Publicação
Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease
| Resumo: | Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease, presenting a wide spectrum of metabolic dysregulation. However, its progression extends beyond the liver, affecting extrahepatic organs. In skeletal muscle, sarcopenia and myosteatosis are often associated with advanced MASLD. Previous studies showed that microRNA-21 (miR-21) continuously increases with MASLD progression in the liver, serum and skeletal muscle, while its ablation improves outcomes in animal models. In this work, we aimed to evaluate whether liver-derived extracellular vesicles (EVs) containing miR-21 contribute to skeletal muscle dysfunction in MASLD, using both a diet-induced MASLD mouse model and an in vitro model of hepatocyte-to-myotube communication. Mir21 whole-body knockout (WB-KO); Mir21 Alb-Cre KO (Alb-Cre-KO) and wildtype (WT) mice were fed a high-fat diet (HFD), with high fructose/glucose drinking water for 20 weeks or a standard diet. AML12-derived EVs were incubated with C2C12 myotubes for miR-21 assays and gene expression was evaluated by RT-qPCR. HFD mice displayed hepatic steatosis without muscle atrophy or changes in miR-21 levels. However, WB-KO mice showed increased levels of muscle regeneration factors Pax7 and Myod1, counteracting the decreases observed in WT HFD-fed mice. Additionally, Alb-Cre-KO HFD-fed mice exhibited reduced expression of atrophy-related gene Fbxo32 compared to WT HFD-fed mice. Hepatocyte miR-21 overexpression led to an increase in miR-21- loaded EVs, which subsequently elevated miR-21 expression in myotubes. This resulted in increased levels of genes associated with lipid metabolism, inflammation, muscle regeneration and atrophy, while decreasing levels of muscle growth inhibitor Myostatin. Notably, miR-21 inhibition in palmitic acidtreated hepatocytes reduced secreted miR-21 in EVs. However, myotube miR-21 expression remained unchanged after incubation. Overall, these findings suggest that miR-21 ablation promotes skeletal muscle regeneration and inhibits atrophy. The in vitro model highlighted the role of hepatocytes-derived miR-21-loaded EVs in inducing myotube dysfunction, closely resembling transcription profiles seen in MASLD-associated sarcopenia patients. |
|---|---|
| Autores principais: | Rodrigues, José Alberto Correia Pereira |
| Assunto: | Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| Ano: | 2025 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso embargado |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| _version_ | 1866811283814219776 |
|---|---|
| author | Rodrigues, José Alberto Correia Pereira |
| author_facet | Rodrigues, José Alberto Correia Pereira |
| author_role | author |
| contributor_name_str_mv | Simão, André Daniel Lopes, 1990- Rodrigues, Maria Gabriela, 1965- Repositório Científico de Acesso Aberto da ULisboa |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Rodrigues, José Alberto Correia Pereira\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | Simão, André Daniel Lopes, 1990- Rodrigues, Maria Gabriela, 1965- Repositório Científico de Acesso Aberto da ULisboa |
| datacite.creators.creator.creatorName.fl_str_mv | Rodrigues, José Alberto Correia Pereira |
| datacite.date.Accepted.fl_str_mv | 2025-01-01T00:00:00Z |
| datacite.date.available.fl_str_mv | 2027-02-20T00:00:00Z |
| datacite.date.embargoed.fl_str_mv | 2027-02-20T00:00:00Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_f1cf |
| datacite.subjects.subject.fl_str_mv | Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| datacite.titles.title.fl_str_mv | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| dc.contributor.none.fl_str_mv | Simão, André Daniel Lopes, 1990- Rodrigues, Maria Gabriela, 1965- Repositório Científico de Acesso Aberto da ULisboa |
| dc.creator.none.fl_str_mv | Rodrigues, José Alberto Correia Pereira |
| dc.date.Accepted.fl_str_mv | 2025-01-01T00:00:00Z |
| dc.date.available.fl_str_mv | 2027-02-20T00:00:00Z |
| dc.date.embargoed.fl_str_mv | 2027-02-20T00:00:00Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10400.5/97781 |
| dc.language.none.fl_str_mv | eng |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_f1cf |
| dc.subject.none.fl_str_mv | Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| dc.title.fl_str_mv | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_bdcc |
| description | Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease, presenting a wide spectrum of metabolic dysregulation. However, its progression extends beyond the liver, affecting extrahepatic organs. In skeletal muscle, sarcopenia and myosteatosis are often associated with advanced MASLD. Previous studies showed that microRNA-21 (miR-21) continuously increases with MASLD progression in the liver, serum and skeletal muscle, while its ablation improves outcomes in animal models. In this work, we aimed to evaluate whether liver-derived extracellular vesicles (EVs) containing miR-21 contribute to skeletal muscle dysfunction in MASLD, using both a diet-induced MASLD mouse model and an in vitro model of hepatocyte-to-myotube communication. Mir21 whole-body knockout (WB-KO); Mir21 Alb-Cre KO (Alb-Cre-KO) and wildtype (WT) mice were fed a high-fat diet (HFD), with high fructose/glucose drinking water for 20 weeks or a standard diet. AML12-derived EVs were incubated with C2C12 myotubes for miR-21 assays and gene expression was evaluated by RT-qPCR. HFD mice displayed hepatic steatosis without muscle atrophy or changes in miR-21 levels. However, WB-KO mice showed increased levels of muscle regeneration factors Pax7 and Myod1, counteracting the decreases observed in WT HFD-fed mice. Additionally, Alb-Cre-KO HFD-fed mice exhibited reduced expression of atrophy-related gene Fbxo32 compared to WT HFD-fed mice. Hepatocyte miR-21 overexpression led to an increase in miR-21- loaded EVs, which subsequently elevated miR-21 expression in myotubes. This resulted in increased levels of genes associated with lipid metabolism, inflammation, muscle regeneration and atrophy, while decreasing levels of muscle growth inhibitor Myostatin. Notably, miR-21 inhibition in palmitic acidtreated hepatocytes reduced secreted miR-21 in EVs. However, myotube miR-21 expression remained unchanged after incubation. Overall, these findings suggest that miR-21 ablation promotes skeletal muscle regeneration and inhibits atrophy. The in vitro model highlighted the role of hepatocytes-derived miR-21-loaded EVs in inducing myotube dysfunction, closely resembling transcription profiles seen in MASLD-associated sarcopenia patients. |
