CREPT aggravates liver fibrosis through TGF-β activated-Smad2/3 but not Smad4 in hepatocytes
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1310511 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:Background & Aims: Liver fibrosis, a serious medical condition with limited therapies, frequently progresses to cirrhosis and other severe complications. Here, we report that cell cycle-related and expression elevated protein in tumor (CREPT) is up-regulated in liver fibrosis. CREPT preferentially enhances transforming growth factor-β (TGF-β)-activated Smad2/3, rather than the canonical Smad2/3-Smad4 complex, thereby facilitating liver fibrosis progression. Methods: We analyzed public transcriptomic and proteomic datasets from patients with liver fibrosis. Human tissue arrays and mouse models of fibrosis were used to investigate CREPT expression patterns. Hepatocyte-specific overexpression (TG-CREPT) and deletion (CREPThep-/-) mice were subjected to acute liver injury (24 h, high-dose CCl4), chronic fibrosis (28 d, low-dose CCl4), and bile duct ligation (BDL) to induce liver fibrosis. Histological, biochemical, and RNA-sequencing (RNA-seq) analyses and CUT&Tag experiments were conducted. Results: CREPT is predominantly enriched in hepatocytes and upregulated in fibrotic liver tissues from patients and mice induced by CCl4 or BDL. Overexpression of CREPT in hepatocytes exacerbates liver injury and fibrosis in mice under acute or chronic CCl4 exposure and BDL challenge. In contrast, hepatocyte-specific deletion of CREPT significantly alleviates liver fibrosis. Mechanistically, CREPT aggravates TGF-β/Smad signaling pathway, preferentially interacting with Smad2/3, but not Smad4, to promote inflammatory responses, and up-regulates fibrogenic genes, including Cebpd, Fosl2 and Fn1. Intriguingly, targeting CREPT with LNP-delivered siRNA (LNP-siCREPT) and GalNAc-delivered siRNA (GalNAc-siCREPT) significantly reduces pre-existing liver fibrosis in mice. Conclusions: Our findings identify a promoting effect of CREPT on liver fibrosis through TGF-β-activated Smad2/3 but not Smad4. Targeting CREPT represents a potential therapeutic strategy for liver fibrosis.
2025-08-28 | GSE306355 | GEO
Project description:TGF-β signaling is known to be very much dependent on the formation of Smad2/3-Smad4 transcription regulatory complexes. However, the signaling functions of Smad2/3-Smad4 in TGF-β-induced responses are obscure as TGF-β also initiates a number of other signaling pathways. In this study, we systematically assessed the contribution of TGF-β-Smad2/3-Smad4 signaling to target gene transcription. Individual Smads were selectively knocked down in Hep3B cells by stable RNA interference (RNAi). We identified TGF-β-responsive genes using genome-wide oligonucleotide microarrays and confirmed their dependency on Smad2, Smad3 or Smad4 by the combination of RNAi and microarray assay. The major finding from our microarray analysis was that of the 2039 target genes seen to be regulated via TGF-β induction, 190 were differentially transcriptionally controlled by Smad2-Smad4 and Smad3-Smad4 signaling and the latter control mechanism appeared to be functionally more important. We also found evidence of competition between Smad2 and Smad3 for their activation when controlling the transcription of target genes. Keywords: cell type comparison
2006-11-14 | GSE6242 | GEO
Project description:The tumor suppressive effects of TGF-β are classically associated with the activation of the “canonical” SMAD-mediated pathway, whereas its oncogenic effects are largely attributed to its “non-canonical signaling”. We herein provide evidence of an oncogenic effect for SMAD2 and 3 in response to TGF-β in SMAD4-null cancer cells. Using the CRISPR/Cas9 technology, we report that simultaneous knockout of Smad2 and 3 in Smad4-negative pancreatic ductal adenocarcinoma (PDAC) cells compromises TGF-β-driven collective migration mediated by FAK and Rho/Rac signaling. Moreover, RNA-sequencing analyses highlight a TGF-β gene signature related to aggressiveness mediated by SMAD2 and 3 in the absence of SMAD4. Using PDAC patients cohorts, we reveal that SMAD4-negative tumors with high levels of (phospho)-SMAD2 are more aggressive and have a poorer prognosis. Thus, loss of SMAD4 tumor suppressive activity in PDAC is associated with oncogenic gain-of-function of SMAD2 and 3 and the onset of associated deleterious effects.
2022-10-09 | GSE178714 | GEO
Project description:The FOXL2 mutant C134W occurs in virtually all adult ovarian granulosa cell tumors (AGCT), and is considered a driver of oncogenesis in this disease. However, the mechanism by which FOXL2C134W contributes to tumorigenesis is not known. Here, we show that mutant FOXL2C134W acquires the ability to bind SMAD4, forming a FOXL2C134W/SMAD4/SMAD2/3 complex that binds a novel hybrid DNA motif AGHCAHAA, unique to the FOXL2C134W mutant. This binding induces an enhancer-like chromatin state leading to transcription of nearby genes, many of which are characteristic of stemness and epithelial-to-mesenchymal transition. Importantly, primary AGCT tumors display a strong FOXL2C134W enrichment at hybrid loci. Ablation of SMAD4 or SMAD2/3 inhibits FOXL2C134W binding at hybrid sites and decreases transcription of associated genes. Accordingly, TGFβ-inhibition mitigates the transcriptional effect of FOXL2C134W.
