenuated circDUSP16-induced tumor-promoting effects and IVNS1ABP expression in GC cells. MiR-145-5p had a negative correlation with circDUSP16 expression and its low expression was associated with poor survival in GC patients. Conclusions CircDUSP16 facilitates the tumorigenesis and invasion of GC cells by sponging miR-145-5p, and may provide a novel therapeutic target for GC.

scholarly journals In Vivo Regulation of Follicle-Stimulating Hormone Receptor by the Transcription Factors Upstream Stimulatory Factor 1 and Upstream Stimulatory Factor 2 Is Cell Specific

Endocrinology ◽  
2008 ◽  
Vol 149 (10) ◽  
pp. 5297-5306 ◽  
Author(s):  
Brian P. Hermann ◽  
Kaori Hornbaker ◽  
Daren A. Rice ◽  
Michele Sawadogo ◽  
Leslie L. Heckert
Keyword(s):  
Granulosa Cells ◽  
Sertoli Cells ◽  
Chromatin Immunoprecipitation ◽  
Leucine Zipper ◽  
Specific Binding ◽  
Pubertal Timing ◽  
Upstream Stimulatory Factor ◽  
Stimulatory Factor

Pituitary FSH promotes pubertal timing and normal gametogenesis by binding its receptor (FSHR) located on Sertoli and granulosa cells of the testis and ovary, respectively. Studies on Fshr transcription provide substantial evidence that upstream stimulatory factor (USF) 1 and USF2, basic helix-loop-helix leucine zipper proteins, regulate Fshr through an E-box within its promoter. However, despite the strong in vitro support for USF1 and USF2 in Fshr regulation, there is currently no in vivo corroborating evidence. In the present study, chromatin immunoprecipitation demonstrated specific binding of USF1 and USF2 to the Fshr promoter in both Sertoli and granulosa cells, in vivo. Control cells lacking Fshr expression showed no USF-Fshr promoter binding, thus correlating USF-promoter binding to gene activity. Evaluation of Fshr expression in Usf1 and Usf2 null mice further explored USF’s role in Fshr transcription. Loss of either gene significantly reduced ovarian Fshr levels, whereas testis levels were unaltered. Chromatin immunoprecipitation analysis of USF-Fshr promoter binding in Usf-null mice indicated differences in the composition of promoter-bound USF dimers in granulosa and Sertoli cells. Promoter-bound USF dimer levels declined in granulosa cells from both null mice, despite increased USF2 levels in Usf1-null ovaries. However, compensatory increases in promoter-bound USF homodimers were evident in Usf-null Sertoli cells. In summary, this study provides the first in vivo evidence that USF1 and USF2 bind the Fshr promoter and revealed differences between Sertoli and granulosa cells in compensatory responses to USF loss and the USF dimeric composition required for Fshr transcription.

Conditional Expression of C/EBPα in Mice Reveals Developmental Plasticity of Lymphoid and Erythroid/ Megakaryocyte Precursors in Vivo.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 671-671 ◽  
Author(s):  
Yumi Fukuchi ◽  
Fumi Shibata ◽  
Miyuki Ito ◽  
Yuko Goto-Koshino ◽  
Yusuke Sotomaru ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Developmental Plasticity ◽  
Specific Binding ◽  
Critical Role ◽  
Ectopic Expression ◽  
Pcr Analysis ◽  
Lineage Specification ◽  
Myeloid Lineage ◽  
Conditional Expression

