• cytokinesis
• actin
• escrt
• microtubules

• Humans
• Actins/metabolism
• HeLa Cells
• Spastin/metabolism
• Microtubules/metabolism
• Cytokinesis
• Endosomal Sorting Complexes Required for Transport/metabolism
• Actin Depolymerizing Factors/metabolism

• Institut Pasteur
• Centre National de la Recherche Scientifique (National Center for Scientific Research)
• Agence Nationale de la Recherche (French National Research Agency)
• Fondation pour la Recherche Médicale (Foundation for Medical Research in France)

• Jag1
• Notch
• bile duct paucity
• biliary epithelial cells (BECs)
• cholangiocytes

• Mice
• Animals
• Alagille Syndrome/genetics
• Bile Ducts
• Biliary Tract
• Bile Ducts, Intrahepatic
• Organoids/metabolism

• Horizon Europe Marie Skłodowska-Curie Actions
• Karolinska Institutet
• Ragnar Söderberg Foundation
• Swedish Foundations Starting Grant
• University of the Punjab
• Vetenskapsrådet
• Wera Ekströms Stiftelse för Pediatrisk Forskning

• liver
• regeneration
• biliary epithelial cell
• notch
• yap

• Animals
• Zebrafish
• Cell Transdifferentiation/physiology
• Hepatocytes/metabolism
• Liver
• Epithelial Cells
• Cholestasis/metabolism
• Liver Diseases/metabolism
• Cell Proliferation
• Liver Regeneration/physiology

• P30 DK120531 NIDDK NIH HHS
• R01 DK101426 NIDDK NIH HHS
• R01 DK116993 NIDDK NIH HHS
• R01 DK132014 NIDDK NIH HHS

• hepatocellular carcinoma
• immunotherapy
• targeted therapy
• tumor microenvironment

• Chronic Liver Injury
• LPLC
• Liver Regeneration

• Humans
• Liver Regeneration
• Interferon-gamma
• Liver Diseases
• Killer Cells, Natural
• Macrophages

• NICD
• Notch pathway
• cancer stem cells
• dysregulation
• γ secretase

To investigate whether the Notch–Hif-1α signaling pathway is involved in liver regeneration.

Rats were divided into two groups and treated with daily intraperitoneal injections of saline (control) or the gamma-secretase inhibitor, Fli-06, for 2 days. Two-thirds of the rat livers were resected and rats were later euthanized at specific time points post-resection to analyze the remnant livers. Each group's liver/body weight ratio was calculated, and immunostaining and western blotting were used to determine the cell proliferation marker, PCNA and Ki-67 expression. Real-time PCR and western blotting were used to compare the mRNA expression of Notch homolog-1 (Notch1), hairy and enhancer of split-1 (Hes1), and vascular endothelial growth factor (Vegf), and the protein expression of NICD and HIF-1α, respectively.

The liver/body weight ratios and number of Ki-67- and PCNA-positive cells were significantly lower in the experimental group than the control group, indicating lower levels of liver regeneration following the disruption of Notch signaling by Fli-06. The Hes1 and Vegf mRNA levels and NICD and HIF-1α protein expression levels were all down-regulated by Fli-06 treatment.

Notch–Hif-α signaling pathway activation plays an important role in liver regeneration, where it may contribute toward liver cell proliferation.

• Hif-1α
• Liver regeneration
• Notch homolog-1
• Notch intracellular domain
• gamma-secretase inhibitors
• hepatectomy
• vascular endothelial growth factor

• Cytokines
• Hepatic progenitor cells
• Liver regeneration
• Micro ribonucleic acid
• Molecular pathways
• Partial hepatectomy

As a highly conserved signaling pathway in metazoans, the Notch pathway plays important roles in embryonic development and tissue regeneration. Recently, cardiac injury and regeneration have become an increasingly popular topic for biomedical research, and Notch signaling has been shown to exert crucial functions during heart regeneration as well. In this review, we briefly summarize the molecular functions of the endocardial Notch pathway in several cardiac injury and stress models. Although there is an increase in appreciating the importance of endocardial Notch signaling in heart regeneration, the mechanism of its activation is not fully understood. This review highlights recent findings on the activation of the endocardial Notch pathway by hemodynamic blood flow change in larval zebrafish ventricle after partial ablation, a process involving primary cilia, mechanosensitive ion channel Trpv4 and mechanosensitive transcription factor Klf2.