2020-07-01 | GSE138496 | GEO
Project description:To dissected the roles of SMAD2/3 and SMAD4 in mouse embryonic stem cells (mESC), knockouts of SMAD2/3 (SMAD2 and SMAD3 double knockout, S2/3DKO) and SMAD4 (SMAD4 knockout, S4KO) were used to conduct RNAseq in ES and EB conditions.
2024-10-21 | GSE240785 | GEO
Project description:To understand the genome wide binding profile of SMAD2/3 when SMAD4 is lost, Smad2/3 ChIPseq was performed in SMAD4 knockout (S4KO) mouse embryonic stem cells (mESC) or in S4KO embryonic bodies (EBs). To figure out the effect of Dnmt3b on SMAD2/3 binding ability in the absence of SMAD4, we further performed SMAD2/3 CUT&Tag in EBs of S4KO and S4KO with Dnmt3b konckdown, as well as in mesendoderm (ME) stage of WT and S4KO. The binding sites of Dnmt3a and Dnmt3b were explored by Dnmt3a or Dnmt3b ChIP-seq and Dnmt3b CUT&Tag in WT EBs and S4KO EBs. WT with Dnmt3b knockdown and S4KO with Dnmt3b knockdown were used to as a control for the Dnmt3b CUT&Tag. Besides, we inserted a 3xFlag-tag at the C-terminus of Dnmt3a or Dnmt3b and then carried out Flag CUT&Tag in EBs.
2024-10-21 | GSE240786 | GEO
Project description:SMAD2/3 mediate oncogenic effects of TGF-β in the absence of SMAD4
| PRJNA740178 | ENA
Project description:The transforming growth factor beta (TGFβ) related signaling is one of the most important signaling pathways regulating early developmental events. Smad2 and Smad3 are structurally similar and it is mostly considered that they are equally important in mediating TGFβ signals. Here, we show that Smad3 is an insensitive TGFβ transducer as compared with Smad2. Smad3 preferentially localizes within the nucleus and is thus sequestered from membrane signaling. The ability of Smad3 in oligomerization with Smad4 upon agonist stimulation is also impaired given its unique linker region. Smad2 mediated TGFβ signaling plays a crucial role in epiblast development and patterning of three germ layers. However, signaling unrelated nuclear localized Smad3 is dispensable for TGFβ signaling-mediated epiblast specification, but important for early neural development, an event blocked by TGFβ/Smad2 signaling. Both Smad2 and Smad3 bind to the conserved Smads binding element (SBE), but they show nonoverlapped target gene binding specificity. We conclude that Smad2 and Smad3 possess differential sensitivities in relaying TGFβ signaling and have distinct roles in regulating early developmental events. GFP, GFP-Smad2 and GFP-Smad3 constitutively expressed Smad3-/- mouse ESCs were differentiated to day6 neuroepithelia and collected for Chip-Seq with an anti-GFP antibody.
2016-03-01 | E-GEOD-76557 | biostudies-arrayexpress
Project description:The vertebrate homologues of Drosophila dachsund, DACH1 and DACH2, have been implicated as important regulatory genes in development. DACH1 plays a role in retinal and pituitary precursor cell proliferation and DACH2 plays a specific role in myogenesis. DACH proteins contain a domain (DS-domain) that is conserved with the proto-oncogenes Ski and Sno. Since the Ski/Sno proto-oncogenes repress AP-1 and SMAD signaling, we hypothesized that DACH1 might play a similar cellular function. Herein, DACH1 was found to be expressed in breast cancer cell lines and to inhibit TGF-beta induced apoptosis. DACH1 repressed TGF-beta induction of AP-1 and Smad signaling in gene reporter assays and repressed endogenous TGF-beta responsive genes by microarray analyses. DACH1 bound to endogenous NCoR and Smad4 in cultured cells and DACH1 co-localized with NCoR in nuclear dot-like structures. NCoR enhanced DACH1 repression and the repression of TGF-beta-induced AP-1 or Smad-signaling by DACH1 required the DACH1 DS domain. The DS-domain of DACH was sufficient for NCoR-binding at a Smad4-binding site. Smad4 was required for DACH1 repression of Smad signaling. In Smad4 null HTB-134 cells, DACH1 inhibited the activation of SBE-4 reporter activity induced by Smad2 or Smad3 only in the presence of Smad4. DACH1 participates in the negative regulation of TGF-beta signaling by interacting with NCoR and Smad4. Keywords: other
2003-10-15 | GSE685 | GEO
Project description:CREPT was demonstrated as a key regulator of intestinal stem cells. To figure out the role of CREPT on transcription level, a ChIP-seq experiment was performed to identify the CREPT down-stream genes in intestinal crypt cells.
2020-11-08 | GSE158243 | GEO