Abstract CCAAT/ enhancer binding protein α (C/EBP α) is a member of the bZIP family of transcription factors that plays a critical role for early myeloid differentiation. C/EBP α knockout mice showed a complete differentiation block at myeloblast stage in hematopoietic system, and mature neutrophils and eosinophils are absent in the peripheral blood. Lineage specification in developmental tree of hematopoiesis is generally determined by a lineage specific transcription factors such as C/EBP α and GATA-1 that allow commitment to the specific lineage with simultaneous extinction of their capacity to differentiate into the other ones. However, recent evidences shown by the ectopic expression of above transcription factors unveiled the unexpected developmental plasticity of various progenitors such as MEP (erythroid/ megakaryocyte progenitor) and CLP (common lymphoid progenitor). GATA-1 is reported to convert CLP and CMP (common myeloid progenitor) into erythroid/ megakaryocyte lineage, however, the effect of C/EBP α on MEP and CLP is still unlcear. In order to investigate the role of C/EBP α in the various aspect of hematopoietic differentiation, especially its effect on the lineage specification at different stages of differentiation in vivo, we generated transgenic mice expressing inducible form of C/EBP α (C/EBP α-ER) under H-2K promoter (C/EBP α-ER Tg). In these mice, C/EBP α activity can be induced conditionally by 4-hydroxy tamoxifen (4-HT) in all hematopoietic cells. As expected, C/EBP α-ER was expressed in almost all hematopoietic tissues including bone marrow, spleen and thymus in these mice. Gel shift analysis revealed that C/EBP α-ER was activated by 4-HT, and showed specific binding to C/EBP-specific oligonucleotide in these tissues. Next we tested differentiation plasticity of erythroid and lymphoid progenitors by ectopically inducing C/EBP α-ER activity in these cells. We sorted MEP and CLP by FACS from C/EBP α-ER Tg and examined their clonogenic activities in the presence or absence of 4-HT. In the absence of 4-HT, MEP and CLP exclusively formed erythroid/ megakaryocyte and lymphoid colonies, respectively, as previously reported. Surprisingly however, these cells dramatically changed their fate of differentiation and formed significant numbers of granulocyte/ macrophage (GM) colonies in the presence of 4-HT, indicating that ectopic activation of C/EBP α-ER activity skewed their differentiation pathways to myeloid lineage. Cytospin preparation of the colonies and RT-PCR analysis revealed that these were accompanied by the morphological differentiation to granulocytes/ macrophages, and upregulation of myeloid-specific genes at mRNA level. These results indicate that MEP and CLP are not fully committed to either erythroid/ megakaryocyte or lymphoid lineage, and possess differentiation plasticity that can be redirected to myeloid lineage.

scholarly journals Long Noncoding RNA uc.412 Promotes Mesangial Cell Fibrosis via Binding ELAVL1 in Glomerulosclerosis

2020 ◽  
Author(s):  
Jialing Ji ◽  
Ju Rong ◽  
Ming Sun ◽  
Chan Huang ◽  
Jin Wang ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Noncoding Rna ◽  
Mesangial Cells ◽  
Specific Binding ◽  
Critical Role ◽  
Convincing Evidence ◽  
Tgf Β1

Abstract Background: Glomerulosclerosis is a characteristic pathologic feature in chronic kidney disease (CKD). Convincing evidence indicates that the mesangial cells (MCs) play critical role in this process. However, the exact mechanism remains unclear. Using RNA-seq analysis, we previously found that lncRNA uc.412 was involved in the MC proliferation. Here, the effect of uc.412 on glomerular fibrosis and the potential mechanism were explored. Methods: In vivo, CKD mice models were established by 5/6 nephrectomy. The expression of lncRNA uc.412 in CKD was detected by Real-Time PCR. In vitro, MCs were intervened with TGF-β1 (10ng/mL). The uc.412 expression in MCs was detected by in site hybridization. MCs were transfected with uc.412 siRNA or a lentivirus targeting uc.412 and then examined using western blot, Real-Time PCR, RNA pull down assay and immunofluorescence staining. Results: We found that the expression of uc.412 was significantly increased in CKD mice and is induced by TGF-β1 via Smad3- dependent signal pathway. Overexpressing uc.412 caused MCs fibrosis and knockdown of uc.412 alleviated TGF-β1-induced MCs fibrosis. Using RNA pull down analysis, we found that the ELAVL1 was the specific binding protein for uc.412. Moreover, ELAVL1 expression was increased in TGF-β1-treated MCs and silencing ELAVL1 expression attenuated MCs fibrosis. Conclusions: Thus, here, we demonstrated that uc.412, which is regulated in a Smad3-dependent mechanism, is significantly increased during progression of CKD via regulating ELAVL1 expression. Our findings provided the therapeutic strategy for treatment of CKD.

