-
1دورية أكاديمية
المؤلفون: Spiros Georgakis, Katerina Gkirtzimanaki, Garyfalia Papadaki, Hariklia Gakiopoulou, Elias Drakos, Maija-Leena Eloranta, Manousos Makridakis, Georgia Kontostathi, Jerome Zoidakis, Eirini Baira, Lars Rönnblom, Dimitrios T. Boumpas, Prodromos Sidiropoulos, Panayotis Verginis, George Bertsias
المصدر: JCI Insight, Vol 6, Iss 21 (2021)
مصطلحات موضوعية: Autoimmunity, Inflammation, Medicine
الوصف: IL-33, a nuclear alarmin released during cell death, exerts context-specific effects on adaptive and innate immune cells, eliciting potent inflammatory responses. We screened blood, skin, and kidney tissues from patients with systemic lupus erythematosus (SLE), a systemic autoimmune disease driven by unabated type I IFN production, and found increased amounts of extracellular IL-33 complexed with neutrophil extracellular traps (NETs), correlating with severe, active disease. Using a combination of molecular, imaging, and proteomic approaches, we show that SLE neutrophils, activated by disease immunocomplexes, release IL-33–decorated NETs that stimulate robust IFN-α synthesis by plasmacytoid DCs in a manner dependent on the IL-33 receptor ST2L. IL33-silenced neutrophil-like cells cultured under lupus-inducing conditions generated NETs with diminished interferogenic effect. Importantly, NETs derived from patients with SLE are enriched in mature bioactive isoforms of IL-33 processed by the neutrophil proteases elastase and cathepsin G. Pharmacological inhibition of these proteases neutralized IL-33–dependent IFN-α production elicited by NETs. We believe these data demonstrate a novel role for cleaved IL-33 alarmin decorating NETs in human SLE, linking neutrophil activation, type I IFN production, and end-organ inflammation, with skin pathology mirroring that observed in the kidneys.
وصف الملف: electronic resource
العلاقة: https://doaj.org/toc/2379-3708Test
-
2دورية أكاديمية
المؤلفون: Marianna Naki, Olga Gourdomichali, Katerina Zonke, Fedon-Giasin Kattan, Manousos Makridakis, Georgia Kontostathi, Antonia Vlahou, Epaminondas Doxakis
المصدر: International Journal of Molecular Sciences, Vol 23, Iss 9, p 5189 (2022)
مصطلحات موضوعية: DDIT4, APEX2, proximity labeling, interactome, proteomics, LC-MS/MS, Biology (General), QH301-705.5, Chemistry, QD1-999
الوصف: DNA damage-inducible transcript 4 (DDIT4) is a ubiquitous protein whose expression is transiently increased in response to various stressors. Chronic expression has been linked to various pathologies, including neurodegeneration, inflammation, and cancer. DDIT4 is best recognized for repressing mTORC1, an essential protein complex activated by nutrients and hormones. Accordingly, DDIT4 regulates metabolism, oxidative stress, hypoxic survival, and apoptosis. Despite these well-defined biological functions, little is known about its interacting partners and their unique molecular functions. Here, fusing an enhanced ascorbate peroxidase 2 (APEX2) biotin-labeling enzyme to DDIT4 combined with mass spectrometry, the proteins in the immediate vicinity of DDIT4 in either unstressed or acute stress conditions were identified in situ. The context-dependent interacting proteomes were quantitatively but not functionally distinct. DDIT4 had twice the number of interaction partners during acute stress compared to unstressed conditions, and while the two protein lists had minimal overlap in terms of identity, the proteins’ molecular function and classification were essentially identical. Moonlighting keratins and ribosomal proteins dominated the proteomes in both unstressed and stressed conditions, with many of their members having established non-canonical and indispensable roles during stress. Multiple keratins regulate mTORC1 signaling via the recruitment of 14-3-3 proteins, whereas ribosomal proteins control translation, cell cycle progression, DNA repair, and death by sequestering critical proteins. In summary, two potentially distinct mechanisms of DDIT4 molecular function have been identified, paving the way for additional research to confirm and consolidate these findings.