• Heart regeneration
• Hemodynamic alteration
• Ion channel
• Notch signaling
• Primary cilium
• klf2

• Cholangiocarcinoma
• Covalently closed circular DNA
• Hepatitis B virus
• Hepatocellular carcinoma
• Liver cancer
• Liver cancer stem cells
• Notch signaling
• α-Fetoprotein

• inflammation
• liver cancer
• microenvironment
• periportal
• progenitor cell

• Leydig cell
• P21Waf1/Cip1
• P27
• cell proliferation

• Animals
• Benzene Derivatives/pharmacology
• Cell Line
• Cell Proliferation/drug effects
• Cyclin-Dependent Kinase Inhibitor p21/metabolism
• Cyclin-Dependent Kinase Inhibitor p27/metabolism
• G1 Phase Cell Cycle Checkpoints/drug effects
• Leydig Cells/drug effects
• Leydig Cells/physiology
• Male
• Mice
• Propionates/pharmacology
• Receptors, Notch/antagonists & inhibitors
• Receptors, Notch/metabolism
• Signal Transduction/drug effects
• Signal Transduction/physiology
• Sulfones/pharmacology

• 81601242 National Natural Science Foundation of China
• 81660332 National Natural Science Foundation of China

The Notch signaling pathway, known to be a highly conserved signaling pathway in embryonic development and adult tissue homeostasis, participates in cell fate decisions that include cellular differentiation, cell survival and cell death. However, other studies have shown that aberrant in Notch signaling is pro-tumorigenic, particularly in hepatocellular carcinoma (HCC). HCC is one of the most common malignant tumors in the world and has a high mortality rate. Growing evidence supports that Notch signaling plays a critical role in the development of HCC by regulating the tumor microenvironment, tumorigenesis, progression, angiogenesis, invasion and metastasis. Accordingly, overexpression of Notch is closely associated with poor prognosis in HCC. In this review, we focus on the pro-tumorigenic role of Notch signaling in HCC, summarize the current knowledge of Notch signaling and its role in HCC development, and outline the therapeutic potential of targeting Notch signaling in HCC.

• Notch signaling pathway
• carcinogenesis
• hepatocellular carcinoma
• therapeutics

The immunologic interaction between parenchyma cells and encircling inflammatory cells is thought to be the most important mechanism of biliary damage and repair in primary sclerosing cholangitis (PSC). Monocytes/macrophages as master regulators of hepatic inflammation have been demonstrated to contribute to PSC pathogenesis. Macrophages coordinate with liver regeneration, and multiple phenotypes have been identified with diverse expressions of surface proteins and cytokine productions. We analyzed the expression of Notch ligand Jagged1 in polarized macrophages and investigated the relevance of Notch signalling activation in liver regeneration. M1 or M2 macrophages were generated from mouse bone marrow-derived macrophages (BMDMs) by classical or alternative activation, respectively. Then, the expression levels of Jagged1 (Jag1) of each phenotype were measured. The effects of polarized BMDMs on the expression of hepatic progenitor cell- (HPC-) specific markers and hairy and enhancer of split-1 (HES1) in HPCs in coculture were also analyzed. Monocyte-macrophage and Notch signalling-associated gene signatures were evaluated in the GEO database (access ID: GSE61260) by gene set enrichment analysis (GSEA). M1 macrophages were found associated with elevated Jag1 expression, which increased the fraction of HPC with self-renewing phenotypes (CD326⁺CD44⁺ or CD324⁺CD44⁺) and HES1 expression level in cocultured HPC. Blocking Jagged1 by siRNA or antibody in the coculture system attenuates HPC self-renewing phenotypes as well as HES1 expression in HPC. GSEA data show that macrophage activation and Notch signalling-associated gene signatures are enriched in PSC patients. These findings suggest that M1 macrophages promote an HPC self-renewing phenotype which is associated with Notch signalling activation within HPC. In the liver of PSC patients, the prevalence of activated macrophages, with M1 polarized accounting for the main part, is associated with increment of Notch signalling and enhancement of HPC self-renewal.

