Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA281228 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:The routine study of human malaria liver-stage biology in vitro is hampered by low infection efficiency of human hepatocellular carcinoma (HCC) lines (<0.1%), poor understanding of steady-state HCC biology, and lack of appropriate tools for trace sample analysis. HC-04 is the only HCC that supports complete development of human malaria parasites. We hypothesized that HCCs are in various intermediate stages of the epithelial-mesenchymal transition (EMT) and HC-04s retain epithelial characteristics that permit infection. We developed an novel analytical approach to test this hypothesis viz. the HC-04 response to hepatocyte growth factor (HGF). We used online two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) to quantify protein expression profiles in HC-04 pre-/post-HGF treatment and validated these results by RT-qPCR and microscopy. We successfully increased protein identification efficiency over offline-2D methods by 12-fold using less sample material, allowing robust protein quantification. We observed expected up-regulation and down-regulation of EMT protein markers in response to HGF, but also unexpected cellular responses. HC-04 is thus susceptible to HGF-mediated signaling but to a lesser degree than what we observed for HepG2, a widely used, but poor malaria HCC model. Our analytical approach to understanding the basic biology of HC-04 helps us understand the factors that may influence its utility as a model for malaria liver-stage development. We observed that HC-04 treatment with HGF prior to the addition of Plasmodium falciparum sporozoites did not facilitate cell invasion, arguing for unlinking the effect of HGF on malaria liver stage development from hepatocyte invasion. Finally, our 2D-LC-MS/MS approach and broadly applicable experimental strategy should prove useful in the analysis of various hepatocyte-pathogen interactions, tumor progression and early disease events.
2016-07-20 | MSV000079959 | MassIVE
Project description:The routine study of human malaria liver-stage biology in vitro is hampered by low infection efficiency of human hepatocellular carcinoma (HCC) lines (<0.1%), poor understanding of steady-state HCC biology, and lack of appropriate tools for trace sample analysis. HC-04 is the only HCC that supports complete development of human malaria parasites. We hypothesized that HCCs are in various intermediate stages of the epithelial-mesenchymal transition (EMT) and HC-04s retain epithelial characteristics that permit infection. We developed an novel analytical approach to test this hypothesis viz. the HC-04 response to hepatocyte growth factor (HGF). We used online two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) to quantify protein expression profiles in HC-04 pre-/post-HGF treatment and validated these results by RT-qPCR and microscopy. We successfully increased protein identification efficiency over offline-2D methods by 12-fold using less sample material, allowing robust protein quantification. We observed expected up-regulation and down-regulation of EMT protein markers in response to HGF, but also unexpected cellular responses. HC-04 is thus susceptible to HGF-mediated signaling but to a lesser degree than what we observed for HepG2, a widely used, but poor malaria HCC model. Our analytical approach to understanding the basic biology of HC-04 helps us understand the factors that may influence its utility as a model for malaria liver-stage development. We observed that HC-04 treatment with HGF prior to the addition of Plasmodium falciparum sporozoites did not facilitate cell invasion, arguing for unlinking the effect of HGF on malaria liver stage development from hepatocyte invasion. Finally, our 2D-LC-MS/MS approach and broadly applicable experimental strategy should prove useful in the analysis of various hepatocyte-pathogen interactions, tumor progression and early disease events.
2014-02-17 | PXD000682 | Pride
Project description:In this study, we established human hepatocyte organoids (HHOs) that were expanded from primary human hepatocytes (PHH) in a defined medium. Briefly, the cryopreserved primary human hepatocytes (purchased) were thawn in advanced DMEM/F12 (Thermo) or Cryopreserved hepatocyte Recovery Medium (Gibco). Then, the cells were embedded in Matrigel (Corning) and cultured with the expansion medium (EM). For the eHHOs, expansion medium (EM) was prepared with a basal medium (Advanced DMEM/F12 was supplemented with penicillin/streptomycin, 10 mM HEPES, 2 mM GlutaMAX, 1 ×(Thermo Fisher Scientific), 10 nM gastrin I (Sigma), and 1 mM N-acetylcysteine (Wako, Japan) ) containing the following niche factors: 50 ng/ml mouse recombinant EGF (Thermo Fisher Scientific), 25 ng/mL human recombinant HGF (Peprotech), 100 ng/mL human recombinant FGF-10 (Peprotech), 10 uM Forskolin (Cayman Chemical), 25 ng/ml mouse recombinant noggin (Peprotech), 1 mg/ml human recombinant R-spondin1 (R; R&D), 20% Afamin-Wnt-3A serum-free conditioned medium (W; Mihara et al., 2016), 5 uM A83-01 (Tocris), and 20ng/ml human Oncostatin M (OSM; Peprotech). For dHHOs, eHHOs were cultured for 10-14 days in differentiation medium (DM), which was EM without OSM and WR, and containing a hormone cocktail (10ng/ml Growth Hormone, 10ng/ml Prolactin, and 100 ng/ml Cortisol) and 10uM DAPT (differentiation medium: DM).
