Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology
Abstract Background Neurological disorders are the second leading cause of death and the leading cause of disability in the world. Thus, the development of novel disease-modifying strategies is clearly warranted. We have previously developed a therapeutic approach using mouse targeted rabies virus g...
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BMC
2025-05-01
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| Series: | Translational Neurodegeneration |
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| Online Access: | https://doi.org/10.1186/s40035-025-00484-7 |
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| author | Maria Izco Carlos Sola Martin Schleef Marco Schmeer María de Toro Guglielmo Verona Estefania Carlos Alejandro Reinares-Sebastian Sandra Colina Maria Eugenia Marzo-Sola Josune Garcia-Sanmartin Joaquín Fernández-Irigoyen Enrique Santamaría Rodolfo Mugica-Vidal Javier Blesa Lydia Alvarez-Erviti |
| author_facet | Maria Izco Carlos Sola Martin Schleef Marco Schmeer María de Toro Guglielmo Verona Estefania Carlos Alejandro Reinares-Sebastian Sandra Colina Maria Eugenia Marzo-Sola Josune Garcia-Sanmartin Joaquín Fernández-Irigoyen Enrique Santamaría Rodolfo Mugica-Vidal Javier Blesa Lydia Alvarez-Erviti |
| author_sort | Maria Izco |
| collection | DOAJ |
| description | Abstract Background Neurological disorders are the second leading cause of death and the leading cause of disability in the world. Thus, the development of novel disease-modifying strategies is clearly warranted. We have previously developed a therapeutic approach using mouse targeted rabies virus glycoprotein (RVG) extracellular vesicles (EVs) to deliver minicircles (MCs) expressing shRNA (shRNA-MCs) to induce long-term α-synuclein down-regulation. Although the previous therapy successfully reduced the pathology, the clinical translation was extremely unlikely since they were mouse extracellular vesicles. Methods To overcome this limitation, we developed a source of human RVG-EVs compatible with a personalized therapy using immature dendritic cells. Human peripheral blood monocytes were differentiated in vitro into immature dendritic cells, which were transfected to express the RVG peptide. RVG-EVs containing shRNA-MCs, loaded by electroporation, were injected intravenously in the α-synuclein performed fibril (PFF) mouse model. Level of α-synuclein, phosphorylated α-synuclein aggregates, dopaminergic neurons and motor function were evaluated 90 days after the treatment. To confirm that EVs derived from patients were suitable as a vehicle, proteomic analysis of EVs derived from control, initial and advanced Parkinson’s disease was performed. Results The shRNA-MCs could be successfully loaded into human RVG-EVs and downregulate α-synuclein in SH-SY5Y cells. Intravenous injection of the shRNA-MC-loaded RVG-EVs induced long-term downregulation of α-synuclein mRNA expression and protein level, decreased α-synuclein aggregates, prevented dopaminergic cell death and ameliorated motor impairment in the α-synuclein PFF mouse model. Moreover, we confirmed that the EVs from PD patients are suitable as a personalized therapeutic vehicle. Conclusion Our study confirmed the therapeutic potential of shRNA-MCs delivered by human RVG-EVs for long-term treatment of neurodegenerative diseases. These results pave the way for clinical use of this approach. |
| format | Article |
| id | doaj-art-c58c7dc733d94a919f64b676b3f20f92 |
| institution | OA Journals |
| issn | 2047-9158 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | BMC |
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| series | Translational Neurodegeneration |
| spelling | doaj-art-c58c7dc733d94a919f64b676b3f20f922025-08-20T02:03:32ZengBMCTranslational Neurodegeneration2047-91582025-05-0114111710.1186/s40035-025-00484-7Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathologyMaria Izco0Carlos Sola1Martin Schleef2Marco Schmeer3María de Toro4Guglielmo Verona5Estefania Carlos6Alejandro Reinares-Sebastian7Sandra Colina8Maria Eugenia Marzo-Sola9Josune Garcia-Sanmartin10Joaquín Fernández-Irigoyen11Enrique Santamaría12Rodolfo Mugica-Vidal13Javier Blesa14Lydia Alvarez-Erviti15Laboratory of Molecular Neurobiology, Center for Biomedical Research of La Rioja (CIBIR)Transfusion Center and Blood Bank of La RiojaPlasmidFactory GmbHPlasmidFactory GmbHGenomics and Bioinformatics Core