During embryonic development bipotential hepatoblasts differentiate into hepatocytes and cholangiocytes- the two main cell types within the liver. Cell fate decision depends on elaborate interactions between distinct signalling pathways, namely Notch, WNT, TGFβ, and Hedgehog. Several in vitro protocols have been established to differentiate human pluripotent stem cells into either hepatocyte or cholangiocyte like cells (HLC/CLC) to enable disease modelling or drug screening. During HLC differentiation we observed the occurrence of epithelial cells with a phenotype divergent from the typical hepatic polygonal shape- we refer to these as endoderm derived epithelial cells (EDECs). These cells do not express the mature hepatocyte marker ALB or the progenitor marker AFP. However they express the cholangiocyte markers SOX9, OPN, CFTR as well as HNF4α, CK18 and CK19. Interestingly, they express both E Cadherin and Vimentin, two markers that are mutually exclusive, except for cancer cells. EDECs grow spontaneously under low density cell culture conditions and their occurrence was unaffected by interfering with the above mentioned signalling pathways.

• Cadherins/metabolism
• Cell Count
• Cell Culture Techniques
• Cell Differentiation/physiology
• Cell Line
• Epithelial Cells/cytology
• Epithelial Cells/physiology
• Hepatocytes/cytology
• Hepatocytes/physiology
• Humans
• Induced Pluripotent Stem Cells/cytology
• Induced Pluripotent Stem Cells/physiology
• Transcriptome
• Vimentin/metabolism

• Medizinische Fakultät, Heinrich-Heine-Universität Düsseldorf

• Notch transcription complex
• blocking antibody
• clinical trials
• combination therapy
• gamma secretase inhibitors

Hepatocyte nuclear factor-1beta plays an important role in the development and progression of liver cancer. In recent years, the expression of HNF-1β has been reported to be associated with risk for a variety of cancers. The purpose of this study is to investigate whether the expression of HNF-1β promotes the malignancy of HCC and its mechanism. We retrospectively investigated the expression of HNF-1β in 90 patients with hepatocellular carcinoma and found that the high expression of HNF-1β indicated poor prognosis. We overexpressed HNF-1β in liver cancer cell lines and found the expression of liver progenitor cell markers and stemness were upregulated. The invasion ability and epithelial-mesenchymal transition (EMT)-associated genes were also significantly higher in liver cancer cells overexpressing HNF-1β than in the control group. A mechanistic study suggested the activation of the Notch signalling pathway probably plays a key role downstream of HNF-1β. More importantly, HNF-1β promoted tumourigenesis of HCC cells in vivo. In conclusion, high expression of HNF-1β not only promoted the de-differentiation of HCC cells into liver cancer stem cells through activating the Notch pathway but also enhanced the invasive potential of HCC cells and EMT occurrence, which would contribute to the enhancement of cell migration and invasion.

• Development
• Luminal cell differentiation
• MYC
• NOTCH
• Prostate
• p38-MAPK

The Notch signaling pathway is a very conserved system that controls embryonic cell fate decisions and the maintenance of adult stem cells through cell to cell communication. Accumulating evidence support the relevance of Notch signaling in different human diseases and it is one of the most commonly activated signaling pathways in cancer. This review focuses mainly on the role of Notch3 signaling in hepatocellular carcinoma and its potential therapeutic applications against this malignancy. In this regard, the crosstalk between Notch and p53 may play an important role.

