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Transcriptomic profiling of three-dimensional cholangiocyte spheroids long-term exposed to repetitive Clonorchis sinensis excretory-secretory products

ABSTRACT: Microarray and RNA-Seq analysis revealed that ESP-treated cholangiocyte H69 spheroids displayed global changes in gene expression compared to untreated spheroids. In ESP-treated H69 spheroids, 185 and 63 probes were found to be significantly upregulated and downregulated, respectively, corresponding to 209 genes (p < 0.01, fold change > 2).

ORGANISM(S): Homo sapiens

PROVIDER: GSE168792 | GEO | 2021/04/27

REPOSITORIES: GEO

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Transcriptomic profiling of three-dimensional cholangiocyte spheroids long term exposed to repetitive Clonorchis sinensis excretory-secretory products.

Project description:BackgroundBiliary tract infection with the carcinogenic human liver fluke, Clonorchis sinensis, provokes chronic inflammation, epithelial hyperplasia, periductal fibrosis, and even cholangiocarcinoma. Complications are proportional to the intensity and duration of the infection. In addition to mechanical irritation of the biliary epithelia from worms, their excretory-secretory products (ESPs) cause chemical irritation, which leads to inflammation, proliferation, and free radical generation.MethodsA three-dimensional in vitro cholangiocyte spheroid culture model was established, followed by ESP treatment. This allowed us to examine the intrinsic pathological mechanisms of clonorchiasis via the imitation of prolonged and repetitive in vivo infection.ResultsMicroarray and RNA-Seq analysis revealed that ESP-treated cholangiocyte H69 spheroids displayed global changes in gene expression compared to untreated spheroids. In ESP-treated H69 spheroids, 185 and 63 probes were found to be significantly upregulated and downregulated, respectively, corresponding to 209 genes (p < 0.01, fold change > 2). RNA-Seq was performed for the validation of the microarray results, and the gene expression patterns in both transcriptome platforms were well matched for 209 significant genes. Gene ontology analysis demonstrated that differentially expressed genes were mainly classified into immune system processes, the extracellular region, and the extracellular matrix. Among the upregulated genes, four genes (XAF1, TRIM22, CXCL10, and BST2) were selected for confirmation using quantitative RT-PCR, resulting in 100% similar expression patterns in microarray and RNA-Seq.ConclusionsThese findings broaden our understanding of the pathological pathways of liver fluke-associated hepatobiliary disorders and suggest a novel therapeutic strategy for this infectious cancer.

| S-EPMC8056535 | biostudies-literature

Human cholangiocarcinoma cells treated with Clonorchis sinensis excretory-secretory products.

Project description:Clonorchiasis is associated with bile duct malignancy and the subsequent development of cholangiocarcinoma. Although this is likely caused by adult Clonorchis sinensis and its excretory-secretory products (ESP), the precise molecular mechanisms remain obscure. To evaluate the effect of C. sinensis infection on differential gene expression in host hepatocytes, we used cDNA microarrays in human cholangiocarcinoma cells through the mimicking of C. sinensis infestation, and analyzed differential mRNA expression patterns of host cells. Keywords: Time course

2008-07-24 | GSE12213 | GEO

Transcriptomic profiling of three-dimensional cholangiocyte spheroids long-term exposed to repetitive Clonorchis sinensis excretory-secretory products

Project description:Transcriptomic profiling of three-dimensional cholangiocyte spheroids long-term exposed to repetitive Clonorchis sinensis excretory-secretory products

| PRJNA714054 | ENA

Gene expression profile at single cell level of human peripheral blood mononuclear cells cocultured with rotavirus treated or PBS treated human cholangiocyte cell line H69.

Project description:Biliary atresia (BA) is an devastating pediatric cholangiopathy and the leading indication for liver transplant in children worldwide. An in vitro cell system for investigating BA is still lacking. We develop an in vitro cell culture system to mimic immune dysfunction of biliary atresia by coculturing human peripheral blood mononuclear cells (PBMC) with rotavirus treated or PBS treated human cholangiocyte cell line H69. Single cell RNA sequencing analysis of PBMC after coculture was applied to validate the efficacy and fidelity of cell system.