scholarly journals Autocrine Production of Interleukin-34 Promotes the Development of Endometriosis through CSF1R/JAK3/STAT6 signaling

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Kaiqing Lin ◽  
Junyan Ma ◽  
Yaomin Peng ◽  
Meina Sun ◽  
Kaihong Xu ◽  
...  
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Critical Role ◽  
Neutralizing Antibody ◽  
Migration And Invasion ◽  
Endometrial Stromal Cells ◽  
Novel Therapeutic Strategies ◽  
Stimulating Factor ◽  
Insight Into

AbstractInterleukin (IL)-34 plays a critical role in cell proliferation, differentiation, apoptosis, angiogenesis, inflammation and immunoregulation. Numerous diseases can be attributed to the dysregulation of IL-34 signaling. This study was performed to investigate the function of IL-34 in the pathogenesis of endometriosis. Firstly, by enzyme linked immunoabsorbent assay, we found that IL-34, VEGF, MMP-2 and MMP-9 were increased in the sera of patients with endometriosis. Secondly, exposure to IL-34 promoted the proliferation, migration and invasion of eutopic endometrial stromal cells (ESCs). Additionally, stimulation with IL-34 up-regulated colony-stimulating factor 1 receptor (CSF1R), p-JAK3, p-STAT6, VEGF, MMP-2 and MMP-9 in these eutopic ESCs. Treatment with AS1517499, an inhibitor of STAT6, remarkably abrogated the alterations induced by IL-34. A Chromatin immunoprecipitation (ChIP) assay demonstrated binding of STAT6 to the IL-34 promoter, further implicating STAT6 in IL-34 signaling. Notably, reverse results were obtained in ectopic ESCs with the application of an IL-34 neutralizing antibody. In vivo, AS1517499 suppressed the maintenance of endometriosis lesions in rats. In summary, autocrine production of IL-34, mediated by STAT6, promoted the development of endometriosis in vitro and in vivo through the CSF1R/JAK3/STAT6 pathway. Our research reveals the function of IL-34 in endometriosis, which may provide insight into novel therapeutic strategies for endometriosis.

scholarly journals Histone demethylase KDM4B regulates otic vesicle invagination via epigenetic control of Dlx3 expression

2015 ◽  
Vol 211 (4) ◽  
pp. 815-827 ◽  
Author(s):  
Rosa A. Uribe ◽  
Ailín L. Buzzi ◽  
Marianne E. Bronner ◽  
Pablo H. Strobl-Mazzulla
Keyword(s):  
Inner Ear ◽  
Histone Methylation ◽  
Chromatin Immunoprecipitation ◽  
Morphological Changes ◽  
Critical Role ◽  
Histone Demethylase ◽  
Otic Vesicle ◽  
Direct Target ◽  
Otic Placode

In vertebrates, the inner ear arises from the otic placode, a thickened swathe of ectoderm that invaginates to form the otic vesicle. We report that histone demethylase KDM4B is dynamically expressed during early stages of chick inner ear formation. A loss of KDM4B results in defective invagination and striking morphological changes in the otic epithelium, characterized by abnormal localization of adhesion and cytoskeletal molecules and reduced expression of several inner ear markers, including Dlx3. In vivo chromatin immunoprecipitation reveals direct and dynamic occupancy of KDM4B and its target, H3K9me3, at regulatory regions of the Dlx3 locus. Accordingly, coelectroporations of DLX3 or KDM4B encoding constructs, but not a catalytically dead mutant of KDM4B, rescue the ear invagination phenotype caused by KDM4B knockdown. Moreover, a loss of DLX3 phenocopies a loss of KDM4B. Collectively, our findings suggest that KDM4B play a critical role during inner ear invagination via modulating histone methylation of the direct target Dlx3.