| dirty | 0 |
| eu_rights_str_mv | embargoedAccess |
| format | masterThesis |
| fulltext.url.fl_str_mv | https://repositorio.ulisboa.pt/bitstreams/dce5473a-8cfb-4fab-91fc-50e3cba8cb61/download |
| id | ul_8fd4729b48e400a1d3cf5d01ed0bafb2 |
| identifier.url.fl_str_mv | http://hdl.handle.net/10400.5/97781 |
| instacron_str | ul |
| institution | Universidade de Lisboa |
| instname_str | Universidade de Lisboa |
| language | eng |
| network_acronym_str | ul |
| network_name_str | Repositório da Universidade de Lisboa |
| oai_identifier_str | oai:repositorio.ulisboa.pt:10400.5/97781 |
| organization_str_mv | urn:organizationAcronym:ul |
| person_str_mv | Rodrigues, José Alberto Correia Pereira |
| publishDate | 2025 |
| reponame_str | Repositório da Universidade de Lisboa |
| repository_id_str | urn:repositoryAcronym:ul |
| service_str_mv | urn:repositoryAcronym:ul |
| spelling | engpt_PTMetabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease, presenting a wide spectrum of metabolic dysregulation. However, its progression extends beyond the liver, affecting extrahepatic organs. In skeletal muscle, sarcopenia and myosteatosis are often associated with advanced MASLD. Previous studies showed that microRNA-21 (miR-21) continuously increases with MASLD progression in the liver, serum and skeletal muscle, while its ablation improves outcomes in animal models. In this work, we aimed to evaluate whether liver-derived extracellular vesicles (EVs) containing miR-21 contribute to skeletal muscle dysfunction in MASLD, using both a diet-induced MASLD mouse model and an in vitro model of hepatocyte-to-myotube communication. Mir21 whole-body knockout (WB-KO); Mir21 Alb-Cre KO (Alb-Cre-KO) and wildtype (WT) mice were fed a high-fat diet (HFD), with high fructose/glucose drinking water for 20 weeks or a standard diet. AML12-derived EVs were incubated with C2C12 myotubes for miR-21 assays and gene expression was evaluated by RT-qPCR. HFD mice displayed hepatic steatosis without muscle atrophy or changes in miR-21 levels. However, WB-KO mice showed increased levels of muscle regeneration factors Pax7 and Myod1, counteracting the decreases observed in WT HFD-fed mice. Additionally, Alb-Cre-KO HFD-fed mice exhibited reduced expression of atrophy-related gene Fbxo32 compared to WT HFD-fed mice. Hepatocyte miR-21 overexpression led to an increase in miR-21- loaded EVs, which subsequently elevated miR-21 expression in myotubes. This resulted in increased levels of genes associated with lipid metabolism, inflammation, muscle regeneration and atrophy, while decreasing levels of muscle growth inhibitor Myostatin. Notably, miR-21 inhibition in palmitic acidtreated hepatocytes reduced secreted miR-21 in EVs. However, myotube miR-21 expression remained unchanged after incubation. Overall, these findings suggest that miR-21 ablation promotes skeletal muscle regeneration and inhibits atrophy. The in vitro model highlighted the role of hepatocytes-derived miR-21-loaded EVs in inducing myotube dysfunction, closely resembling transcription profiles seen in MASLD-associated sarcopenia patients.application/pdfpt_PTRole of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver DiseaseRodrigues, José Alberto Correia PereiraSimão, André Daniel Lopes, 1990-Rodrigues, Maria Gabriela, 1965-HostingInstitutionOrganizationalRepositório Científico de Acesso Aberto da ULisboae-mailmailto:repositorio@reitoria.ulisboa.ptrepositorio@reitoria.ulisboa.pt202520242027-02-20T00:00:00Z2025-01-01T00:00:00ZHandlehttp://hdl.handle.net/10400.5/97781http://purl.org/coar/access_right/c_f1cfembargoed accessComunicação fígado para músculoMASLDmiR-21SarcopeniaVesículas extracelularesTeses de mestrado - 20252071976 bytesliteraturehttp://purl.org/coar/resource_type/c_bdccmaster thesishttp://purl.org/coar/access_right/c_f1cfapplication/pdffulltexthttps://repositorio.ulisboa.pt/bitstreams/dce5473a-8cfb-4fab-91fc-50e3cba8cb61/download |
| spellingShingle | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease Rodrigues, José Alberto Correia Pereira Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| status | SINGLETON |
| subject.fl_str_mv | Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| title | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| title_full | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| title_fullStr | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| title_full_unstemmed | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| title_short | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| title_sort | Role of microRNA-21-loaded extracellular vesicles in liver-muscle intercommunication during Metabolic Dysfunction–associated Steatotic Liver Disease |
| topic | Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| topic_facet | Comunicação fígado para músculo MASLD miR-21 Sarcopenia Vesículas extracelulares Teses de mestrado - 2025 |
| url | http://hdl.handle.net/10400.5/97781 |
| visible | 1 |