Metabolomic and Proteomic Analysis of Breast Cancer Exosomes Reveals Alterations in Purine and Carnitine Metabolism
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Orbitrap Fusion
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Epithelial Cell, Breast Cell Line
DISEASE(S): Breast Cancer
SUBMITTER: 
Abdullah Khraibah
LAB HEAD: Maxim Berezovski
PROVIDER: PXD056161 | Pride | 2025-03-25
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| MCF10_1_A.raw | Raw | |||
| MCF10_1_B.raw | Raw | |||
| MCF10_2_A.raw | Raw | |||
| MCF10_2_B.raw | Raw | |||
| MCF10_3_A.raw | Raw |
Items per page: 5 1 - 5 of 15 |
Similar Datasets
Project description:The biological and psychological importance of hair is recognized worldwide. Molecules that can promote the activation of hair follicle stem cells and initiation of the growth phase have been the subject of much research. Clarifying how hair regeneration is regulated may help to provide hair loss treatments, including cosmetic and even psychological interventions. This study focuses on the hair-growing effects of a cell extract (CE) obtained from the cactus Notocactus ottonis by a cold vacuum extraction protocol, with the aim of investigating its hair-growing effects and relevant mechanisms and potential factors therein. In an experiment using female C57BL/6 mice, a vehicle control group (VC: propylene glycol : ethanol : water), a positive control group (MXD : 3% minoxidil), and an experimental group (N-CE: 10% Notocactus ottonis CE) were applied topically to the backs of mice. Results showed that 3% MXD and 10% N-CE were more effective in promoting hair growth than the VC. Furthermore, an increase in the number of hair follicles was observed with 10% N-CE CE in hematoxylin-eosin-stained skin tissue. We also examined the metabolite composition of the N-CE. Additionally, the whole genome DNA microarray and total protein profiling was carried out on the skin samples to suggest molecular factors underlying the hair-promoting effects of N-CE. This study raises the possibility of using the newly described Notocactus ottonis cell extract as a hair-growth promoting agent.
2023-03-11 | PXD039218 | Pride
Project description:Biomarker identification for diagnosis of systemic juvenile idiopathic arthritis (SJIA), an auto-inflammatory disease that presents with prolonged fevers. Disease vs. healthy control
2015-09-03 | E-GEOD-57183 | biostudies-arrayexpress
Project description:In this work, we developed an ultra-sensitive CE-MS/MS method for bottom-up proteomics analysis of limited samples, down to sub-nanogram levels of total protein. Analysis of 880 pg and 88 pg of HeLa protein digest standard by CE-MS/MS yielded ~1,100±46 and ~160±59 proteins, respectively, demonstrating higher protein and peptide identifications than the current state-of-the-art CE-MS/MS-based proteomic analyses with similar amounts of sample. To demonstrate potential applications of our ultra-sensitive CE-MS/MS method for analysis of limited biological samples, we digested 500 and 1,000 HeLa cells using a miniaturized in-solution digestion workflow. From 1-, 5-, and 10-cell equivalents injected from the resulted digests, we identified 744±127, 1,139±24, and 1,271±6 proteins and 3,353±719, 5,709±513, and 8,527±114 peptide groups, respectively. Furthermore, we performed a comparative assessment of CE-MS/MS and two reversed-phased nano-liquid chromatography (RP-nLC-MS/MS) methods (monolithic and packed columns) for the analysis of a ~10 ng HeLa protein digest standard. Our results demonstrate complementarity in the protein-, and especially peptide-level identifications of the evaluated CE-MS- and RP-nLC-MS-based methods. The techniques were further assessed to detect post-translational modifications and highlight the strengths of the CE-MS/MS approach in identifying potentially important and biologically relevant modified peptides. With a migration window of ~60 min, CE-MS/MS identified ~2,000±53 proteins on average from a single injection of ~8.8 ng HeLa protein digest standard. Additionally, an average of 232±10 phosphopeptides and 377±14 N-terminal acetylated peptides were identified in CE-MS/MS analyses at this sample amount corresponding to 2- and 1.5-fold more identifications for each respective modification found by nLC-MS/MS methods.