2024-05-15 | GSE225177 | GEO

Cholangiocyte organoid culture in hydrogels from decellularized liver tissue

Project description:Culture of human cholangiocyte organoids in hydrogels derived from healthy liver extracellular matrix (LECM) extracts prepared from decellularized human livers are evaluated in an effort to establish a platform for production of cholangiocyte organoids for clinical regenerative applications. Human intrahepatic cholangiocyte organoids (ICO) grown in hydrogels made from LECM are compared those grown in mouse tumor derived basement membrane extracts (BME). Culture was performed with amino acids labeled with stable heavy isotopes to enable separation of ECM from the hydrogels from that of the produced by the cells with mass spectrometry (MS). MS data were used to evaluate the protein production of ICO comparing the different hydrogel substrates. The study also contains evaluation of the properties of the hydrogel substrates and focuses on expansion and differentiation of the ICO.

2022-04-13 | PXD028417 | Pride

Clonorchis sinensis transcriptome

Project description:We conducted transcriptome sequencing of Clonorchis sinesis sub-organs (sucker, muscle, ovary and testis) with Illumina sequencing.

2013-01-30 | E-MTAB-827 | biostudies-arrayexpress

Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components.

Project description:Clonorchis sinensis is a carcinogenic human liver fluke that promotes hepatic inflammatory environments via direct contact or through their excretory-secretory products (ESPs), subsequently leading to cholangitis, periductal fibrosis, liver cirrhosis, and even cholangiocarcinoma (CCA). This study was conducted to examine the host inflammatory responses to C. sinensis ESPs and their putative protein components selected from C. sinensis expressed sequenced tag (EST) pool databases, including TGF-β receptor interacting protein 1(CsTRIP1), legumain (CsLeg), and growth factor binding protein 2 (CsGrb2). Treatment of CCA cells (HuCCT1) with the ESPs or bacterial recombinant C. sinensis proteins differentially promoted the secretion of proinflammatory cytokines (IL-1β, IL-6, and TNF-α) as well as anti-inflammatory cytokines (IL-10, TGF-β1, and TGF-β2) in a time-dependent manner. In particular, recombinant C. sinensis protein treatment resulted in increase (at maximum) of ~7-fold in TGF-β1, ~30-fold in TGF-β2, and ~3-fold in TNF-α compared with the increase produced by ESPs, indicating that CsTrip1, CsLeg, and CsGrb2 function as strong inducers for secretion of these cytokines in host cells. These results suggest that C. sinensis ESPs contribute to the immunopathological response in host cells, leading to clonorchiasis-associated hepatobiliary abnormalities of greater severity.

| S-EPMC6753296 | biostudies-literature

MicroRNA profiling of human cholangiocytes in response to interferon-gamma

Project description:Cells of a human cholangiocyte cell line (H69) were exposed to 10 ng/ml of IFN-gamma for 8 h. Cells were collected and processed for miRNA array analysis carried out by EXIQON (Denmark).

2009-02-10 | E-MEXP-1906 | biostudies-arrayexpress

Transcription profiling of undifferentiated human extrahepatic cholangiocyte organoids (ECOs)

Project description:3D cell culture systems (organoids), cultured from extrahepatic bile duct biopsies, resemble biliary epthelium (cholangiocytes) upon culture according to the protocol previously established for culturing intrahepatic cholangiocyte organoids from liver biopsies (Huch et al. Cell 2015). These extrahepatic cholangiocyte organoids (ECOs) in Canonical-Wnt culture conditions maintain their genetic stability, and transcription profile and can be maintained and propagated during long periods of culturing while still maintaining their functional properties. The data presented in this ArrayExpress submission contained micro array results of 3 different ECO lines that were analyzed for their functional cholangiocyte ion-channels, involved in secreting a protective bicarbonate layer. It was shown that these data closely resemble the expression profile of intrahepatic cholangiocyte organoids (ICOs) as described by our group. Results of the ICOs are deposited in the EMBL-EBI ArrayExpress repository under number E-MTAB-9044.

2021-01-01 | E-MTAB-9807 | biostudies-arrayexpress

MicroRNA profiling of human cholangiocyte H69 cells after culturing with lipopolysaccharide

Project description:H69 cells were cultured in H69 medium with 1 ng/ml lipopolysaccharide(LPS, for smaples 04, 05 and 06) or without LPS(for samples 01, 02 and 03) for 8 hours and then collected for array analysis. <br>

2010-02-11 | E-MEXP-2052 | biostudies-arrayexpress

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