Clec3b⁺ fibroblasts are the primary effectors of portal fibrosis following activation via a KLF4/periostin axis
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE298824 | GEO | 2026/03/16
REPOSITORIES: GEO
Similar Datasets
Project description:Periportal fibrosis is a hallmark of cholestatic liver injury, yet the cellular origins and regulatory mechanisms underlying periportal fibrogenesis remain unclear. This study aims to identify the cellular origins and master regulators of cholestatic periportal fibrosis. Single-cell RNA sequencing was performed on bile duct-ligated (BDL) mouse model. A novel Clec3b-tdTomato lineage-tracing mouse model (Clec3bCreERT2) was generated to track portal fibroblast myofibroblastic activation in both BDL and 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) mouse models. Conditional deletion of Klf4 with Clec3bCreERT2 and adeno-associated virus (AAV6)-mediated KLF4 overexpression were employed to investigate the function of KLF4 in cholestatic liver diseases models. RNA sequencing (RNA-seq) and chromatin immunoprecipitation-sequencing (ChIP-seq) were used to identify KLF4 binding targets. AAV6 carrying short hairpin RNA targeting periostin (POSTN) and neutralizing antibodies were used to gain insight into the underlying mechanisms of POSTN. Single-cell analysis identified MFAP4 as a specific marker of portal fibroblasts and their myofibroblast derivatives during cholestatic liver fibrosis. Cell-fate mapping demonstrated that Clec3b+ portal fibroblasts serve as progenitors of myofibroblasts, and contribute to periportal fibrosis. KLF4 was identified as a pivotal regulator of myofibroblastic activation in portal fibroblasts, with expression inversely correlated with fibrosis severity. Mechanistically, KLF4 represses POSTN expression by binding to its promoter, while POSTN, through interaction with integrin αvβ5, activates fibrogenic signaling in Clec3b+ portal fibroblasts. This study provides new insights into the cellular and molecular mechanisms governing periportal fibrosis in cholestatic liver disease. We identify the Clec3b+ portal fibroblasts as critical contributors to periportal fibrosis and highlight the KLF4/POSTN signaling axis as a key regulatory pathway, offering potential therapeutic approaches for cholestatic liver fibrosis.
2026-03-16 | GSE298651 | GEO
Project description:Periportal fibrosis is a hallmark of cholestatic liver injury, yet the cellular origins and regulatory mechanisms underlying periportal fibrogenesis remain unclear. This study aims to identify the cellular origins and master regulators of cholestatic periportal fibrosis. Single-cell RNA sequencing was performed on bile duct-ligated (BDL) mouse model. A novel Clec3b-tdTomato lineage-tracing mouse model (Clec3bCreERT2) was generated to track portal fibroblast myofibroblastic activation in both BDL and 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) mouse models. Conditional deletion of Klf4 with Clec3bCreERT2 and adeno-associated virus (AAV6)-mediated KLF4 overexpression were employed to investigate the function of KLF4 in cholestatic liver diseases models. RNA sequencing (RNA-seq) and chromatin immunoprecipitation-sequencing (ChIP-seq) were used to identify KLF4 binding targets. AAV6 carrying short hairpin RNA targeting periostin (POSTN) and neutralizing antibodies were used to gain insight into the underlying mechanisms of POSTN. Single-cell analysis identified MFAP4 as a specific marker of portal fibroblasts and their myofibroblast derivatives during cholestatic liver fibrosis. Cell-fate mapping demonstrated that Clec3b+ portal fibroblasts serve as progenitors of myofibroblasts, and contribute to periportal fibrosis. KLF4 was identified as a pivotal regulator of myofibroblastic activation in portal fibroblasts, with expression inversely correlated with fibrosis severity. Mechanistically, KLF4 represses POSTN expression by binding to its promoter, while POSTN, through interaction with integrin αvβ5, activates fibrogenic signaling in Clec3b+ portal fibroblasts. This study provides new insights into the cellular and molecular mechanisms governing periportal fibrosis in cholestatic liver disease. We identify the Clec3b+ portal fibroblasts as critical contributors to periportal fibrosis and highlight the KLF4/POSTN signaling axis as a key regulatory pathway, offering potential therapeutic approaches for cholestatic liver fibrosis.
