Transcriptomic signature of treatment with recombinant laminin 511-E8 and acetazolamide in human H69 cholangiocytes
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ABSTRACT:
PROVIDER: PRJNA915951 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR22892290_1.fastq.gz | Fastqsanger.gz | |||
| SRR22892290_2.fastq.gz | Fastqsanger.gz | |||
| SRR22892291_1.fastq.gz | Fastqsanger.gz | |||
| SRR22892291_2.fastq.gz | Fastqsanger.gz | |||
| SRR22892292_1.fastq.gz | Fastqsanger.gz |
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Project description:IgG4-related cholangitis (IRC) is the hepatobiliary manifestation of IgG4-related disease (IgG4-RD), a systemic, B-cell driven, fibroinflammatory disorder. To date, numerous autoantigens have been described including laminin 511-E8 and carbonic anhydrase (CA) enzymes (PMID 30089633, 15647194). Laminin 511-E8 is an extracellular matrix protein that has been shown to upregulate secretory components and aid in the cholangiocyte differentiation of induced pluripotent stem cells (PMID 27103433). Carbonic anhydrase enzymes catalyse the reversible hydration of carbon dioxide, thereby producing bicarbonate and a single proton. In humans there are 14 different CA isoforms with a confined subcellular distribution. All isoforms can be targeted by the pan-CA inhibitor acetazolamide, which is a registered drug for multiple indications. We aimed to investigate the role of IgG4-RD autoantigens laminin 511-E8 and CA enzymes in human cholangiocytes. Transcriptional profiling may provide insights into the underlying molecular mechanisms of these proteins on the cholangiocellular level. Thus, by RNA sequencing the transcriptomic signature of recombinant laminin 511-E8 and acetazolamide treatment was assessed in human H69 cholangiocytes.
2024-02-15 | GSE221746 | GEO
Project description:IgG4-related cholangitis (IRC) is the hepatobiliary manifestation of IgG4-related disease, a systemic B-cell-driven fibroinflammatory disorder. Four autoantigens have recently been described in IgG4-RD; annexin A11, galectin-3, laminin 511-E8, and prohibitin 1. We have previously reported a protective role of annexin A11 and laminin 511-E8 in human cholangiocytes against toxic bile acids. Here we explored a potentially protective role of the carbohydrate binding lectin galectin-3 and the scaffold proteins prohibitin 1 and 2.
2024-01-23 | PXD042856 | Pride
Project description:Some mouse embryonic stem cell (mESC) lines need to be maintained on feeder cells in gelatin/Std condition. To eliminate the need for feeder cells, we decided to maintain the C57BL6/J mESCs on dishes coated with Laminin-511 (LN511) enabling maintenance of mESCs without feeder cells in Std condition (Domogatskaya et al., 2008). To compare the transcriptomes of mESCs cultured on gelatin-coated dish and those cultured on Laminin-511 coated dish, we performed RNA-Seq analysis. We found that the transcriptomes of mESCs cultured on gelatin-coated dish and those cultured on LN511 coated dish showed no considerable difference in expression patterns.
2020-06-04 | GSE132591 | GEO
Project description:Simple Summary Numerous studies over the past few decades have revealed that the interactions of gastric cancer cells with laminins through integrins play important roles in tumor cell proliferation, infiltration, and metastasis. However, the association between gastric cancer cells and the laminin E8 fragment, which is the smallest integrin-binding component, has not been investigated. In this study, we revealed that the laminin 511-E8 fragment had a greater impact on the adhesion, morphology, and proliferation of gastric cancer cells than full-length laminin 511. Thus, the laminin 511-E8 fragment is considered to be suitable for investigating the interaction between gastric cancer cells and extracellular matrices in tumor invasion and metastasis. Further, the involvement of Cdc42 in the laminin 511-E8 fragment-induced enhanced adhesion of gastric cancer cells was suggested. Abstract Background: The interaction between cancer cells and laminin (Ln) is a key event in tumor invasion and metastasis. Previously, we determined the effect of full-length Ln511 on gastric cancer cells. However, the interactions between the Ln511-E8 fragment, a truncated protein of Ln511, and gastric cancer cells have not been investigated. Methods: We investigated the adhesion properties of gastric cancer cells to full-length Ln511 and Ln511-E8 fragments. Results: The proliferation of four gastric cancer cell lines (SH-10-TC, MKN74, SC-6-JCK, and MKN45) was highest on the Ln511-E8 fragment. Further, a larger cytoplasm was observed in SH-10-TC and MKN74 cells cultured on full-length Ln511 or Ln511-E8 fragments. The percentage of adhesive cells was highest on the Ln511-E8 fragment in all four cell lines. Moreover, adhesion of the gastric cancer cells to Ln511-E8 fragment-coated plates was reduced by the Cdc42 GTPase inhibitor in a dose-dependent manner, suggesting the involvement of Cdc42 in the Ln511-E8 fragment-induced enhanced adhesion of gastric cancer cells. Conclusions: The Ln511-E8 fragment had a greater impact on the adhesion, morphology, and proliferation of gastric cancer cells than full-length laminin. Thus, the Ln511-E8 fragment is suitable for investigating the interaction between gastric cancer cells and extracellular matrices in tumor invasion and metastasis.
| S-EPMC9164087 | biostudies-literature
Project description:H69 cells (human cholangiocytes) were exposed to TNF-alpha (10 ng/ml) for 8 h and were then collected for miRNA analysis, compared with non-treated control cells.