2025-01-15 | GSE228249 | GEO
Project description:Mueller2015 - Hepatocyte proliferation, T160
phosphorylation of CDK2
This model is described in the article:
T160-phosphorylated CDK2
defines threshold for HGF-dependent proliferation in primary
hepatocytes.
Mueller S, Huard J, Waldow K, Huang
X, D'Alessandro LA, Bohl S, Börner K, Grimm D, Klamt S,
Klingmüller U, Schilling M.
Mol. Syst. Biol. 2015; 11(3): 795
Abstract:
Liver regeneration is a tightly controlled process mainly
achieved by proliferation of usually quiescent hepatocytes. The
specific molecular mechanisms ensuring cell division only in
response to proliferative signals such as hepatocyte growth
factor (HGF) are not fully understood. Here, we combined
quantitative time-resolved analysis of primary mouse hepatocyte
proliferation at the single cell and at the population level
with mathematical modeling. We showed that numerous G1/S
transition components are activated upon hepatocyte isolation
whereas DNA replication only occurs upon additional HGF
stimulation. In response to HGF, Cyclin:CDK complex formation
was increased, p21 rather than p27 was regulated, and Rb
expression was enhanced. Quantification of protein levels at
the restriction point showed an excess of CDK2 over CDK4 and
limiting amounts of the transcription factor E2F-1. Analysis
with our mathematical model revealed that T160 phosphorylation
of CDK2 correlated best with growth factor-dependent
proliferation, which we validated experimentally on both the
population and the single cell level. In conclusion, we
identified CDK2 phosphorylation as a gate-keeping mechanism to
maintain hepatocyte quiescence in the absence of HGF.
This model is hosted on
BioModels Database
and identified by:
BIOMD0000000568.
To cite BioModels Database, please use:
BioModels Database:
An enhanced, curated and annotated resource for published
quantitative kinetic models.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2024-09-02 | BIOMD0000000568 | BioModels
Project description:Two biological replicates of Madin-Darby Canine Kidney Epithelial Cells grown as 3D cysts in Collagen Type I (7 days old) were exposed to six different concentrations of Hepatocyte Growth Factor (HGF) (0, 1.03, 2.07, 4.15, 8.33 and 16.67 ng/ml). Total RNA was isolated from the cysts after 12 hours of HGF induction. The data submitted here are the raw sequence files of the single read lengths of 50 bp for the 12 samples (2 replicates X 6 conditions) after RNA sequencing experiment using Illumina HiSeq 2000.15
2016-01-17 | E-MTAB-4959 | biostudies-arrayexpress
Project description:Hepatocyte growth factor (HGF) is a well-known promoter of cell proliferation, morphogenesis, migration, angiogenesis and pigmentation. To characterize its effects on human scalp dissected hair follicles, we performed gene expression profiling on RNA isolated from entire hair follicles, comparing the effects of 6hr HGF treatment versus control. 183 genes were differentially up-/down-regulated compared to vehicle control. These genes were involved in a number of different biological pathways suggesting that HGF can have various effects upon human hair follicle function.
2023-05-29 | GSE151560 | GEO
Project description:We developed a post-menopausal orthotopic ER+ breast cancer model that we used to investigate mechanisms of obesity mediated lung metastases. Obesity altered the immune cell composition and function in the primary tumor microenvironment. To identify tumor-derived genes associated with obesity mediated lung metastases we performed RNASeq on tumors grown in lean vs obese mice. From these results we identified a hepatocyte growth factor (HGF)-c-Met (HGF receptor)-CCL2 loop recruiting metastasis-promoting cells.
2022-12-31 | GSE151866 | GEO
Project description:Analysis of Bone Marrow derived macrophages (BMDMs) incubated in presence of Lipopolysaccharide (LPS) (10ng/ml) + Interferon -gamma (IFN-g) (20ng/ml) for 8 h vs Mock treated controls
2017-05-01 | GSE89559 | GEO
Project description:The goal was to perform global transcriptome analysis on MGAS10870 serotype to investigate and compare the gene alterations of WT grown in the presence of 125µg/ml human calprotectin protein to WT grown in the absence of 125µg/ml human calprotectin protein
2020-02-28 | GSE142458 | GEO
Project description:Effect of Hepatocyte Growth Factor (HGF) on lymphatic endothelial cells.
2005-02-01 | E-MEXP-256 | biostudies-arrayexpress