2026-03-16 | GSE298633 | GEO
Project description:A hallmark of pulmonary fibrosis is aberrant activation of lung fibroblasts into pathological fibroblasts producing excessive extracellular matrix (ECM). Thus, identification of key regulator(s) driving the generation of pathological fibroblasts can inform effective countermeasures against disease progression. In this study, we show that Leptin Receptor (Lepr)+ fibroblasts arising during alveologenesis include Signal Peptide-CUB-EGF Domain-containing Protein 2 (Scube2)+ alveolar fibroblasts as a major constituent significantly contributing to collagen triple helix repeat containing 1 (Cthrc1)+ Periostin (Postn)+ pathological fibroblasts in two mouse models of pulmonary fibrosis. Genetic ablation of the Postn+ pathological fibroblasts attenuates fibrosis. Comprehensive analysis of single cell RNA sequencing (scRNA-seq) and single cell Assay for Transposase-Accessible Chromatin sequencing (scATAC-seq) reveal Runt related transcription factor 2 (RUNX2) as a key regulator of fibrotic genes. Consistently, conditional deletion of Runx2 with Lepr-CreERT2 or Scube2-CreERT2 reduces the generation of pathological fibroblasts, ECM deposition, and pulmonary fibrosis. Therefore, Lepr+ derived fibroblasts which include Scube2+ alveolar fibroblasts are a key source of pathological fibroblasts, and targeting Runx2 provides a potential treatment option for pulmonary fibrosis.
2024-11-25 | GSE278419 | GEO
Project description:There are a few markers available to distinguish hepatic stellate cells (HSCs), portal fibroblasts (PFs), and mesothelial cells (MCs) in the adult mouse liver. To identify genes uniquely expressed in these cells, we isolated HSCs having Vitamin A lipids by FACS based on their autofluorescence. We isolated a mixed population containing PFs and MCs as Vitamin A lipid-negative and GFP-positive cells by FACS from Collagen1a1 promoter-driven GFP transgenic mice. MCs were isolated by FACS using anti-GPM6A antibodies from adult mouse livers. PFs were isolated by FACS as Vitamin A lipid-negative, GFP-positive, and GPM6A-negative cells. We also isolated HSCs, MCs, and PFs from Collagen1a1 promoter-driven GFP transgenic mouse livers having biliary fibrosis by FACS. Biliary fibrosis was induced by bile duct ligation for 3 weeks. Sham operation was performed as a control. After separation of these cells, we isolated total RNAs and analyzed mRNA expression by microarray analysis. Total RNA was extracted with RNAqueous Micro (Ambion) and the probes for the microarray were synthesized using the Ovation RNA amplification system V2 and FL-Ovation cDNA Biotin module V2 (Nugen). The labeled probes were hybridized with GeneChip Mouse Genome 430 2.0 arrays (Affymetrix) and signals were analyzed with Genomic Suite software (Partek).
2016-03-11 | E-GEOD-66788 | biostudies-arrayexpress
Project description:Modelling liver disease requires in vitro systems that replicate disease progression1,2. Current tissue-derived organoids fail to reproduce the complex cellular composition and tissue architecture observed in vivo3. Here, we describe a multicellular organoid system composed of adult hepatocytes, cholangiocytes and mesenchymal cells that recapitulates the architecture of the liver periportal region and, when manipulated, models aspects of cholestatic injury and biliary fibrosis. We first generate reproducible hepatocyte organoids with functional bile canaliculi network that retain morphological features of in vivo tissue. By combining these with cholangiocytes and portal fibroblasts, we generate assembloids that mimic the cellular interactions of the periportal region. Assembloids are functional, consistently draining bile from bile canaliculi into the bile duct. Strikingly, manipulating the relative number of portal mesenchymal cells is sufficient to induce a fibrotic-like state, independently of an immune compartment. By generating chimeric assembloids of mutant and wild-type cells, or after gene knockdown, we show proof-of-concept that our system is amenable to investigating gene function and cell-autonomous mechanisms. Taken together, we demonstrate that liver assembloids represent a suitable in vitro system to study bile canaliculi formation, bile drainage, and how different cell types contribute to cholestatic disease and biliary fibrosis, in an all-in-one model.
