Reference Proteome-Based Characterization of Liver Cancer
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo Sapiens (human)
SUBMITTER: Jinhwan Eugene Lee
PROVIDER: PXD027081 | JPOST Repository | Mon Jul 04 00:00:00 BST 2022
REPOSITORIES: jPOST
Similar Datasets
Project description:Metabolic rewiring is essential for breast tumor growth and progression to metastatic disease, yet little is known regarding how cancer cells modify their acquired metabolic programs in response to different metastatic microenvironments. Transcriptional and metabolomic analysis have previously shown that liver-metastatic breast cancer cells adopt an intrinsic metabolic profile characterized by increased HIF-1α activity and dependence on glycolysis. Stable isotope tracing analysis (SITA) performed in vivo confirmed that the glycolytic nature of liver-metastatic breast cancer cells is retained when these cells are grown as primary tumors or as liver metastases. However, our data also reveal that unique metabolic adaptations are specifically induced by the liver microenvironment. Indeed, liver metastases display elevated expression of genes involved in glutathione (GSH) biosynthesis and reactive oxygen species (ROS) detoxification when compared to primary tumors. Moreover, breast cancer liver metastases rely strongly on glucose and glutamine-derived carbons to support de novo GSH synthesis. Glutathione is a tripeptide that acts as a major scavenger for reactive oxygen species (ROS). Liver metastases display reduced levels of 8-Oxo-2'-deoxyguanosine, confirming their increased capacity to buffer ROS. We demonstrated the dependence of liver metastases formation on these distinct metabolic adaptations by depleting glutamate-cysteine ligase (Gcl), the rate-limiting enzyme in glutathione biosynthesis. Gcl depletion, and decreased GSH levels, strongly reduced the capacity of liver-metastatic cells to form distant metastasis within the liver. We performed gene expression analysis of liver metastasis and primary tumors from liver-metastatic breast cancer cells derived from 4T1 cells. 4T1-2776 (76) and 4T1-2792 (92) cells were injected in the mammary fat pads of BALB/c and grown as primary tumors or were grown as liver metastasis after splenic injection. Tissues were harvested from primary tumors and liver metastasis and submitted to laser-capture microdissection (LCM). For liver metastasis, tumour tissue was harvested at 10 days, 2 weeks and 3 weeks after splenic injection. For each time point, tissue was isolated from the tumor centre (core), a peripheral area of the tumor (margin), a region of the liver proximal to the tumor (adjacent) and a region of the liver far from the tumor (distant). For primary tumors from 4T1-2776 cells (1034, 1036, 1043) and 4T1-2792 (MFP2L, MFP3L, MFP5L) tissue was collected from the tumor core and margin, at the experimental endpoint.
2023-07-25 | GSE238214 | GEO
Project description:To dissect how diurnal rhythms affect key functions such as transcription or chromatin remodelling, we quantified the temporal nuclear accumulation of proteins and phosphoproteins from mouse liver using in vivo stable isotope labelling by amino acids in cell culture (SILAC)-based mass spectrometry (MS). Protein extracts from isotope labelled mice liver nuclei were used as a reference and mixed with extracts from animals collected every 3 h for 45 h total. Phosphopeptide levels were analysed after enrichment with titanium dioxide (TiO2). The proteins levels were also analysed (dataset with project accession PXD003818).
2016-12-22 | PXD004191 | Pride
Project description:Colorectal cancer (CRC) is the third most common and second most deadly cancer worldwide. Treatments for metastatic CRC (mCRC) are still largely based on toxic chemotherapy regimens, but important insights into tumor biology enabled the development of targeted cancer therapies. Here, we describe a proteogenomic analysis of CRC liver metastases (metastatic CRC, mCRC), an ideal setting to analyze therapeutic resistance which occurs in a short time frame. We selected liver metastases from two CRC patients on both of which we performed deep proteome profiling and WES and RNAseq-directed database searches, identifying 10 predicted muttaions, one of which was KRAS G12V. finding we generated targeted parallel reaction monitoring (PRM) assays using stable isotope labelled standard (SIS) peptides to quantify the actual concentration (fmol per µg of total protein) of the mutated and canonical (wildtype) protein variants for 8 different mutations. We demonstrate on a total of 8 tumor and 6 paired healthy tissue samples (7 and 6 out of these KRASG12V) that PRM allows quantifying the actual mutation rate on the protein level from as little 10 µg of total protein starting amount and within a single 1 hour nano-LC-MS/MS run.
2019-12-04 | PXD014222 | Pride
Project description:The RNA-seq technique was used to sequence the liver cancer tissues and paracancerous tissues of 6 clinical patients, aiming to find differentially expressed genes and transcripts, and to verify the expression level and function of the expression. To improve the accuracy of gene discovery, we used two methods to analyze the sequencing data.
2019-09-05 | GSE136846 | GEO
Project description:Stable isotope profiling of ETBE degraders
| PRJEB52346 | ENA
Project description:Stable isotope probing of kangaroo forestomach content
| PRJNA230753 | ENA
Project description:Acetylation of lysine is involved in various biological processes and is considered as a key reversible post-translational modification (PTM) in the regulation of gene expression, enzyme activity, protein-protein interaction, and subcellular localization. This PTM is therefore highly relevant in the context of circadian biology but its characterization at the genome wide scale and its clock-dependent or -independent regulation is still poorly characterized. Here we used in vivo stable isotope labeling by amino acids (SILAC) coupled with an enrichment of acetylated-lysine-containing peptides to provide a comprehensive and rhythmic acetylome map of the liver nucleus.
2017-08-02 | PXD005310 | Pride
Project description:Acetylation of lysine is involved in various biological processes and is considered as a key reversible post-translational modification (PTM) in the regulation of gene expression, enzyme activity, protein-protein interaction, and subcellular localization. This PTM is therefore highly relevant in the context of circadian biology but its characterization at the genome wide scale and its clock-dependent or -independent regulation is still poorly characterized. Here we used in vivo stable isotope labeling by amino acids (SILAC) coupled with an enrichment of acetylated-lysine-containing peptides to provide a comprehensive and rhythmic acetylome map of the liver.
2017-08-02 | PXD005317 | Pride
Project description:Phosphohistidine signaling in mammalian biological systems has not been well characterized. In this study, we investigated the impact of phosphohistidine phosphatase 1 (PHPT1) expression on the hepatic proteome profile in vivo. PHPT1 overexpression was carried out using adenoviral-based transgene delivery for liver-specific overexpression of PHPT1 while a heterozygous knockout was used as a model of PHPT1 deficiency. These two models were characterized using mass spectrometry-based proteomics and label-free relative quantitation. In conjunction with bioinformatic analysis, PHPT1-induced changes in several key proteins and pathways were identified in the liver. Implementation of these models in future studies will lead to far greater insight into the role of phosphohistidine signaling and PHPT1 regulation in mammalian cells.
2020-07-29 | PXD008323 | Pride