Inhibiting TGFβ-1 pathway reduces the aggressiveness of intrahepatic CCA HuCCT1 CD90 positive cells
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1248463 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| Other | ||||
| SRR33050987.fastq.gz | Fastqsanger.gz | |||
| SRR33050988.fastq.gz | Fastqsanger.gz | |||
| SRR33050989.fastq.gz | Fastqsanger.gz | |||
| SRR33050991.fastq.gz | Fastqsanger.gz |
Items per page: 5 1 - 5 of 6 |
Similar Datasets
Project description:Molecular mechanisms responsible for the poor prognosis in patients with intrahepatic cholangiocarcinoma (CCA) are still unknown, however, the stem cell marker cluster differentiation 90 (CD90), has been reported to be associated with a more aggressive cancer phenotype. In this scenario, TGFβ1 signaling pathway likely has a role as master gene regulator. Aim of the study is to investigate the role of CD90 in iCCA aggressiveness. The molecular profile of HuCCT1/CD90+ and HuCCT1/CD90- cells was obtained through transcriptomic analysis (NGS). Bioinformatic data were confirmed in both cell lines by qRT-PCR and western blot. Cells were treated with Gemcitabine in monotherapy or in combination with Galunisertib, a selective TGFβRI inhibitor in 2D and 3D models. HuCCT1 CD90 positive cells are more proliferative, less migratory, and resistant to Gemcitabine treatment. Next, HuCCT1/CD90+ express lower levels of TGFβ1 compared to /CD90- cell lines. Finally, HuCCT1/CD90+ are resistant to Gemcitabine, while the combination of both Gemcitabine and Galunisertib displays a synergistic effect on HuCCT1/CD90+ cell proliferation. These results underline that CD90 inducing Gemcitabine resistance can be overcome by adding a TGFβ1 inhibitor such as Galunisertib, thereby moving further toward a precision medicine approach in patients with iCCA.
2025-06-18 | GSE294148 | GEO
Project description: Not available
| S-EPMC12155448 | biostudies-literature
Project description:Intrahepatic Cholangiocarcinoma (iCCA) is a difficult type of cancer to diagnose, extremely aggressive and resistant to therapeutic options with an increased incidence and mortality over time. The overexpression of Notch pathway is implicated in iCCA to create an ecosystem that drives the tumor aggressiveness. Specifically, Notch1 is overexpressed in iCCA tissue of patients, playing an important role on tumor growth. Our purpose was to investigate the effectiveness of Crenigacestat in in vivo experiments, using Hucct1 injected in CD1 mice to generate an iCCA xenograft mouse model. In the present study, in order to explore modulated factors and genes by treatment we performed a transcriptomic analysis validated by Western Blotting and qRT-PCR on iCCA tumor masses of xenograft mice. Our results indicate that the treatment significantly inhibited Notch1 and HES1 genes and proteins triggering a strong immune response. Nevertheless, moderate fibrosis was shown in this model which may have hindered response to therapy to promote tumor progression. We used microarray technology to understand the molecular mechanisms affected by Crenigacestat in HUCCT1 xenograft model of intraepatic cholangiocarcinoma.
2022-04-27 | GSE150024 | GEO
Project description:The biological significance of the down-regulation of miR-376c in HuCCT1 cells is unknown. We revealed that miR-376c significantly reduced EGF-dependent cell migration through its target GRB2 in HuCCT1 cells. To elucidate key molecules in EGF-dependent HuCCT1 migration, in which GRB2 upregulation is caused by abnormal suppression of miR-376c, we compared expression profiles of mRNAs affected by pre-miR-376c and siRNA targeting GRB2.
2013-05-23 | GSE47186 | GEO
Project description:The biological significance of the down-regulation of miR-376c in HuCCT1 cells is unknown. We revealed that miR-376c significantly reduced EGF-dependent cell migration through its target GRB2 in HuCCT1 cells. To elucidate key molecules in EGF-dependent HuCCT1 migration, in which GRB2 upregulation is caused by abnormal suppression of miR-376c, we compared expression profiles of mRNAs affected by pre-miR-376c and siRNA targeting GRB2. We compared expression profiles of mRNAs affected by pre-miR-376c and siRNA targeting GRB2 in EGF-treated HuCCT1 cells.
