Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.3390/ijms23169025 http://hdl.handle.net/11449/240705 |
Resumo: | Advanced quantitative bioanalytical approaches in combination with network analyses allow us to answer complex biological questions, such as the description of changes in protein profiles under disease conditions or upon treatment with drugs. In the present work, three quantitative proteomic approaches—either based on labelling or not—in combination with network analyses were applied to a new in vitro cellular model of nonalcoholic fatty liver disease (NAFLD) for the first time. This disease is characterized by the accumulation of lipids, inflammation, fibrosis, and insulin resistance. Hepatic G2 cells were used as model, and NAFLD was induced by a complex of oleic acid and bovine albumin. The development of the disease was verified by lipid vesicle staining and by the increase in the expression of perilipin-2—a protein constitutively present in the vesicles during NAFLD. The nLC–MS/MS analyses of peptide samples obtained from three different proteomic approaches resulted in accurate and reproducible quantitative data of protein fold-change expressed in NAFLD versus control cells. The differentially regulated proteins were used to evaluate the involved and statistically enriched pathways. Network analyses highlighted several functional and disease modules affected by NAFLD, such as inflammation, oxidative stress defense, cell proliferation, and ferroptosis. Each quantitative approach allowed the identification of similar modulated pathways. The combination of the three approaches improved the power of statistical network analyses by increasing the number of involved proteins and their fold-change. In conclusion, the application of advanced bioanalytical approaches in combination with pathway analyses allows the in-depth and accurate description of the protein profile of an in vitro cellular model of NAFLD by using high-resolution quantitative mass spectrometry data. This model could be extremely useful in the discovery of new drugs to modulate the equilibrium NAFLD health state. |
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Repositório Institucional da UNESP |
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Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical ApproachesNAFLDnano liquid chromatographyproteinssignalingtandem mass spectrometryAdvanced quantitative bioanalytical approaches in combination with network analyses allow us to answer complex biological questions, such as the description of changes in protein profiles under disease conditions or upon treatment with drugs. In the present work, three quantitative proteomic approaches—either based on labelling or not—in combination with network analyses were applied to a new in vitro cellular model of nonalcoholic fatty liver disease (NAFLD) for the first time. This disease is characterized by the accumulation of lipids, inflammation, fibrosis, and insulin resistance. Hepatic G2 cells were used as model, and NAFLD was induced by a complex of oleic acid and bovine albumin. The development of the disease was verified by lipid vesicle staining and by the increase in the expression of perilipin-2—a protein constitutively present in the vesicles during NAFLD. The nLC–MS/MS analyses of peptide samples obtained from three different proteomic approaches resulted in accurate and reproducible quantitative data of protein fold-change expressed in NAFLD versus control cells. The differentially regulated proteins were used to evaluate the involved and statistically enriched pathways. Network analyses highlighted several functional and disease modules affected by NAFLD, such as inflammation, oxidative stress defense, cell proliferation, and ferroptosis. Each quantitative approach allowed the identification of similar modulated pathways. The combination of the three approaches improved the power of statistical network analyses by increasing the number of involved proteins and their fold-change. In conclusion, the application of advanced bioanalytical approaches in combination with pathway analyses allows the in-depth and accurate description of the protein profile of an in vitro cellular model of NAFLD by using high-resolution quantitative mass spectrometry data. This model could be extremely useful in the discovery of new drugs to modulate the equilibrium NAFLD health state.Department of Pharmaceutical Sciences University of Milan, Via L. Mangiagalli 25Department of Human Science and Quality of Life Promotion Telematic University San RaffaeleMedical School Sao Paulo State UniversityUnitech OMICs Platform University of Milan, Viale Ortles 22/4Medical School Sao Paulo State UniversityUniversity of MilanTelematic University San