2011-01-01 | E-MEXP-2049 | biostudies-arrayexpress
Project description:Limbal melanocytes, located in the basal epithelial layer of the corneoscleral limbus, represent essential components of the corneal epithelial stem cell niche, but, due to difficulties in their isolation and cultivation, their biological roles and potential for stem cell-based tissue engineering approaches have not been comprehensively studied. Here, we established a protocol for the efficient isolation and cultivation of pure populations of human limbal melanocytes, which could be expanded at high yield by using recombinant laminin (LN)-511-E8 as culture substrate. Co-cultivation of limbal melanocytes with limbal epithelial stem/progenitor cells on fibrin hydrogels pre-incubated with LN-511-E8 resulted in multilayered stratified epithelial constructs within ten days. By reproducing physiological cell-cell and cell-matrix interactions of the native niche environment, these biomimetic co-culture systems provide a promising experimental model for investigating the functional roles of melanocytes in the limbal stem cell niche and their suitability for developing advanced epithelial grafts for ocular surface surface reconstruction.
| S-EPMC7338389 | biostudies-literature
Project description:Background & aimsIgG4-related cholangitis (IRC) is the hepatobiliary manifestation of IgG4-related disease. Anti-laminin 511-E8 autoantibodies have been identified in its pancreatic manifestation. Laminin 511-E8 promotes endothelial barrier function, lymphocyte recruitment, and cholangiocyte differentiation. Here, we investigate anti-laminin 511-E8 autoantibody presence in IRC, and mechanisms via which laminin 511 may contribute to cholangiocyte protection.MethodsAnti-laminin 511-E8 serum autoantibody positivity was assessed by ELISA. RNA sequencing and RT-qPCR were performed on human H69 cholangiocytes treated with recombinant laminin 511-E8. H69 cholangiocytes were subjected to shRNA knockdown targeting genes encoding laminin 511 (LAMA5, LAMB1, LAMC1) or treated with recombinant laminin 511-E8. Cholangiocellular bile acid influx was quantified radiochemically using 22,23-3H-glycochenodeoxycholic acid (GCDC). GCDC-induced apoptosis was determined by Caspase-3/7 assays. Cholangiocellular barrier function was assessed by FITC-Dextran permeability assays. Immunofluorescent staining of laminin 511 and claudin 1 was performed on extrahepatic bile duct tissue of control and anti-laminin 511-E8 positive individuals with IRC.ResultsSeven out of 52 individuals with IRC had autoantibodies against laminin 511-E8. Recombinant laminin 511-E8 led to differential expression of genes involved in secretion, barrier function, and inflammation. Knockdown of laminin 511 constituents increased toxic bile acid permeation and GCDC-induced apoptosis. Laminin 511-E8 treatment decreased toxic bile acid permeation and dose-dependently alleviated GCDC-induced apoptosis. LAMA5 and LAMC1 knockdown increased transepithelial permeability. Laminin 511-E8 treatment reduced transepithelial permeability and prevented T lymphocyte-induced barrier dysfunction. Laminin 511 and claudin 1 staining patterns appeared altered in anti-laminin 511-E8 positive individuals with IRC.ConclusionsLaminin 511-E8 is an autoantigen in subsets of individuals with IRC. Laminin 511 enhances cholangiocellular barrier function and protects cholangiocytes against T lymphocyte-induced barrier dysfunction, toxic bile acid permeation and bile acid-induced apoptosis.Impact and implicationsA subset of patients with IgG4-related cholangitis (IRC) has autoantibodies against laminin 511-E8. In human cholangiocytes, laminin 511 protects against (T lymphocyte-induced) epithelial barrier dysfunction and hydrophobic bile acids. Laminin 511 and claudin 1 staining may be altered in extrahepatic bile ducts of patients with IRC who are anti-laminin 511-E8 positive. This makes it tempting to speculate that a decreased epithelial barrier function with attraction of immune cells and impaired bicarbonate secretion as a result of dysfunction of laminin 511 by autoantibody binding could potentially be a common systemic pathogenic mechanism in a subset of patients with IgG4-RD.
| S-EPMC10959701 | biostudies-literature
Project description:The extracellular matrix plays a crucial role in stem cell maintenance, expansion and differentiation. Laminin, a basement membrane protein, is effective in supporting cultures of human pluripotent stem cells. To furhter investigate the effects of different isoforms of laminin on ectodemral differentiation of hiPSCs, we performed gene expression analysis of differentiated human iPS cells on E8 fragments of laminin isoforms (LN111E8, LN211E8, LN332E8, LN411E8, and LN511E8).
2018-10-17 | GSE116591 | GEO
Project description:To identify which gene expressions were affected by the different LM-E8 scaffolds, we performed transcriptome analysis using microarrays.
2021-04-21 | GSE149717 | GEO
Project description:Gene expression analysis of human pluripotent stem cells maintained on several laminin-E8 scaffolds.
| PRJNA629795 | ENA