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Single cell RNA-seq of Sox9-deficient intrahepatic biliary epithelial cells

ABSTRACT: To define the regulatory role of Sox9 in intrahepatic biliary epithelium, we conducted single cell RNA-seq on biliary epithelial cells (BECs) from adult control (Sox9fl/fl) and conditional Sox9 knockout (Sox9cKO; Sox9fl/fl:Albumin-Cre) mice. Livers were dissociated to single cells and BECs isolated by FACS based on CD326 positivity. CD31+ and CD45+ cells were excluded by FACS. Gallbladder and extrahepatic tissues were excluded by dissection. Sorted cells were fixed according to manufacturer instructions using Parse Whole Cell Fixation kit. Fixed cells were frozen and stored at -70C for less than 6mo, prior to being used in split-pool combinatorial barcoding according to manufacturer instructions. Barcoding and library construction was carried out with the Parse Evercode Whole Transcriptome v2 kit.

ORGANISM(S): Mus musculus

PROVIDER: GSE249558 | GEO | 2024/01/01

REPOSITORIES: GEO

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Bulk RNA-seq of Sox9-deficient intrahepatic biliary epithelial cells

Project description:To define the regulatory role of Sox9 in intrahepatic biliary epithelium, we conducted bulk RNA-seq on biliary epithelial cells (BECs) from adult control (Sox9fl/fl) and conditional Sox9 knockout (Sox9cKO; Sox9fl/fl:Albumin-Cre) mice. Livers were dissociated to single cells and BECs isolated by FACS based on CD326 positivity. CD31+ and CD45+ cells were excluded by FACS. Gallbladder and extrahepatic tissues were excluded by dissection. RNA was isolated using Ambion RNAqueous Micro kit and libraries were made using Takara SMARTer Stranded Total RNA-seq Kit v3 Pico.

2024-01-01 | GSE249385 | GEO

RNA-seq of intrahepatic Sox9EGFP populations

Project description:Cellular heterogeneity in the biliary epithelium is poorly understood, and genetic biomarkers for identifying distinct biliary epithelial cell (BEC) subpopulations are limited. Here, we utilized a Sox9EGFP transgene to study cellular heterogeneity in intrahepatic BECs. Male Sox9EGFP mice were fasted overnight, and livers were dissociated to obtain single cells. Gallbladders and extrahepatic ducts were excluded by dissection. Sox9EGFP populations were isolated by FACS into four distinct populations: GFPnegative, GFPsublow, GFPlow, and GFPhigh. We conducted RNA-seq and differential gene expression analysis to identify genes shared between and unique to each population.

2020-05-29 | GSE151387 | GEO

Downregulation of TGR5 (GPBAR1) in biliary epithelial cells is characteristic of sclerosing cholangitis and may contribute to disease pathogenesis

Project description:Primary sclerosing cholangitis (PSC) is characterized by chronic inflammation and progressive fibrosis of the biliary tree. The bile acid receptor TGR5 is found on biliary epithelial cells (BECs), where it promotes secretion, proliferation and tight junction integrity. Thus, we speculated that changes in TGR5 expression in BECs may contribute to PSC pathogenesis.

2021-03-12 | GSE168758 | GEO

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Project description:We report transcriptional profiles of liver biliary epithelial cells (BECs) from non-pregnant mice and pregnant mice. By RNA-sequencing of FACS-isolated BECs, we revealed that genes related to tissue morphogenesis and actin cytoskeleton were upregulated on dpc8.

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Dynamic epigenetic and transcriptional changes drive cellular plasticity in the liver

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Transcriptional profiling of hepatocyte, reprogrammed hepatocyte, and biliary cells

Project description:Following liver injury, a subset of hepatocytes adopt features of biliary epithelial cells (BECs) in a process known as biliary reprogramming. The aim of this study is to elucidate the molecular events accompanying this dramatic shift in cellular identity. Wild type hepatocytes, wild type biliary cells, DDC-treated hepatocytes, DDC-treated biliary cells, and DDC-treated reprogrammed cells were isolated by FACS, RNA isolated, and sequenced.