وصف الملف: electronic resource
العلاقة: https://www.mdpi.com/1422-0067/23/9/5189Test; https://doaj.org/toc/1661-6596Test; https://doaj.org/toc/1422-0067Test
-
3دورية أكاديمية
المؤلفون: Olga Gourdomichali, Katerina Zonke, Fedon-Giasin Kattan, Manousos Makridakis, Georgia Kontostathi, Antonia Vlahou, Epaminondas Doxakis
المصدر: Biology, Vol 11, Iss 2, p 287 (2022)
مصطلحات موضوعية: TIA1, APEX2, proximity labeling, proteomics, LC-MS/MS, RNA binding proteins, Biology (General), QH301-705.5
الوصف: TIA1 is a broadly expressed DNA/RNA binding protein that regulates multiple aspects of RNA metabolism. It is best known for its role in stress granule assembly during the cellular stress response. Three RNA recognition motifs mediate TIA1 functions along with a prion-like domain that supports multivalent protein-protein interactions that are yet poorly characterized. Here, by fusing the enhanced ascorbate peroxidase 2 (APEX2) biotin-labeling enzyme to TIA1 combined with mass spectrometry, the proteins in the immediate vicinity of TIA1 were defined in situ. Eighty-six and 203 protein partners, mostly associated with ribonucleoprotein complexes, were identified in unstressed control and acute stress conditions, respectively. Remarkably, the repertoire of TIA1 protein partners was highly dissimilar between the two cellular states. Under unstressed control conditions, the biological processes associated with the TIA1 interactome were enriched for cytosolic ontologies related to mRNA metabolism, such as translation initiation, nucleocytoplasmic transport, and RNA catabolism, while the protein identities were primarily represented by RNA binding proteins, ribosomal subunits, and eicosanoid regulators. Under acute stress, TIA1-labeled partners displayed a broader subcellular distribution that included the chromosomes and mitochondria. The enriched biological processes included splicing, translation, and protein synthesis regulation, while the molecular function of the proteins was enriched for RNA binding activity, ribosomal subunits, DNA double-strand break repair, and amide metabolism. Altogether, these data highlight the TIA1 spatial environment with its different partners in diverse cellular states and pave the way to dissect TIA1 role in these processes.
وصف الملف: electronic resource
-
4دورية أكاديمية
مصطلحات موضوعية: biomarkers, drug targets, mass spectrometry, personalized medicine proteomics
الوصف: Clinical proteomics, the application of proteome analysis to serve a clinical purpose, represents a major field in the area of proteome research. Over 1000 manuscripts on this topic are published each year, with numbers continuously increasing. However, the anticipated outcome, the transformation of the reported findings into improvements in patient management, is not immediately evident. In this article, the value and validity of selected clinical proteomics findings are investigated, and it is assessed how far implementation has progressed. A main conclusion from this assessment is that to achieve implementation, well‐powered clinical studies are required in the appropriate population, addressing a specific clinical need and with a clear context‐of‐use. Efforts toward implementation, to be feasible, must be supported by the key players in science: publishers and funders. The authors propose a change on objectives, from additional discovery studies toward studies aiming at validation of the plethora of potential biomarkers that have been described, to demonstrate practical value of clinical proteomics. All elements required, potential biomarkers, technologies, and bio‐banked samples are available (based on today's literature), hence a change in focus from discovery toward validation and application is not only urgently necessary, but also possible based on resources available today.
العلاقة: info:eu-repo/grantAgreement/EC/H2020/752755/; https://zenodo.org/record/3358063Test; https://doi.org/10.1002/pmic.201700463Test; oai:zenodo.org:3358063
-
5دورية أكاديمية
المؤلفون: Georgia Kontostathi, Jerome Zoidakis, Manousos Makridakis, Vasiliki Lygirou, George Mermelekas, Theofilos Papadopoulos, Konstantinos Vougas, Alexios Vlamis-Gardikas, Peter Drakakis, Dimitrios Loutradis, Antonia Vlahou, Nicholas P. Anagnou, Kalliopi I. Pappa
المصدر: BioMed Research International, Vol 2017 (2017)
مصطلحات موضوعية: Medicine
الوصف: Cancer cells acquire unique secretome compositions that contribute to tumor development and metastasis. The aim of our study was to elucidate the biological processes involved in cervical cancer, by performing a proteomic analysis of the secretome from the following informative cervical cell lines: SiHa (HPV16+), HeLa (HPV18+), C33A (HPV−), and HCK1T (normal). Proteins were analyzed by 2D gel electrophoresis coupled to MALDI-TOF-MS. Enrichment of secreted proteins with characteristic profiles for each cell line was followed by the identification of differentially expressed proteins. Particularly, transforming growth factor-beta-induced protein ig-h3 (Beta ig-h3) and peroxiredoxin-2 (PRDX2) overexpression in the secretome of cancer cell lines was detected and confirmed by Western blot. Bioinformatics analysis identified the transcription factor NRF2 as a regulator of differentially expressed proteins in the cervical cancer secretome. NRF2 levels were measured by both Western blot and Multiple Reaction Monitoring (MRM) in the total cell extract of the four cell lines. NRF2 was upregulated in SiHa and C33A compared to HCK1T. In conclusion, the secreted proteins identified in cervical cancer cell lines indicate that aberrant NRF2-mediated oxidative stress response (OSR) is a prominent feature of cervical carcinogenesis.