2025-04-29 | GSE274973 | GEO
Project description:Modelling liver disease requires in vitro systems that replicate disease progression1,2. Current tissue-derived organoids fail to reproduce the complex cellular composition and tissue architecture observed in vivo3. Here, we describe a multicellular organoid system composed of adult hepatocytes, cholangiocytes and mesenchymal cells that recapitulates the architecture of the liver periportal region and, when manipulated, models aspects of cholestatic injury and biliary fibrosis. We first generate reproducible hepatocyte organoids with functional bile canaliculi network that retain morphological features of in vivo tissue. By combining these with cholangiocytes and portal fibroblasts, we generate assembloids that mimic the cellular interactions of the periportal region. Assembloids are functional, consistently draining bile from bile canaliculi into the bile duct. Strikingly, manipulating the relative number of portal mesenchymal cells is sufficient to induce a fibrotic-like state, independently of an immune compartment. By generating chimeric assembloids of mutant and wild-type cells, or after gene knockdown, we show proof-of-concept that our system is amenable to investigating gene function and cell-autonomous mechanisms. Taken together, we demonstrate that liver assembloids represent a suitable in vitro system to study bile canaliculi formation, bile drainage, and how different cell types contribute to cholestatic disease and biliary fibrosis, in an all-in-one model.
2025-04-29 | GSE274971 | GEO
Project description:Cardiac fibroblasts convert to myofibroblasts with injury to mediate healing after acute myocardial infarction and to mediate long-standing fibrosis with chronic disease. Myofibroblasts remain a poorly defined cell-type in terms of their origins and functional effects in vivo. Methods: Here we generate Postn (periostin) gene-targeted mice containing a tamoxifen inducible Cre for cellular lineage tracing analysis. This Postn allele identifies essentially all myofibroblasts within the heart and multiple other tissues. Results: Lineage tracing with 4 additional Cre-expressing mouse lines shows that periostin-expressing myofibroblasts in the heart derive from tissue-resident fibroblasts of the Tcf21 lineage, but not endothelial, immune/myeloid or smooth muscle cells. Deletion of periostin+ myofibroblasts reduces collagen production and scar formation after myocardial infarction. Periostin-traced myofibroblasts also revert back to a less activated state upon injury resolution. Conclusions: Our results define the myofibroblast as a periostin-expressing cell-type necessary for adaptive healing and fibrosis in the heart, which arises from Tcf21+ tissue-resident fibroblasts. Fluidigm C1 whole genome transcriptome analysis of lineage mapped cardiac myofibroblasts
2016-06-14 | E-GEOD-83337 | biostudies-arrayexpress
Project description:There are a few markers available to distinguish hepatic stellate cells (HSCs), portal fibroblasts (PFs), and mesothelial cells (MCs) in the adult mouse liver. To identify genes uniquely expressed in these cells, we isolated HSCs having Vitamin A lipids by FACS based on their autofluorescence. We isolated a mixed population containing PFs and MCs as Vitamin A lipid-negative and GFP-positive cells by FACS from Collagen1a1 promoter-driven GFP transgenic mice. MCs were isolated by FACS using anti-GPM6A antibodies from adult mouse livers. PFs were isolated by FACS as Vitamin A lipid-negative, GFP-positive, and GPM6A-negative cells. We also isolated HSCs, MCs, and PFs from Collagen1a1 promoter-driven GFP transgenic mouse livers having biliary fibrosis by FACS. Biliary fibrosis was induced by bile duct ligation for 3 weeks. Sham operation was performed as a control. After separation of these cells, we isolated total RNAs and analyzed mRNA expression by microarray analysis.
2016-03-11 | GSE66788 | GEO
Project description:Cardiac fibrosis is a common feature of ischemic heart disease and cardiac fibroblasts (CF) are key players in cardiac remodeling of the injured heart after myocardial infarction (MI). Fibrosis increases myocardial stiffness, thereby impairing cardiac function, which ultimately progresses to end-stage heart failure. Little is known, however, on the secretome of CF and cell-to-cell communication of CF is only incompletely understood. Here, we in vivo labeled secreted proteins by expressing TurboID under control of the POSTN promotor in cardiac fibroblasts of mouse with myocardial infarction, enriched biotinylated proteins and analyzed them using LC-MS.
2025-03-25 | PXD053791 | Pride
Project description:Biliary diseases can cause inflammation, fibrosis, bile duct destruction and eventually liver failure. There are no curative treatments for biliary disease except for liver transplantation. New therapies are urgently required. We have purified human Biliary Epithelial Cells (hBEC) from discarded human livers. hBECs were tested as a cell therapy in a mouse model of biliary disease where the conditional deletion of Mdm2 in cholangiocytes causes senescence, biliary strictures and fibrosis. hBEC are expandable, phenotypically stable and help restore biliary structure and function, highlighting their regenerative capacity and a potential alternative to liver transplantation for biliary disease.
2022-03-03 | GSE155498 | GEO