Implications of glycoprotein VI for theranostics

2014 ◽  
Vol 112 (07) ◽  
pp. 26-31 ◽  
Author(s):  
Sebastian Vogel ◽  
Christina Pfannenberg ◽  
Bernd Pichler ◽  
Harald Langer ◽  
Boris Bigalke ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Cardiovascular Diseases ◽  
Specific Binding ◽  
Thrombus Formation ◽  
Critical Role ◽  
Vascular Lesions ◽  
Soluble Form ◽  
Glycoprotein Vi ◽  
Non Invasive

SummaryGlycoprotein VI (GPVI), a membrane glycoprotein solely expressed in platelets and megakaryocytes, plays a critical role in thrombus formation due to collagen/GPVI-mediated platelet activation and adhesion. Recent studies have shown that surface expression of GPVI on circulating platelets is enhanced in acute cardiovascular diseases such as myocardial infarction and ischaemic stroke. Increased GPVI levels are associated with poor clinical outcome and are an early indicator for imminent myocardial infarction in patients with chest pain. The soluble form of the dimeric GPVI fusion protein (sGPVI-Fc) binds with high affinity to collagen and atherosclerotic plaque tissue. Non-invasive imaging studies with radiolabelled sGPVI-Fc show specific binding activity to vascular lesions in vivo. Further, sGPVI-Fc has been developed as a new therapeutic platelet-based strategy for lesion-directed antithrombotic therapy. This review summarises the potential of GPVI for diagnostic and therapeutic options based on novel non-invasive molecular imaging modalities to ameliorate care of patients with cardiovascular diseases.

EPCO-05. GENOME-WIDE ANALYSIS OF TEAD1 OCCUPANCY IN BIOLOGICALLY DISTINCT GLIOBLASTOMA SAMPLES

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi2-vi2
Author(s):  
Tanvi Joshi ◽  
German Nudelman ◽  
Elena Zaslavsky ◽  
Nadejda Tsankova
Keyword(s):  
Cell Biology ◽  
Target Genes ◽  
Specific Binding ◽  
Treatment Options ◽  
Critical Role ◽  
Negative Control ◽  
Functional Enrichment ◽  
Rapid Progression

Abstract The diffusely infiltrative nature of glioblastoma (GBM) cells is a major contributor to the disease’s aggressive behavior, including its rapid progression and therapeutic resistance. Moreover, current treatment options do not target the invasive nature of GBM. Recent chromatin accessibility studies prioritized enrichment of the TEAD1 transcription factor motif in glioblastoma stem cell biology and subsequent knockout and overexpression studies confirmed a critical role for TEAD1 in GBM migration, in vitro and in vivo. However, the downstream mechanisms through which TEAD1 regulates GBM cell migration remain poorly understood. In this study, we performed chromatin immunoprecipitation (ChIP-seq) using TEAD1-specific antibody and IgG as non-specific binding control, to characterize TEAD1 occupancy across GBM samples with unique genomic alterations. ChIP-seq peaks were called using MACS2, filtered for duplicates and blacklisted regions, and normalized per sample to their respective genomic input. Initial functional enrichment analyses were performed on three GBM samples with the highest number of TEAD1 occupancy peaks using CistromeGO, which ranked genes based on their TEAD1-specific regulatory potential (RP) score, as a function of peak number and distance from the transcription start site. Analyses of the top 1000 genes with highest TEAD1 RP scores identified 132 common targets across all samples, including known TEAD target genes ETV1 and Cyr61, which related to angiogenesis, cadherin and integrin signaling, cell adhesion, and chromatin regulation gene ontology terms, among others. Interestingly, KEGG pathway analysis also revealed Hippo pathway enrichment across all GBM samples, suggesting a possible TEAD1 regulatory feedback loop in GBM. Analysis of TEAD1 ChIP-seq peaks in non-GBM negative control tissue did not show functional enrichment for any of the terms seen in the GBM samples. Ongoing analyses are focused on characterizing TEAD1 occupancy at active cis-regulatory regions using parallel H3K27ac ChIP-seq data, in order to prioritize the most salient TEAD1-regulatory targets in GBM.

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