FGFR2 fusion proteins drive oncogenic transformation of mouse liver organoids towards cholangiocarcinoma

Abstract Understanding the mechanism of how cholangiocytes (liver ductal cells) are activated upon liver injury and specified to hepatocytes would permit liver regenerative medicine. Here we achieved long-term in vitro expansion of mouse liver organoids by modulating signaling pathways with a combination of three small-molecule compounds. CHIR-99021, blebbistatin, and forskolin together maintained the liver organoids in bipotential stage with both cholangiocyte- and hepatocyte-specific gene expression profiles and enhanced capacity for further hepatocyte differentiation. By employing a chemical approach, we demonstrated that Wnt/β-catenin, NMII–Rac, and PKA–ERK are core signaling pathways essential and sufficient for mouse liver progenitor expansion. Moreover, the advanced small-molecule culture of bipotential organoids facilitates the ex vivo investigation of liver cell fate determination and the application of organoids in liver regenerative medicine.

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Abstract 2534: High-affinity small molecule inhibitors of the menin-MLL interaction reverse oncogenic transformation mediated by MLL fusion proteins in leukemia

10.1158/1538-7445.am2014-2534 ◽

2014 ◽

Author(s):

Jolanta E. Grembecka ◽

Shihan He ◽

Timothy J. Senter ◽

Dmitry Borkin ◽

Jonathan Pollock ◽

...

Keyword(s):

Small Molecule ◽

Fusion Proteins ◽

Small Molecule Inhibitors ◽

Oncogenic Transformation ◽

High Affinity

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Trimethylamine n-Oxide (TMAO) Modulates the Expression of Cardiovascular Disease-Related microRNAs and Their Targets

International Journal of Molecular Sciences ◽

10.3390/ijms222011145 ◽

2021 ◽

Vol 22 (20) ◽

pp. 11145

Author(s):

Laura Díez-Ricote ◽

Paloma Ruiz-Valderrey ◽

Víctor Micó ◽

Ruth Blanco-Rojo ◽

João Tomé Carneiro ◽

...

Keyword(s):

Mouse Liver ◽

Target Gene ◽

Target Genes ◽

Molecular Pathways ◽

Cvd Risk ◽

Animal Products ◽

Human Macrophages ◽

Cellular Models ◽

Mice Liver ◽

Liver Organoids

Diet is a well-known risk factor of cardiovascular diseases (CVDs). Some microRNAs (miRNAs) have been described to regulate molecular pathways related to CVDs. Diet can modulate miRNAs and their target genes. Choline, betaine, and l-carnitine, nutrients found in animal products, are metabolized into trimethylamine n-oxide (TMAO), which has been associated with CVD risk. The aim of this study was to investigate TMAO regulation of CVD-related miRNAs and their target genes in cellular models of liver and macrophages. We treated HEPG-2, THP-1, mouse liver organoids, and primary human macrophages with 6 µM TMAO at different timepoints (4, 8, and 24 h for HEPG-2 and mouse liver organoids, 12 and 24 h for THP-1, and 12 h for primary human macrophages) and analyzed the expression of a selected panel of CVD-related miRNAs and their target genes and proteins by real-time PCR and Western blot, respectively. HEPG-2 cells were transfected with anti-miR-30c and syn-miR-30c. TMAO increased the expression of miR-21-5p and miR-30c-5p. PER2, a target gene of both, decreased its expression with TMAO in HEPG-2 and mice liver organoids but increased its mRNA expression with syn-miR-30c. We concluded that TMAO modulates the expression of miRNAs related to CVDs, and that such modulation affects their target genes.

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Ultrastructural localization of specific nucleoprotein fractions

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081887 ◽

1978 ◽

Vol 36 (2) ◽

pp. 204-205

Author(s):

G. L. Brown

Keyword(s):

Gel Electrophoresis ◽

Mouse Liver ◽

Polyacrylamide Gel Electrophoresis ◽

Ultrastructural Localization ◽

Polyacrylamide Gel ◽

Acid Extraction ◽

Acid Solutions ◽

Low Salt ◽

Liver Nuclei ◽

Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

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Site of Lysosome Production During Hepatic Injury

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100063652 ◽

1969 ◽

Vol 27 ◽

pp. 348-349

Author(s):

Nalin J. Unakar

Keyword(s):

Electron Microscopy ◽

Golgi Apparatus ◽

Mouse Liver ◽

Hepatic Injury ◽

Close Approximation ◽

Experimental Conditions ◽

Toxic Injury

The increased number of lysosomes as well as the close approximation of lysosomes to the Golgi apparatus in tissue under variety of experimental conditions is commonly observed. These observations suggest Golgi involvement in lysosomal production. The role of the Golgi apparatus in the production of lysosomes in mouse liver was studied by electron microscopy of liver following toxic injury by CCI4.

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Nuclear coiled bodies and the nucleolus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100167834 ◽

1994 ◽

Vol 52 ◽

pp. 20-21

Author(s):

K. Brasch ◽

J. Williams ◽

D. Gallo ◽

T. Lee ◽

R. L. Ochs

Keyword(s):

Mouse Liver ◽

Nuclear Body ◽

Nuclear Bodies ◽

Model Systems ◽

Coiled Body ◽

Rrna Processing ◽

Malignant Cells ◽

Sm Proteins ◽

Coiled Bodies ◽

Unique Protein

Though first described in 1903 by Ramon-y-Cajal as silver-staining “accessory bodies” to nucleoli, nuclear bodies were subsequently rediscovered by electron microscopy about 30 years ago. Nuclear bodies are ubiquitous, but seem most abundant in hyperactive and malignant cells. The best studied type of nuclear body is the coiled body (CB), so termed due to characteristic morphology and content of a unique protein, p80-coilin (Fig.1). While no specific functions have as yet been assigned to CBs, they contain spliceosome snRNAs and proteins, and also the nucleolar protein fibrillarin. In addition, there is mounting evidence that CBs arise from or are generated near the nucleolus and then migrate into the nucleoplasm. This suggests that as yet undefined links may exist, between nucleolar pre-rRNA processing events and the spliceosome-associated Sm proteins in CBs.We are examining CB and nucleolar changes in three diverse model systems: (1) estrogen stimulated chick liver, (2) normal and neoplastic cells, and (3) polyploid mouse liver.

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