GPR143 regulates ESCRT-dependent exosome biogenesis and promotes cancer metastasis (exosome proteomics)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Q Exactive
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Epithelial Cell, Cell Culture
DISEASE(S): Breast Cancer
SUBMITTER: 
YU JIN LEE
LAB HEAD: Young Chan Chae
PROVIDER: PXD031570 | Pride | 2022-12-20
REPOSITORIES: Pride
Similar Datasets
Project description:Exosomes transport a variety of macromolecules and modulate intercellular communication in physiology and disease. However, the regulation mechanisms that determine exosome contents during exosome biogenesis remain poorly understood. Here, we identify GPR143, an atypical GPCR, as a regulator of the endosomal sorting complex required for transport (ESCRT)-dependent exosome biogenesis pathway. GPR143 interacts with HRS (an ESCRT-0 Subunit) and promotes its association to cargo proteins, such as EGFR, which subsequently enables selective protein sorting into intralumenal vesicles (ILVs) in multivesicular bodies (MVBs). GPR143 is elevated in multiple cancers and quantitative proteomic and RNA profiling of exosomes revealed the GPR143-ESCRT pathway promotes secretion of exosomes that carry unique cargo, including integrins signaling proteins. By gain- and loss-of-function studies, we reveal GPR143 promotes metastasis by secreting exosomes and increasing cell motility/invasion through the integrin/FAK/Src pathway. These finding uncover a mechanism that regulates the exosomal proteome and demonstrate its ability to promote cancer metastasis.
2022-12-20 | PXD031515 | 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
Project description:Contacts between organelles create microdomains that play major roles in regulating key intracellular activities and signaling pathways, but whether they also directly regulate systemic cellular activities, remains unknown. Here, we report the ultrastructural organization and dynamics of a new type of inter-organelle contact which is formed by rough-Endoplasmic Reticulum that is closely wrapped around the mitochondrion (wrappER). To elucidate the in vivo function of this inter-organelle association, mouse liver fractions enriched in wrappER-associated-mitochondria were analyzed in parallel by transcriptomics, proteomics, and lipidomics. The biochemical signature of the wrappER points to an unexpected role in the biogenesis of very-low-density lipoproteins (VLDLs). Altering wrappER-mitochondria contacts curtails liver VLDL secretion and increases hepatic fatty acids, lipid droplets and neutral lipid content. Conversely, acute liver-specific ablation of Mttp, the most upstream regulator of VLDL biogenesis, mirrors this hepatic dyslipidemia phenotype and promotes remodelling of the wrappER-mitochondria contact. The participation of liver wrappER-mitochondria contacts in VLDL biology reveals a role of inter-organelle contacts in systemic lipid homeostasis.
2021-03-15 | PXD021826 | Pride
Project description:The aim of the proteomics experiment was to determine the specificity of the generated monobodies for the target protein STAT3. This was done in an affinity-enrichment experiment using two endogenously expressed and tandem-affinity-tagged monobodies in two different cell lines (Jurkat and K562). Eluted proteins after affinity purification were resolved by SDS-PAGE, and analysed using bottom-up proteomics workflow including tryptic digestion, LC-MS/MS, database-search and spectral counting.
2020-07-10 | PXD018374 | Pride
Project description:The serotonin (5-HT) transporter (SERT) is a key determinant of the concentration of 5-HT available for synaptic signaling and is the target of the antidepressant-selective 5-HT reuptake inhibitors (SSRIs). In addition to the role of SERT in mediating the actions of antidepressant medications, multiple, rare gain of function coding variants in SERT have been identified in subjects with autism spectrum disorder (ASD). The Blakely lab has extensively characterized the structural and functional impact of one of these variants, SERT Ala56, in vitro and in vivo, revealing a constitutively-imposed, biased conformation that enhances transporter function and that perturbs transporter regulation. We hypothesize that SERT Ala56 disrupts the interactions of molecular networks that normally ensure proper transporter structure, localization and regulation, and whose elucidation may reveal new determinants of serotonergic dysfunction in ASD. Here, we pursue an unbiased proteomic analysis of changes in SERT interacting protein (SIPs) imposed by the SERT Ala56 variant, expressed in transgenic mice. Further investigation of the SIPs and associated networks nominated through our studies may provide new opportunities to detect and modulate brain networks that contribute to the underlying complexity of ASD.