العلاقة: http://dx.doi.org/10.1155/2017/4180703Test; https://doaj.org/toc/2314-6133Test; https://doaj.org/toc/2314-6141Test; https://doaj.org/article/76bf1d87b61741cb89ce10ad8e9ce669Test
الإتاحة: https://doi.org/10.1155/2017/4180703Test
https://doaj.org/article/76bf1d87b61741cb89ce10ad8e9ce669Test -
6
المؤلفون: Olga Gourdomichali, Katerina Zonke, Fedon-Giasin Kattan, Manousos Makridakis, Georgia Kontostathi, Antonia Vlahou, Epaminondas Doxakis
المصدر: Biology; Volume 11; Issue 2; Pages: 287
مصطلحات موضوعية: General Immunology and Microbiology, General Agricultural and Biological Sciences, TIA1, APEX2, proximity labeling, proteomics, LC-MS/MS, RNA binding proteins, stress, stress granules, General Biochemistry, Genetics and Molecular Biology
الوصف: TIA1 is a broadly expressed DNA/RNA binding protein that regulates multiple aspects of RNA metabolism. It is best known for its role in stress granule assembly during the cellular stress response. Three RNA recognition motifs mediate TIA1 functions along with a prion-like domain that supports multivalent protein-protein interactions that are yet poorly characterized. Here, by fusing the enhanced ascorbate peroxidase 2 (APEX2) biotin-labeling enzyme to TIA1 combined with mass spectrometry, the proteins in the immediate vicinity of TIA1 were defined in situ. Eighty-six and 203 protein partners, mostly associated with ribonucleoprotein complexes, were identified in unstressed control and acute stress conditions, respectively. Remarkably, the repertoire of TIA1 protein partners was highly dissimilar between the two cellular states. Under unstressed control conditions, the biological processes associated with the TIA1 interactome were enriched for cytosolic ontologies related to mRNA metabolism, such as translation initiation, nucleocytoplasmic transport, and RNA catabolism, while the protein identities were primarily represented by RNA binding proteins, ribosomal subunits, and eicosanoid regulators. Under acute stress, TIA1-labeled partners displayed a broader subcellular distribution that included the chromosomes and mitochondria. The enriched biological processes included splicing, translation, and protein synthesis regulation, while the molecular function of the proteins was enriched for RNA binding activity, ribosomal subunits, DNA double-strand break repair, and amide metabolism. Altogether, these data highlight the TIA1 spatial environment with its different partners in diverse cellular states and pave the way to dissect TIA1 role in these processes.
وصف الملف: application/pdf
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4bef9b9b8ea1b1ea77439d24aa7ac2bfTest
https://pubmed.ncbi.nlm.nih.gov/35205152Test -
7
المؤلفون: Àngel Argilés, Vasiliki Lygirou, Eleni Petra, Flore Duranton, Harald Mischak, Manousos Makridakis, Jerome Zoidakis, Szymon Filip, Antonia Vlahou, Georgia Kontostathi, Rafael Stroggilos
المصدر: Scientific Reports, Vol 10, Iss 1, Pp 1-12 (2020)
Scientific Reportsمصطلحات موضوعية: Male, 0301 basic medicine, Oncology, 030232 urology & nephrology, lcsh:Medicine, Disease, Mass Spectrometry, Cohort Studies, Prognostic markers, 0302 clinical medicine, Multiplex, Longitudinal Studies, lcsh:Science, Aged, 80 and over, Multidisciplinary, Immunoglobulin heavy constant alpha 1, Middle Aged, Prognosis, Blood proteins, Hemoglobin Subunits, Disease Progression, Female, medicine.symptom, Complement C1 Inhibitor Protein, Glomerular Filtration Rate, Adult, medicine.medical_specialty, Quantitative proteomics, Proteomic analysis, Renal function, Article, 03 medical and health sciences, Internal medicine, Alpha-Globulins, medicine, Humans, Renal Insufficiency, Chronic, Aged, business.industry, lcsh:R, medicine.disease, 030104 developmental biology, Albuminuria, Muramidase, lcsh:Q, beta 2-Microglobulin, business, Biomarkers, Follow-Up Studies, Kidney disease