2022-02-17 | PXD027226 | Pride
Project description:The importance of the mammalian intestinal microbiota to human health has been intensely studied over the past few years. It is now clear that the interactions between human hosts and their associated microbial communities need to be characterized in molecular detail if we are to truly understand human physiology. Additionally, the study of such host-microbe interactions is likely to provide us with new strategies to manipulate such complex systems to maintain or restore homeostasis in order to prevent or cure pathological states. We describe the use of high-throughput metabolomics to shed light on the interactions between the intestinal microbiota and the host. We show that treatment with the antibiotic streptomycin disrupts intestinal homeostasis and has a profound impact on the intestinal metabolome, affecting the levels of over 87% of all metabolites detected. Many metabolic pathways that are critical for host physiology were affected, including bile acid, eicosanoid and steroid hormone synthesis. Interestingly, many of these pathways are also affected by intestinal pathogens. Dissecting the effect of both beneficial and pathogenic bacteria on some of these pathways will be instrumental in understanding the interplay between the host, the resident microbiota and incoming pathogens and may aid in the design of new therapeutic strategies that target these interactions. Age-matched female C57BL/6 mice were used. Fresh feces were collected and stored at -80 oC. Mice were then treated with 20 mg of streptomycin through oral gavage and fresh feces were collected 24 hours after treatment and stored at -80 oC until used. To extract metabolites from feces, acetonitrile was added to samples (approximately 10-25 μL of acetonitrile per 1 mg of tissue), which were then homogenized. The samples were then cleared by centrifugation and the supernatant was collected and dried at room temperature using a centrifuge equipped with a vacuum pump. All extracts were kept at -80 oC until used. For metabolic profiling, the dried extracts from mouse feces were suspended in a 2:3 mixture of water and acetonitrile (10 μL per 1 mg of tissue), vortexed and cleared by centrifugation. Supernatants were collected and used as described below. Extracts were diluted 1:5 with 60% acetonitrile containing either 0.2% formic acid (for positive ion mode) or 0.5% ammonium hydroxide (for negative ion mode) and spiked with predefined amounts of the "ES tuning mix" solution as the internal standard for mass calibration. The solutions were then infused, using a syringe pump (KDS Scientific, Holliston, USA) at a flow rate of 2.5 µL per minute, into a 12-T Apex-Qe hybrid quadrupole-FT-ICR mass spectrometer (Bruker Daltonics, Billerica, USA) equipped with an Apollo II electrospray ionization source, a quadrupole mass filter and a hexapole collision cell. Data were recorded in positive and negative ion modes with broadband detection and an FT acquisition size of 1024 kilobytes per second, within a range of m/z 150 to 1100. Under these settings, a mass resolution of ca. 100,000 (full width at half maximum, FWHM) at m/z 400 and a mass accuracy within 2 ppm or less for all detected components, following internal mass calibration, were observed. Other experimental parameters were: capillary electrospray voltage of 3600-3750 V, spray shield voltage of 3300-3450 V, source ion accumulation time of 0.1 s and collision cell ion accumulation time of 0.2 s. To increase detection sensitivity, survey scan mass spectra in positive and negative ion modes were acquired from the accumulation of 200 scans per spectrum, and duplicate acquisitions per sample were carried out to ensure data reproducibility. Raw mass spectrometry data were processed as described elsewhere (Han et al. 2008. Metabolomics. 4:128-140). To identify differences in metabolite composition between untreated and treated samples, we first filtered our list of masses for metabolites that were present on one set of samples (untreated or treated) but not the other. Additionally, we averaged the mass intensities of metabolites in each group and calculated the ratios between averaged intensities of metabolites from untreated and treated samples. To assign possible metabolite identities to the masses present in only one of the sample groups or showing at least a 2-fold change in intensities between the sample groups, the monoisotopic neutral masses of interest were queried against MassTrix (http://masstrix.org), a free-access software designed to incorporate masses into metabolic pathways. Masses were searched against the Mus musculus database within a mass error of 3 ppm. Data were analyzed by unpaired t tests with 95% confidence intervals.