2020-08-27 | GSE156894 | GEO

VEGFA mRNA-LNP promote biliary epithelial cell-to-hepatocyte conversion in acute and chronic liver diseases and reverses steatosis and fibrosis

Project description:The goal of this experiment was to analyse transcriptomic changes between Biliary epithelial cells or BECs from the liver of mice treated with Acetaminophen (a drug that causes acute liver injury) in combination with AAV8-Tbg-p21 viral vector to induce hepatocyte senecence. The aim was to understand the impact of injury on transcriptional status of biliary epithelial cells. Briefly, we analyzed mouse liver cells (hepatocytes and non-parenchymal cells fractions) obtained by in-situ liver perfusion 48 h after injury. We further extracted cells corresponding to the hepatocyte and biliary epithelial cell gene signature for detailed analysis. Two separate clusters of BECs (BEC1 and BEC2) were further identified and analyzed to study their transcriptional changes and corresponding pathways.

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Identification of Sox9-Regulated Pathways During Early Pancreas Organogenesis

Project description:Sox9 target genes were identified by comparing gene expression in Sox9-ablated versus wild-type pancreata using microarray analysis. Sox9 was conditionally ablated in the developing pancreas via recombination of a Sox9-flox allele (Kist et al., 2002) using the Pdx1-Cre transgene (Gu et al., 2002). Total RNA was isolated and pooled from dorsal pancreatic epithelia of e12.5 Sox9fl/fl; Pdx1-Cre (mutant) versus Sox9fl/fl (wild-type) littermates for three biological replicates.

2012-08-01 | E-GEOD-28669 | biostudies-arrayexpress

Transcriptional changes of adult liver biliary epithelial cells in vivo upon high-fat diet

Project description:Background and Aims: During severe or chronic hepatic injury, biliary epithelial cells (BECs), also known as cholangiocytes, undergo rapid reprogramming and proliferation, a process known as ductular reaction (DR), and allow liver regeneration by differentiating into both functional cholangiocytes and hepatocytes. While DR is a hallmark of chronic liver diseases, including advanced stages of non-alcoholic fatty liver disease (NAFLD), the early events underlying BEC activation are largely unknown. Since NAFLD initiates with increased lipid accumulation, a stage called steatosis; we hypothesized that BECs isolated directly from steatotic livers might address current knowledge gaps in early BEC activation. Approach and Results: Here, we used methods allowing isolation and extensive transcriptional characterization of BECs from chow diet (CD)- and high-fat diet (HFD)-fed mice livers. We demonstrate that BECs readily accumulate lipids during HFD feeding and that lipid overload induces the conversion of adult cholangiocytes into active BECs through upregulation of the cell cycle and DNA replication signature. This event is accompanied by significant downregulation of extracellular matrix organization in BECs derived from HFD-fed mice livers but not CD-fed mice livers. Mechanistically, we found that lipid overload unleashes the activation of the E2F transcription factors in BECs, which drives cell cycle progression.

2023-03-22 | GSE217739 | GEO

Profiling of Sox9-related genes in chondrogenesis using Sox9-3'EGFP knock-in and Sox9-EGFP/EGFP null chimeric mouse embryos

Project description:To define the repertoire of Sox9-dependent genes that contribute to the regulation of chondrogenesis, we generated Sox9-3'enhanced green fluorescent protein (EGFP) knock-in mice (Sox9-3'EGFP) and Sox9-EGFP/EGFP null chimeras. EGFP-positive cells of Sox9-3'EGFP knock-in and Sox9-EGFP/EGFP null chimeric embryos harvested from limb buds at embryonic day 12.5 were sorted using a FACSAria flow cytometer (Becton-Dickinson). Total RNA of sorted cells was extracted using the RNeasy Mini Kit (QIAGEN) and amplified according to the instructions provided by Affymetrix. Microarray analysis using the Affymetrix Mouse Genome 430 2.0 Array was performed according to the manufacturer's instructions.

2011-02-16 | E-MTAB-537 | biostudies-arrayexpress

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