2013-05-23 | E-GEOD-47186 | biostudies-arrayexpress
Project description:We described differential expression of CD90 in gp38+ fibroblasts in murine colon. We functionally characterized CD90- and CD90+ fibroblasts by co-culturing with intestinal organoids. CD90+ colon fibroblasts were able to support organoid growth while CD90- did not.
2018-10-27 | GSE105102 | GEO
Project description:Cholangiocarcinoma (CCA) is a devastating liver cancer characterized by high aggressiveness and resistance to therapy, which results to poor prognosis. Signals imposed by oncogenic pathways, such as transforming growth factor β (TGFβ) frequently contribute to CCA development. In this study, we explored novel effectors of the TGFβ pathway in CCA by gene expression profiling. We identified the long non-coding RNA LINC00313 as a novel target gene of TGFβ signalling in CCA cells. TGFβ induced LINC00313 expression in a TβRI/Smad-dependent manner. Subcellular fractionation showed that LINC00313 is a predominantly nuclear lncRNA. By integrating RNA-seq and ATAC-seq data from LINC00313 over-expressing cells, we observed that LINC00313 regulates the expression of several genes involved in the Wnt signalling pathway. As a proof of concept, we focused on the gene encoding transcription factor 7 (TCF7), a major effector that drives transcription of Wnt-target genes. LINC00313 gain of function resulted in increased TCF7 expression, while its loss of function diminished basal or TGFβ-induced TCF7 expression levels. Interestingly, LINC00313 enhanced basal or chemically induced TCF/LEF-dependent transcriptional responses, promoted colony-forming capacities of CCA cells in vitro and accelerated tumor growth in vivo. We also report that genes associated with LINC00313 over-expression recapitulate poor prognosis human CCA associated with a reduced overall survival and KRAS mutations. To decipher the underlying molecular functions of LINC00313, we identified its interacting proteins by performing an unbiased RNA pull-down assay followed by mass spectrometry. We demonstrated that actin-like 6A (ACTL6A), a subunit of the SWI/SNF chromatin remodelling complex specifically binds to LINC00313 and impacts TCF7 expression and TCF/LEF signalling output. Thus, we propose a model whereby TGFβ induces LINC00313, in order to regulate the expression of a subset of target genes, such as TCF7 possibly in co-operation with the SWI/SNF chromatin remodelling complex, via establishing direct interaction with ACTL6A. By regulating key genes of the Wnt pathway, LINC00313 fine-tunes Wnt/TCF/LEF-dependent transcriptional responses and boosts cholangiocarcinogenesis.
2024-02-05 | E-MTAB-12879 | biostudies-arrayexpress
Project description:Cholangiocarcinoma (CCA) is a devastating liver cancer characterized by high aggressiveness and resistance to therapy, which results to poor prognosis. Signals imposed by oncogenic pathways, such as transforming growth factor β (TGFβ) frequently contribute to CCA development. In this study, we explored novel effectors of the TGFβ pathway in CCA by gene expression profiling. We identified the long non-coding RNA LINC00313 as a novel target gene of TGFβ signalling in CCA cells. TGFβ induced LINC00313 expression in a TβRI/Smad-dependent manner. Subcellular fractionation showed that LINC00313 is a predominantly nuclear lncRNA. By integrating RNA-seq and ATAC-seq data from LINC00313 over-expressing cells, we observed that LINC00313 regulates the expression of several genes involved in the Wnt signalling pathway. As a proof of concept, we focused on the gene encoding transcription factor 7 (TCF7), a major effector that drives transcription of Wnt-target genes. LINC00313 gain of function resulted in increased TCF7 expression, while its loss of function diminished basal or TGFβ-induced TCF7 expression levels. Interestingly, LINC00313 enhanced basal or chemically induced TCF/LEF-dependent transcriptional responses, promoted colony-forming capacities of CCA cells in vitro and accelerated tumor growth in vivo. We also report that genes associated with LINC00313 over-expression recapitulate poor prognosis human CCA associated with a reduced overall survival and KRAS mutations. To decipher the underlying molecular functions of LINC00313, we identified its interacting proteins by performing an unbiased RNA pull-down assay followed by mass spectrometry. We demonstrated that actin-like 6A (ACTL6A), a subunit of the SWI/SNF chromatin remodelling complex specifically binds to LINC00313 and impacts TCF7 expression and TCF/LEF signalling output. Thus, we propose a model whereby TGFβ induces LINC00313, in order to regulate the expression of a subset of target genes, such as TCF7 possibly in co-operation with the SWI/SNF chromatin remodelling complex, via establishing direct interaction with ACTL6A. By regulating key genes of the Wnt pathway, LINC00313 fine-tunes Wnt/TCF/LEF-dependent transcriptional responses and boosts cholangiocarcinogenesis.