الوصف: Current diagnostic measures for Chronic Kidney Disease (CKD) include detection of reduced estimated glomerular filtration rate (eGFR) and albuminuria, which have suboptimal accuracies in predicting disease progression. The disease complexity and heterogeneity underscore the need for multiplex quantification of different markers. The goal of this study was to determine the association of six previously reported CKD-associated plasma proteins [B2M (Beta-2-microglobulin), SERPINF1 (Pigment epithelium-derived factor), AMBP (Protein AMBP), LYZ (Lysozyme C), HBB (Hemoglobin subunit beta) and IGHA1 (Immunoglobulin heavy constant alpha 1)], as measured in a multiplex format, with kidney function, and outcome. Antibody-free, multiple reaction monitoring mass spectrometry (MRM) assays were developed, characterized for their analytical performance, and used for the analysis of 72 plasma samples from a patient cohort with longitudinal follow-up. The MRM significantly correlated (Rho = 0.5–0.9) with results from respective ELISA. Five proteins [AMBP, B2M, LYZ, HBB and SERPINF1] were significantly associated with eGFR, with the three former also associated with unfavorable outcome. The combination of these markers provided stronger associations with outcome (p
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f45ed4d3b1c70609a58b348a2d897aa9Test
http://link.springer.com/article/10.1038/s41598-020-61496-zTest -
8
المؤلفون: Katerina Gkirtzimanaki, Hariklia Gakiopoulou, Maija-Leena Eloranta, Manousos Makridakis, Georgia Kontostathi, Garyfalia Papadaki, Eirini Baira, Panayotis Verginis, Prodromos Sidiropoulos, Dimitrios T. Boumpas, Lars Rönnblom, Jerome Zoidakis, George Bertsias, Spiros Georgakis, Elias Drakos
المصدر: JCI Insight
مصطلحات موضوعية: Neutrophils, Lupus, Inflammation, Autoimmunity, Cathepsin G, medicine.disease_cause, Extracellular Traps, chemistry.chemical_compound, Interferon, medicine, Humans, Lupus Erythematosus, Systemic, Rheumatology and Autoimmunity, Reumatologi och inflammation, Systemic lupus erythematosus, Innate immune system, Elastase, Interferon-alpha, Dendritic Cells, General Medicine, Neutrophil extracellular traps, Interleukin-33, medicine.disease, chemistry, Case-Control Studies, Immunology, Cytokines, medicine.symptom, Research Article, medicine.drug
الوصف: Interleukin-33 (IL-33), a nuclear alarmin released during cell death, exerts context-specific effects on adaptive and innate immune cells eliciting potent inflammatory responses. We screened blood, skin and kidney tissues from patients with Systemic Lupus Erythematosus (SLE), a systemic autoimmune disease driven by unabated type I interferon (IFN) production, and found increased amounts of extracellular IL-33 complexed with Neutrophil Extracellular Traps (NETs), correlating with severe, active disease. Using a combination of molecular, imaging and proteomic approaches, we show that SLE neutrophils -activated by disease immunocomplexes- release IL-33-decorated NETs that stimulate robust IFNα synthesis by plasmacytoid dendritic cells (pDCs) in an IL-33-receptor (ST2L)-dependent manner. IL33-silenced neutrophil-like cells cultured under lupus-inducing conditions generated NETs with diminished interferogenic effect. Importantly, SLE patient-derived NETs are enriched in mature bioactive isoforms of IL-33 processed by the neutrophil proteases elastase and cathepsin G. Pharmacological inhibition of these proteases neutralized IL-33-dependent IFNα production elicited by NETs. These data demonstrate a novel role for cleaved IL-33 alarmin decorating NETs in human SLE, linking neutrophil activation, type I IFN production and end-organ inflammation with skin pathology mirroring that observed in the kidneys.
وصف الملف: application/pdf
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4f910330fcfe8059d38c8071422aaa4eTest
http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-464303Test -
9
المؤلفون: Antonios Sfakianakis, Vasiliki Lygirou, Konstantinos Vougas, George Daskalakis, Jerome Zoidakis, Kalliopi I. Pappa, Georgia Kontostathi, Nicholas P. Anagnou, Manousos Makridakis
المصدر: Oncology Reports.