• Notch
• hepatocellular carcinoma (HCC)
• p53

• Cirrhosis
• DNA repair
• Hepatic stellate cells
• Insulin-like growth factor I
• Liver macrophages
• MSCs

• Animals
• Cells, Cultured
• Down-Regulation/drug effects
• Gene Expression/drug effects
• Hepatic Stellate Cells/drug effects
• Hepatic Stellate Cells/metabolism
• Hepatic Stellate Cells/physiology
• Hepatocyte Growth Factor/metabolism
• Hepatocytes/drug effects
• Hepatocytes/metabolism
• Insulin-Like Growth Factor I/metabolism
• Liver/drug effects
• Liver/metabolism
• Liver Cirrhosis/metabolism
• Liver Cirrhosis/therapy
• Macrophages/cytology
• Macrophages/drug effects
• Macrophages/metabolism
• Male
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Mice
• Mice, Inbred BALB C
• Thioacetamide/pharmacology
• Up-Regulation/drug effects

• ANPCyT
• CONICET

• Notch
• bile duct
• cancer
• cholangiocarcinoma
• noncanonical

• Animals
• Carcinogenesis/genetics
• Cholangiocarcinoma/genetics
• Cholangiocarcinoma/pathology
• Humans
• Immunoglobulin Joining Region/genetics
• Mice
• Mice, Transgenic
• Neoplasms, Experimental/genetics
• Neoplasms, Experimental/pathology
• Phosphatidylinositol 3-Kinases/genetics
• Prognosis
• Rats
• Receptor, Notch3/genetics
• Signal Transduction
• Tumor Suppressor Protein p53/genetics

• 21139 Cancer Research UK
• 14889 Cancer Research UK
• 12946 Cancer Research UK
• MR/K017047/1 Medical Research Council
• 20409 Cancer Research UK
• 11650 Cancer Research UK
• A11650 Cancer Research UK
• Wellcome Trust

• Breast cancer
• Degradation
• Notch
• Therapy
• Ubiquitin-proteasome system

• Animals
• Breast Neoplasms/drug therapy
• Breast Neoplasms/metabolism
• Breast Neoplasms/pathology
• Humans
• Ligands
• Receptors, Notch/metabolism
• Signal Transduction
• Ubiquitination

The bipotential differentiation of liver progenitor cells underlies liver development and bile duct formation as well as liver regeneration and disease. TGFβ and Notch signaling are known to play important roles in the liver progenitor specification process and tissue morphogenesis. However, the complexity of these signaling pathways and their currently undefined interactions with other microenvironmental factors, including extracellular matrix (ECM), remain barriers to complete mechanistic understanding. Utilizing a series of strategies, including co-cultures and cellular microarrays, we identified distinct contributions of different Notch ligands and ECM proteins in the fate decisions of bipotential mouse embryonic liver (BMEL) progenitor cells. In particular, we demonstrated a cooperative influence of Jagged-1 and TGFβ1 on cholangiocytic differentiation. We established ECM-specific effects using cellular microarrays consisting of 32 distinct combinations of collagen I, collagen III, collagen IV, fibronectin, and laminin. In addition, we demonstrated that exogenous Jagged-1, Delta-like 1, and Delta-like 4 within the cellular microarray format was sufficient for enhancing cholangiocytic differentiation. Further, by combining Notch ligand microarrays with shRNA-based knockdown of Notch ligands, we systematically examined the effects of both cell-extrinsic and cell-intrinsic ligand. Our results highlight the importance of divergent Notch ligand function and combinatorial microenvironmental regulation in liver progenitor fate specification.