Facility, Center for Biomedical Research of La Rioja (CIBIR)Centre for Amyloidosis, UCL Medical SchoolLaboratory of Molecular Neurobiology, Center for Biomedical Research of La Rioja (CIBIR)HM CINAC (Centro Integral de Neurociencias Abarca Campal), Hospital Universitario HM Puerta del Sur, HM HospitalesServicio de Neurología, Hospital San PedroServicio de Neurología, Hospital San PedroAngiogenesis Group, Oncology Area, Center for Biomedical Research of La Rioja (CIBIR)Clinical Neuroproteomics Unit, Proteomics Platform, Navarrabiomed, Hospitalario, Universitario de Navarra (HUN)Clinical Neuroproteomics Unit, Proteomics Platform, Navarrabiomed, Hospitalario, Universitario de Navarra (HUN)Department of Mechanical Engineering, University of La RiojaHM CINAC (Centro Integral de Neurociencias Abarca Campal), Hospital Universitario HM Puerta del Sur, HM HospitalesLaboratory of Molecular Neurobiology, Center for Biomedical Research of La Rioja (CIBIR)Abstract Background Neurological disorders are the second leading cause of death and the leading cause of disability in the world. Thus, the development of novel disease-modifying strategies is clearly warranted. We have previously developed a therapeutic approach using mouse targeted rabies virus glycoprotein (RVG) extracellular vesicles (EVs) to deliver minicircles (MCs) expressing shRNA (shRNA-MCs) to induce long-term α-synuclein down-regulation. Although the previous therapy successfully reduced the pathology, the clinical translation was extremely unlikely since they were mouse extracellular vesicles. Methods To overcome this limitation, we developed a source of human RVG-EVs compatible with a personalized therapy using immature dendritic cells. Human peripheral blood monocytes were differentiated in vitro into immature dendritic cells, which were transfected to express the RVG peptide. RVG-EVs containing shRNA-MCs, loaded by electroporation, were injected intravenously in the α-synuclein performed fibril (PFF) mouse model. Level of α-synuclein, phosphorylated α-synuclein aggregates, dopaminergic neurons and motor function were evaluated 90 days after the treatment. To confirm that EVs derived from patients were suitable as a vehicle, proteomic analysis of EVs derived from control, initial and advanced Parkinson’s disease was performed. Results The shRNA-MCs could be successfully loaded into human RVG-EVs and downregulate α-synuclein in SH-SY5Y cells. Intravenous injection of the shRNA-MC-loaded RVG-EVs induced long-term downregulation of α-synuclein mRNA expression and protein level, decreased α-synuclein aggregates, prevented dopaminergic cell death and ameliorated motor impairment in the α-synuclein PFF mouse model. Moreover, we confirmed that the EVs from PD patients are suitable as a personalized therapeutic vehicle. Conclusion Our study confirmed the therapeutic potential of shRNA-MCs delivered by human RVG-EVs for long-term treatment of neurodegenerative diseases. These results pave the way for clinical use of this approach.https://doi.org/10.1186/s40035-025-00484-7Parkinson’s diseaseGene therapyα-SynucleinHuman targeted extracellular vesiclesShRNA minicircles |
| spellingShingle | Maria Izco Carlos Sola Martin Schleef Marco Schmeer María de Toro Guglielmo Verona Estefania Carlos Alejandro Reinares-Sebastian Sandra Colina Maria Eugenia Marzo-Sola Josune Garcia-Sanmartin Joaquín Fernández-Irigoyen Enrique Santamaría Rodolfo Mugica-Vidal Javier Blesa Lydia Alvarez-Erviti Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology Translational Neurodegeneration Parkinson’s disease Gene therapy α-Synuclein Human targeted extracellular vesicles ShRNA minicircles |
| title | Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology |
| title_full | Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology |
| title_fullStr | Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology |
| title_full_unstemmed | Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology |
| title_short | Development of human targeted extracellular vesicles loaded with shRNA minicircles to prevent parkinsonian pathology |
| title_sort | development of human targeted extracellular vesicles loaded with shrna minicircles to prevent parkinsonian pathology |
| topic | Parkinson’s disease Gene therapy α-Synuclein Human targeted extracellular vesicles ShRNA minicircles |
| url | https://doi.org/10.1186/s40035-025-00484-7 |
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