مصطلحات موضوعية: 0301 basic medicine, Cervical cancer, Cancer Research, Oncogene, biology, Cancer, General Medicine, Cell cycle, medicine.disease, biology.organism_classification, medicine.disease_cause, Molecular medicine, HeLa, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, Oncology, 030220 oncology & carcinogenesis, Proteome, Cancer research, medicine, Carcinogenesis
الوصف: Cervical cancer remains the fourth most common and most lethal type of cancer in women, despite the applied regular screening and prevention strategies, while the available treatment schemes still pose a threat to fertility. Substantial understanding of the underlying mechanisms and development of novel diagnostic, prognostic and therapeutic approaches are critical steps for improving cervical cancer management. Towards this goal, a comparative proteomic analysis was conducted between three cervical cancer cell lines (HeLa: HPV18+, SiHa: HPV16+, C33A: HPV‑) and normal cervical keratinocytes (HCK1T). The total cell extract of each cell line was analyzed by liquid chromatography coupled to tandem mass spectrometry (LC‑MS/MS). Differential expression analysis revealed 919, 826 and 1,370 deregulated proteins in the comparisons of HeLa, SiHa and C33A with HCK1T cell lines, respectively. Pathway enrichment analysis of the differentially expressed proteins highlighted common cancer characteristics such as high metabolic demands and increased cell turnover, confirming the validity of the proteomic results. Extensive literature mining of the consistently differentially expressed proteins that resulted from the three comparisons was performed leading to a shortlist of 21 proteins that are potentially involved in cervical malignancy. The criteria for this shortlisting were the association of the proteins with various types of cancer, while there is no study as yet associating their expression to cervical cancer. Moreover, the expression trend of two of the shortlisted proteins was validated using western blot analysis. The proteomic datasets generated in this study can be utilized to enrich the current knowledge on cervical cancer pathology and unveil key molecular mechanisms of carcinogenesis. In conclusion, the shortlist of consistently deregulated proteins between cervical cancer cell lines and normal cervical keratinocytes can be used for validation in clinical samples and in functional investigation experiments that could ultimately lead to the discovery of novel disease biomarkers and drug targets.
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::11cdb98b5953d001a0f12d76c4d46fc1Test
https://doi.org/10.3892/or.2019.7269Test -
10
المؤلفون: George Mermelekas, Peter Drakakis, Kalliopi I. Pappa, Antonia Vlahou, Konstantinos Vougas, Jerome Zoidakis, Georgia Kontostathi, Theofilos Papadopoulos, Vasiliki Lygirou, Nicholas P. Anagnou, Alexios Vlamis-Gardikas, Dimitrios Loutradis, Manousos Makridakis
المصدر: BioMed Research International, Vol 2017 (2017)
BioMed Research Internationalمصطلحات موضوعية: Proteomics, 0301 basic medicine, Article Subject, Carcinogenesis, NF-E2-Related Factor 2, Uterine Cervical Neoplasms, lcsh:Medicine, Peroxiredoxin 2, medicine.disease_cause, General Biochemistry, Genetics and Molecular Biology, Metastasis, HeLa, 03 medical and health sciences, Western blot, Tandem Mass Spectrometry, Transforming Growth Factor beta, Cell Line, Tumor, medicine, Humans, Electrophoresis, Gel, Two-Dimensional, Extracellular Matrix Proteins, Human papillomavirus 16, General Immunology and Microbiology, medicine.diagnostic_test, biology, Papillomavirus Infections, lcsh:R, Computational Biology, Peroxiredoxins, General Medicine, Transforming growth factor beta, medicine.disease, biology.organism_classification, Cell biology, Gene Expression Regulation, Neoplastic, 030104 developmental biology, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Cancer cell, Cancer research, biology.protein, Female, Peptides, Algorithms, HeLa Cells, Signal Transduction, Research Article
الوصف: Cancer cells acquire unique secretome compositions that contribute to tumor development and metastasis. The aim of our study was to elucidate the biological processes involved in cervical cancer, by performing a proteomic analysis of the secretome from the following informative cervical cell lines: SiHa (HPV16+), HeLa (HPV18+), C33A (HPV−), and HCK1T (normal). Proteins were analyzed by 2D gel electrophoresis coupled to MALDI-TOF-MS. Enrichment of secreted proteins with characteristic profiles for each cell line was followed by the identification of differentially expressed proteins. Particularly, transforming growth factor-beta-induced protein ig-h3 (Beta ig-h3) and peroxiredoxin-2 (PRDX2) overexpression in the secretome of cancer cell lines was detected and confirmed by Western blot. Bioinformatics analysis identified the transcription factor NRF2 as a regulator of differentially expressed proteins in the cervical cancer secretome. NRF2 levels were measured by both Western blot and Multiple Reaction Monitoring (MRM) in the total cell extract of the four cell lines. NRF2 was upregulated in SiHa and C33A compared to HCK1T. In conclusion, the secreted proteins identified in cervical cancer cell lines indicate that aberrant NRF2-mediated oxidative stress response (OSR) is a prominent feature of cervical carcinogenesis.
وصف الملف: text/xhtml
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5cf882f01f3e5da8493ab44bf0a9b9b2Test
https://doi.org/10.1155/2017/4180703Test
النص الكامل