Aberrant transcriptional regulation contributes to the pathogenesis of both congenital and adult forms of liver disease. Although the transcription factor RBPJ is essential for liver morphogenesis and biliary development, its specific function in the differentiation of hepatic progenitor cells (HPC) has not been investigated, and little is known about its role in adult liver regeneration. HPCs are bipotent liver stem cells that can self-replicate and differentiate into hepatocytes or cholangiocytes in vitro. HPCs are thought to play an important role in liver regeneration and repair responses. While the coordinated repopulation of both hepatocyte and cholangiocyte compartment is pivotal to the structure and function of the liver after regeneration, the mechanisms coordinating biliary regeneration remain vastly understudied. Here, we utilized complex genetic manipulations to drive liver-specific deletion of the Rbpj gene in conjunction with lineage tracing techniques to delineate the precise functions of RBPJ during biliary development and HPC-associated biliary regeneration after hepatectomy. Furthermore, we demonstrate that RBPJ promotes HPC differentiation toward cholangiocytes in vitro and blocks hepatocyte differentiation through mechanisms involving Hippo-Notch crosstalk. Overall, this study demonstrates that the Notch-RBPJ signaling axis critically regulates biliary regeneration by coordinating the fate decision of HPC and clarifies the molecular mechanisms involved.

Activation of the hypoxia-inducible factor (HIF)-pathway in hepatocellular carcinoma (HCC) induces therapy resistant tumours, characterized by increased liver progenitor cell (LPCs) characteristics and poor prognosis. We previously reported corresponding results in mice with HCC in which hypoxia was mimicked by prolyl hydroxylase domain (PHD) inhibition. Here, we aimed at investigating whether induction of LPC characteristics occurs during the onset of hepatocarcinogenesis and if this is associated with activation of Notch signalling. Dietheylnitrosamine (DEN) was used to induce hepatic tumours in PHD2 haplodeficient (PHD2^+/-) mice which were euthanized at 5, 10, 15 and 17 weeks following DEN during neoplastic transformation, before tumour formation. Neoplasia and mRNA expression of LPC and Notch markers were evaluated by histology and qPCR on isolated livers. PHD2 haplodeficiency resulted in enhanced expression of HIF target genes after 17 weeks of DEN compared to wild type (WT) littermates but had no effect on the onset of neoplastic transformation. The mRNA expression of Afp and Epcam was increased at all time points following DEN whereas CK19, Prom1 and Notch3 were increased after 17 weeks of DEN, without difference between PHD2^+/- and WT mice. MDR1 mRNA expression was increased in all DEN treated mice compared to saline control with increased expression in PHD2^+/- compared to WT from 15 weeks. These results indicate that the effects of PHD2 haplodeficiency on the expression of LPC and Notch markers manifest during tumour nodule formation and not early on during neoplastic transformation.

• diethylnitrosamine
• hepatocarcinogenesis
• hypoxia
• liver progenitor cells
• notch
• prolyl hydroxylase domain

• Cholangiocarcinoma (CC)
• Hepatoblastoma (HBT)
• Hepatocellular carcinoma (HCC)
• Liver cancer
• NOTCH ligand
• NOTCH receptor

• Human embryonic stem cell
• Induced pluripotent stem cell
• Krüppel-like factor 9
• Notch
• Thyroid receptor

• Animals
• Cell Differentiation/genetics
• Cell Transformation, Neoplastic/genetics
• Cell Transformation, Neoplastic/metabolism
• Drosophila melanogaster/genetics
• Humans
• Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism
• Neoplasms/drug therapy
• Neoplasms/metabolism
• Receptors, Notch/antagonists & inhibitors
• Receptors, Notch/genetics
• Receptors, Notch/metabolism
• Signal Transduction/genetics

• R01 CA173636 NCI NIH HHS
• 1R01CA172560 NCI NIH HHS
• R01 CA133379 NCI NIH HHS
• R01 CA149655 NCI NIH HHS
• R01 CA105129 NCI NIH HHS
• R01 CA114102 NCI NIH HHS
• R01 CA169784 NCI NIH HHS
• Howard Hughes Medical Institute
• R01 CA172560 NCI NIH HHS