2024-02-05 | E-MTAB-12030 | biostudies-arrayexpress
Project description:Glycosphingolipids (GSL), a class of plasma membrane lipids, have been proposed as markers of cancer stem cells (CSC). In particular, gangliosides (GS), sialic acid-containing GSL, have been investigated for their role in the malignant phenotype of several cancers and in tumor-stem like cells, but there are no data about human cholangiocarcinoma (CCA). Our study aims to provide a GSL and GS profiling of both stem-like subsets and their parental cells in human CCA. Experimentals. Intrahepatic CCA cells (HUCCT1, CCLP1) were used. Stem-like subset was enriched by sphere culture (SPH) and compared to parental cells grown as monolayer (MON). CCA GS patterns were determined by chromatographic analytical procedures, and their molecular species identification was evaluated by feeding cells with 3H-sphingosine. GS role in modulation of stem features was investigated using D- threo-1phenyl-2-palmitoylamino-3-N-morpholine-1-propanol (PPMP, glucosylceramide synthase inhibitor), and CCA GD3S-transfected cells. FACS-sorted GD2+ SPH cells were examined for stem-like gene expression compared to GD2- SPH. GS biosynthesis enzymes were analyzed by RT-qPCR at different times of spherogenesis. Results: In both CCA lines, compared to MON, SPH showed drastic changes in specific sphingolipids (Cer, Gb3, SM), and in the amount of total GS. In contrast to MON, CCA-SPH shows increase content of GM3, reduction of GM2; among complex GS, strongly increase of GD1a and appearance of GD2, a finding also corroborated by high levels of GM3 synthase as well as GD3- and GM2/GD2 synthases expression in CCA-SPH. Notably, sphere-forming ability and expression of CSC-related genes were affected by PPMP. Importantly it has emerged that the cancer stem features related on GD2 availability are not due to the ganglioside GD2 synthase enzyme, but depend on the enzyme GD3S, the synthase that provides the precursor (GD3) of ganglioside GD2. Thus we have stably transfected both CCLP1 and HUCCT1 cells with the GD3S gene. GD3S-transfected MON (MON GD3S+) cells showed enhanced sphere-forming ability in vitro, superior invasive properties, as well as higher drug resistance when treated with cisplatin and oxaliplatin compare to the transfected control. To better clarify the molecular features and the altered pathways in MON GD3S+ CCA cells, a global transcriptomic analysis was performed. Likewise, GD2+ SPH cells were enriched with CSC-markers at protein and gene levels in addition to several genes involved in pluripotency, self-renewal, and EMT, compare to GD2- SPH. Notably, expression of GM2/GD2 synthases was significantly expressed in tumor samples compared to paired non-tumoral liver tissue of CCA patients (n=104) and greatly correlated with presence of satellite nodules, lymph node invasion, and recurrence. Conclusions: We show for the first time that the CCA stem-like properties may be associated with GSL synthetic pathway and pattern. GSL and GS synthases could represent potential markers for CCA.
2025-01-08 | GSE238179 | GEO
Project description:To explore the functional difference between CD90+CD39+ and CD90+CD39- fibroblasts in human hypertrophic scar and normal skin, the gene expresson microarray was performed on Live CD49f- E-Cadherin- Lin- CD45- CD31- CD90+ CD39+ and Live CD49f- E-Cadherin- Lin- CD45- CD31- CD90+ CD39- cells sorted from suspension disgested from three human hypertrophic scar samples; and Live CD49f- E-Cadherin- Lin- CD45- CD31- CD90+ CD39+ cells sorted from suspension disgested from three human normal skin samples
2021-07-05 | GSE158766 | GEO