2020-05-29 | PXD018695 | Pride
Project description:This project describes the isoniazid (INH) resistance acquisition event in Mycobacterium tuberculosis (Mtb) from the proteomics perspective. In this way, an exploration of the protein differences, comparing clonal INH susceptible (INHs) and INHr pairs of Mtb were evaluated. One clonal clinical and one clonal laboratory-derived Mtb pair with different susceptibility profiles to INH were studied. The laboratory INHr strain had one katG mutation (V1A), while the clinical INHr strain had two (V1A and E3V). Large-scale bacterial cultures were grown in triplicate to obtain secreted proteins as well as proteins from cell fractions. The resulting peptide solutions from all fractions were analyzed using liquid-chromatography coupled with tandem mass spectrometry (LC-MS/MS). LC-MS/MS spectra were compared against an Mtb database to determine the protein abundance. Protein abundance differences were tested by Student’s t test. Looking at the same cellular fractions, there were 25 commonly altered Mtb proteins after acquiring INH resistance. These proteins were involved in ATP synthase machinery, lipid metabolism, regulatory events, virulence, detoxification and adaptation processes.
2018-06-25 | PXD009549 | Pride
Project description:This project is a report of a chromosome-based human proteome project focused on chromosome 9 (Chr 9). To reveal missing proteins and undiscovered features in proteomes, LC-MS/MS analysis based identification and characterization were conducted on 5 pairs of lung adenocarcinoma tumors and adjacent non-tumor tissues.
2015-07-31 | PXD002523 | Pride
Project description:Lipid transporters from the major facilitator superfamily (MFS) have been reported to play central roles in diverse fundamental biological pathways. However, little is known about their transport mechanisms and only the structure of the MFS lipid transporter LtaA in an outward-facing conformation has been determined. LtaA was shown to be a proton-dependent lipid transporter, essential for anchoring of lipoteichoic acids (LTA) to the plasma membrane of the pathogenic bacteria Staphylococcus aureus. Here we used a ‘repeat swap’ approach in combination with cross-linking analysis to predict an inward-facing state of LtaA, performed molecular dynamics simulations, in vitro and in vivo activity assays, and mutagenesis analysis to characterize outward- and inward-facing conformations of LtaA in lipid membranes. Our results indicate that LtaA catalyze lipid translocation by a “trap-and-flip” mechanism, paving the way for the mechanistic characterization of other MFS lipid transporters.
2022-05-16 | PXD031166 | Pride
Project description:Endometriosis is a debilitating gynecological disorder affecting approximately 10% of the female population. Despite its prevalence, robust methods to classify and treat endometriosis remain elusive. Changes throughout the menstrual cycle in tissue size, architecture, cellular composition, and individual cell phenotypes make it extraordinarily challenging to identify markers or cell types associated with uterine pathologies since disease-state alterations in gene and protein expression are convoluted with cycle phase variations. Here, we developed an integrated workflow to generate both proteomic and single-cell RNA-sequencing (scRNA-seq) data sets using tissues and cells isolated from the uteri of control and endometriotic donors. Using a linear mixed effect model (LMM), we identified proteins associated with cycle stage and disease, revealing a set of genes that drive separation across these two biological variables. Further, we analyzed our scRNA-seq data to identify cell types expressing cycle and disease- associated genes identified in our proteomic data. A module scoring approach was used to identify cell types driving the enrichment of certain biological pathways, revealing several pathways of interest across different cell subpopulations. Finally, we identified ligand-receptor pairs including Axl/Tyro3 – Gas6, that may modulate interactions between endometrial macrophages and/or endometrial stromal/epithelial cells. Analysis of these signaling pathways in an independent cohort of endometrial biopsies revealed a significant decrease in Tyro3 expression in patients with endometriosis compared to controls, both transcriptionally and through histological staining. This measured decrease in Tryo3 in patients with disease could serve as a novel diagnostic biomarker or treatment avenue for patients with endometriosis. Taken together, this integrated approach provides a framework for integrating LMMs, proteomic and RNA-seq data to deconvolve the complexities of complex uterine diseases and identify novel genes and pathways underlying endometriosis.
2023-09-05 | PXD045115 | Pride
Project description:Mitochondrial oxidation-induced cell death, a physiological process triggered by various cancer therapeutics to induce oxidative stress on tumours, has been challenging to investigate owing to the difficulties in generating mitochondria-specific oxidative stress and monitoring mitochondrial responses simultaneously. Accordingly, to the best of our knowledge, the relationship between mitochondrial protein oxidation via oxidative stress and the subsequent cell death-related biological phenomena has not been defined. Here, we developed a multifunctional iridium(III) photosensitiser, Ir-OA, capable of inducing substantial mitochondrial oxidative stress and monitoring the corresponding change in viscosity, polarity, and morphology. Photoactivation of Ir-OA triggers chemical modifications in mitochondrial protein-crosslinking and oxidation (i.e., Oxidative phosphorylation complexes and channel and translocase proteins), leading to microenvironment changes, such as increased microviscosity and depolarisation. These changes are strongly related to cell death by inducing mitochondrial swelling with excessive fission and fusion. We suggest a potential mechanism from mitochondrial oxidative stress to cell death based on proteomic analyses and phenomenological observations.
2021-03-23 | PXD022163 | Pride