2011-02-08 | E-GEOD-25687 | biostudies-arrayexpress
Project description:A human microarray comprising 1658 human genome probes was used to evaluate the specific expression of miRNA between brian glioma stem cells (GSC) and normal neural stem cells(NSC). 2 total RNA samples are analyzed, brian glioma stem cells (GSC) and normal neural stem cells(NSC). cell type: brain glioma stem cells:GSC_1, GSC_2, GSC_3 ; cell line: normal neural stem cells: NSC_1, NSC_2, NSC_3 biological replicate: NSC_1, NSC_2, NSC_3; biological replicate: GSC_1, GSC_2, GSC_3
2014-07-30 | E-GEOD-41032 | biostudies-arrayexpress
Project description:Primary tumor recurrence occurs commonly after surgical resection of lung squamous cell carcinoma (SCC). The aim of this study was to identify genes involved in recurrence in lung squamous cell carcinoma patients. Array comparative genomic hybridization (aCGH) was performed on DNA extracted from tumour tissue from 62 patients with primary lung squamous cell carcinomas. aCGH data was analysed to identify genes affected by copy number alterations that may be involved in SCC recurrence. Candidate genes were then selected for technical validation based on differential copy number between recurrence and non-recurrence SCC tumour samples. Genes technically validated advanced to tests of biological replication by qPCR using an independent test set of 72 primary lung SCC tumour samples. 18q22.3 loss was identified by aCGH as significantly associated with recurrence (p=0.038). Although aCGH copy number loss associated with recurrence was found for seven genes within 18q22.3, only SOCS6 copy number loss was both technically replicated by qPCR and biologically validated in the test set. DNA copy number profiling using 44K element array comparative genomic hybridization microarrays of 62 primary lung squamous cell carcinomas.
2011-12-25 | E-GEOD-32058 | biostudies-arrayexpress
Project description:Coelomic epithelium (CE), or coelothelium, is a mesodermal-derived tissue lining coelomic cavities and enveloping inner organs in all coelomates, including vertebrates, where it forms both peritoneum and pericardium, as well as pleura, in mammals. Due to its histogenetic potential, CE is considered a key-tissue characterized by unique features. At present, in adult echinoderms, CE is supposed to play fundamental key roles acting as: 1) source of stem/multipotent cells (undifferentiated coelomocytes), particularly during regenerative phenomena (Candia Carnevali et al., 2009; Candia Carnevali and Burighel, 2010; Hernroth et al., 2010; García-Arrarás et al., 2011). 2) haematopoietic tissue, being the direct source of circulating elements (differentiated coelomocytes) including immune cells, indispensable elements during the first repair events of clotting and defence against pathogens (Boassche and Jangoux, 1976; Munoz-Chapuli et al., 2005; Pinsino et al., 2007). Despite these indications, the histological approach alone, microscopic or ultrastructural, is not sufficient to fully attribute a haematopoietic role to the CE and even the presumptive production/release process of stem/pluripotent cells from coelothelium during regeneration need to be confirmed as well. As a matter of fact, it is still unclear if the intense proliferation detectable in regenerating CE is functional to the production of wandering coelomocytes necessary to restore the post-traumatic fluid loss or rather to produce truly regeneration-competent cells used as building blocks of the regenerating tissues. In order to overcome some of these issues and deeply understand the complex physiological role of this epithelial tissue in echinoderm biology, it is important to integrate morphological evidences with detailed molecular data. In particular, proteomics is becoming a powerful tool to deepen the problem of cell and tissue functions during both tissue homeostasis and regeneration by depicting the existing protein activities involved in the regrowth processes. However, proteomics investigations focussed on echinoderm CE are quite limited. Specific molecular information are due to the works by Gabre and co-workers (2015) and by Kim and co-workers (2018), which are addressed to the characterization of CE transcriptome from the clonal starfish Coscinasterias muricata and the blue bat starfish Patiria pectinifera, respectively. The analysis of the biological processes associated to the transcripts identified in these studies suggests the involvement of CE in the development of anatomical units. More recently the CE proteome of the common starfish Asterias rubens was also published (Sharlaimova et al., 2021). Despite these evidences, there is a great lack of integrated multidisciplinary studies that can contribute to unravel the complex roles exerted by this tissue both in regeneration and other physiological processes. In order to partially unravel this gap, the present work is addressed to analyse the CE of the starfish Marthasterias glacialis in both standard physiological (homeostatic) and regenerating conditions, by combining histological/ultrastructural analyses with appropriate proteomics approaches, the former to provide a detailed cell and tissue perspective, the latter to contribute a base-line molecular view of CE multifunctional implications. This integrated approach can significantly help to shed new light into the intriguing role of this apparently simple but fundamental tissue.