RaffaeleUniversidade Estadual Paulista (UNESP)Altomare, Alessandra AnnaAiello, GildaGarcia, Jessica Leite [UNESP]Garrone, GiuliaZoanni, BeatriceCarini, MarinaAldini, GiancarloD’Amato, Alfonsina2023-03-01T20:29:16Z2023-03-01T20:29:16Z2022-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.3390/ijms23169025International Journal of Molecular Sciences, v. 23, n. 16, 2022.1422-00671661-6596http://hdl.handle.net/11449/24070510.3390/ijms231690252-s2.0-85136617059Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengInternational Journal of Molecular Sciencesinfo:eu-repo/semantics/openAccess2023-03-01T20:29:17Zoai:repositorio.unesp.br:11449/240705Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-03-01T20:29:17Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
spellingShingle |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches Altomare, Alessandra Anna NAFLD nano liquid chromatography proteins signaling tandem mass spectrometry |
title_short |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_full |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_fullStr |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_full_unstemmed |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_sort |
Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
author |
Altomare, Alessandra Anna |
author_facet |
Altomare, Alessandra Anna Aiello, Gilda Garcia, Jessica Leite [UNESP] Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina |
author_role |
author |
author2 |
Aiello, Gilda Garcia, Jessica Leite [UNESP] Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
University of Milan Telematic University San Raffaele Universidade Estadual Paulista (UNESP) |
dc.contributor.author.fl_str_mv |
Altomare, Alessandra Anna Aiello, Gilda Garcia, Jessica Leite [UNESP] Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina |
dc.subject.por.fl_str_mv |
NAFLD nano liquid chromatography proteins signaling tandem mass spectrometry |
topic |
NAFLD nano liquid chromatography proteins signaling tandem mass spectrometry |
description |
Advanced quantitative bioanalytical approaches in combination with network analyses allow us to answer complex biological questions, such as the description of changes in protein profiles under disease conditions or upon treatment with drugs. In the present work, three quantitative proteomic approaches—either based on labelling or not—in combination with network analyses were applied to a new in vitro cellular model of nonalcoholic fatty liver disease (NAFLD) for the first time. This disease is characterized by the accumulation of lipids, inflammation, fibrosis, and insulin resistance. Hepatic G2 cells were used as model, and NAFLD was induced by a complex of oleic acid and bovine albumin. The development of the disease was verified by lipid vesicle staining and by the increase in the expression of perilipin-2—a protein constitutively present in the vesicles during NAFLD. The nLC–MS/MS analyses of peptide samples obtained from three different proteomic approaches resulted in accurate and reproducible quantitative data of protein fold-change expressed in NAFLD versus control cells. The differentially regulated proteins were used to evaluate the involved and statistically enriched pathways. Network analyses highlighted several functional and disease modules affected by NAFLD, such as inflammation, oxidative stress defense, cell proliferation, and ferroptosis. Each quantitative approach allowed the identification of similar modulated pathways. The combination of the three approaches improved the power of statistical network analyses by increasing the number of involved proteins and their fold-change. In conclusion, the application of advanced bioanalytical approaches in combination with pathway analyses allows the in-depth and accurate description of the protein profile of an in vitro cellular model of NAFLD by using high-resolution quantitative mass spectrometry data. This model could be extremely useful in the discovery of new drugs to modulate the equilibrium NAFLD health state. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-08-01 2023-03-01T20:29:16Z 2023-03-01T20:29:16Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.3390/ijms23169025 International Journal of Molecular Sciences, v. 23, n. 16, 2022. 1422-0067 1661-6596 http://hdl.handle.net/11449/240705 10.3390/ijms23169025 2-s2.0-85136617059 |
url |
http://dx.doi.org/10.3390/ijms23169025 http://hdl.handle.net/11449/240705 |
identifier_str_mv |
International Journal of Molecular Sciences, v. 23, n. 16, 2022. 1422-0067 1661-6596 10.3390/ijms23169025 2-s2.0-85136617059 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
International Journal of Molecular Sciences |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1803046860401672192 |