2023-01-24 | PXD027009 | Pride
Project description:In studies presented here, recombinant human serum albumins (rHSA) produced in Oryza sativa (Asian rice) (OsrHSA) from a number of suppliers have been extensively characterized and compared to plasma-derived HSA (pHSA) and rHSA expressed in yeast (Pichia pastoris and Saccharomyces cerevisiae). Modifications of the samples were identified by liquid chromatography-mass spectrometry (LC-MS) with samples further assessed with size exclusion chromatography (SEC), reversed-phase high-performance liquid chromatography (RP-HPLC), capillary electrophoresis (CE) as well as additionally biophysical techniques. LC-MS analysis identified a greater number of hexose-glycated arginine and lysine residues on OsrHSA compared to pHSA or rHSA expressed in yeast. Lot-to-lot variability of OsrHSA glycation from one manufactures supplied products were further identified. The extensive glycation of OsrHSA from multiple suppliers may have further implications for the use of OsrHSA as a therapeutic product.
2014-09-12 | PXD001248 | Pride
Project description:BackgroundHuR/ELAV1, a ubiquitous RNA-binding protein, belongs to the RNA-binding protein family and is crucial for stabilizing and regulating the translation of various mRNA targets, influencing gene expression. Elevated HuR levels are associated with multiple disorders, including cancer and neurodegenerative diseases. Despite the identification of small molecule inhibitors targeting HuR, their detailed characterization remains limited. Recently, Eltrombopag, an FDA-approved drug for immune thrombocytopenic purpura and chemotherapy-induced thrombocytopenia, emerged as a potential HuR inhibitor. However, the specific molecular pathways influenced by both HuR and Eltrombopag are not fully understood. ResultsOur study demonstrates that Eltrombopag operates via HuR inhibition, affecting gene expression regulation at the posttranscriptional level. We show that both HuR knockout and Eltrombopag treatment modulate iron metabolism by decreasing ferritin heavy chain (FTH1) and light chain (FTL) synthesis while increasing the expression of Iron-regulatory protein 2 (IRP2), a key regulator of ferritin translation. Additionally, HuR inhibition reduces the levels of Glycoprotein Hormones, Alpha Polypeptide (CGA), a marker associated with hormone-induced tumors, suggesting a potential use of Eltrombopag in treatment of cancers overexpressing CGA. We observed that the main of control is manifested at the level of translation inhibition, with proteosome-mediated regulation also playing an important role.ConclusionsThese findings uncover novel posttranscriptional mechanisms governed by HuR and its inhibitor, elucidating pathways relevant to HuR-mediated regulation and molecular therapies aimed at targeting this protein.
2024-12-20 | PXD050698 | Pride
Project description:Functional oncogenic links between ErbB2 and ERR⍺ in HER2+ breast cancer patients support a therapeutic benefit of co-targeted therapies. However, ErbB2 and ERR⍺ also play key roles in heart physiology, and this approach could pose a potential liability to cardiovascular health. Using integrated phosphoproteomic, transcriptomic and metabolic profiling, we uncovered molecular mechanisms associated with the adverse remodeling of cardiac functions in mice with combined attenuation of ErbB2 and ERR⍺ activity. Genetic disruption of both effectors results in profound effects on cardiomyocyte architecture, inflammatory response and metabolism, the latter leading to a decrease in fatty acyl-carnitine species further increasing the reliance on glucose as a metabolic fuel, a hallmark of failing hearts. Furthermore, integrated omics signatures of ERR⍺ loss-of-function and doxorubicin treatment exhibit reciprocal features of chemotherapeutic cardiotoxicity. These findings thus reveal potential cardiovascular risks in discrete combination therapies in the treatment of breast and other cancers.
2022-09-22 | PXD032766 | Pride