A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model
Abstract Background Alpha-synuclein (αSyn) is a major player in the pathophysiology of synucleinopathies, which include Parkinson’s disease, dementia with Lewy bodies, and multiple system atrophy. To date, there is no disease-modifying therapy available for these synucleinopathies. Furthermore, the...
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BMC
2025-06-01
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| Series: | Translational Neurodegeneration |
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| Online Access: | https://doi.org/10.1186/s40035-025-00486-5 |
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| author | Matthias Höllerhage Linghan Duan Oscar Wing Ho Chua Claudia Moebius Svenja H. Bothe Kristina Losse Rebecca Kotzur Kristina Lau Franziska Hopfner Franziska Richter Christian Wahl-Schott Marc Bickle Günter U. Höglinger |
| author_facet | Matthias Höllerhage Linghan Duan Oscar Wing Ho Chua Claudia Moebius Svenja H. Bothe Kristina Losse Rebecca Kotzur Kristina Lau Franziska Hopfner Franziska Richter Christian Wahl-Schott Marc Bickle Günter U. Höglinger |
| author_sort | Matthias Höllerhage |
| collection | DOAJ |
| description | Abstract Background Alpha-synuclein (αSyn) is a major player in the pathophysiology of synucleinopathies, which include Parkinson’s disease, dementia with Lewy bodies, and multiple system atrophy. To date, there is no disease-modifying therapy available for these synucleinopathies. Furthermore, the intracellular mechanisms by which αSyn confers toxicity are not yet fully understood. Therefore, it is of utmost importance to investigate the pathophysiology of αSyn-induced toxicity in order to identify novel molecular targets for the development of disease-modifying therapies. Methods We performed the first genome-wide siRNA modifier screening in a human postmitotic neuronal cell model using αSyn-induced toxicity as a read-out. In a multi-step approach, we identified several genes, whose knockdown protected against αSyn-induced toxicity. The main hit was further validated by different methods, including immunofluorescence microscopy, qPCR, and Western blot. Furthermore, the main finding was confirmed in mouse primary neurons. Results The highest protection was achieved by knockdown of SNX5, which encodes the sorting nexin 5 (SNX5) protein, a component of the retromer complex. The protective efficacy of SNX5 knockdown was confirmed with an independent siRNA system. The protective effect of SNX5 knockdown was further confirmed in primary neurons from transgenic mice, where the knockdown of SNX5 led to amelioration of decrease in synchrony that was observed in untreated and control-siRNA-treated cells. SNX5 protein is a component of the SNX-BAR (Bin/Amphiphysin/Rvs) heterodimer, which is part of the retromer complex. Extracellular αSyn and overexpression of intracellular αSyn led to fragmentation of the trans-Golgi network, which was prevented by SNX5 knockdown that led to confinement of αSyn in early endosomes. Conclusion In summary, our data suggest that SNX5 plays an important role in the trafficking and toxicity of αSyn. Therefore, SNX5 appears to be a target of therapeutic intervention for synucleinopathies. |
| format | Article |
| id | doaj-art-c09582e6d0a843e7bd5fca98294ae4ef |
| institution | Kabale University |
| issn | 2047-9158 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | BMC |
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| series | Translational Neurodegeneration |
| spelling | doaj-art-c09582e6d0a843e7bd5fca98294ae4ef2025-08-20T03:25:19ZengBMCTranslational Neurodegeneration2047-91582025-06-0114111910.1186/s40035-025-00486-5A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell modelMatthias Höllerhage0Linghan Duan1Oscar Wing Ho Chua2Claudia Moebius3Svenja H. Bothe4Kristina Losse5Rebecca Kotzur6Kristina Lau7Franziska Hopfner8Franziska Richter9Christian Wahl-Schott10Marc Bickle11Günter U. Höglinger12Department of Neurology, Hannover Medical SchoolDepartment of Neurology, Hannover Medical SchoolDepartment of Neurology, Hannover Medical SchoolHT-Technology Development Studio, Max Planck Institute of Molecular Cell Biology and GeneticsDepartment of Neurology, Hannover Medical SchoolDepartment of Neurology, Hannover Medical SchoolDepartment of Pharmacology, Toxicology and Pharmacy, University of Veterinary Medicine HannoverCenter for Systems NeuroscienceDepartment of Neurology, LMU University Hospital, Ludwig-Maximilians-Universität (LMU)Center for Systems NeuroscienceThe Institute of Cardiovascular Physiology and Pathophysiology, Ludwig-Maximilians-Universität (LMU)HT-Technology Development Studio, Max Planck Institute of Molecular Cell Biology and GeneticsDepartment of Neurology, LMU University Hospital, Ludwig-Maximilians-Universität (LMU)Abstract Background Alpha-synuclein (αSyn) is a major player in the pathophysiology of synucleinopathies, which include Parkinson’s disease, dementia with Lewy bodies, and multiple system atrophy. To date, there is no disease-modifying therapy available for these synucleinopathies. Furthermore, the intracellular mechanisms by which αSyn confers toxicity are not yet fully understood. Therefore, it is of utmost importance to investigate the pathophysiology of αSyn-induced toxicity in order to identify novel molecular targets for the development of disease-modifying therapies. Methods We performed the first genome-wide siRNA modifier screening in a human postmitotic neuronal cell model using αSyn-induced toxicity as a read-out. In a multi-step approach, we identified several genes, whose knockdown protected against αSyn-induced toxicity. The main hit was further validated by different methods, including immunofluorescence microscopy, qPCR, and Western blot. Furthermore, the main finding was confirmed in mouse primary neurons. Results The highest protection was achieved by knockdown of SNX5, which encodes the sorting nexin 5 (SNX5) protein, a component of the retromer complex. The protective efficacy of SNX5 knockdown was confirmed with an independent siRNA system. The protective effect of SNX5 knockdown was further confirmed in primary neurons from transgenic mice, where the knockdown of SNX5 led to amelioration of decrease in synchrony that was observed in untreated and control-siRNA-treated cells. SNX5 protein is a component of the SNX-BAR (Bin/Amphiphysin/Rvs) heterodimer, which is part of the retromer complex. Extracellular αSyn and overexpression of intracellular αSyn led to fragmentation of the trans-Golgi network, which was prevented by SNX5 knockdown that led to confinement of αSyn in early endosomes. Conclusion In summary, our data suggest that SNX5 plays an important role in the trafficking and toxicity of αSyn. Therefore, SNX5 appears to be a target of therapeutic intervention for synucleinopathies.https://doi.org/10.1186/s40035-025-00486-5Genome-wide RNAi screeningParkinson’s diseaseAlpha-synucleinRetromerSNX5Trans-Golgi network |
| spellingShingle | Matthias Höllerhage Linghan Duan Oscar Wing Ho Chua Claudia Moebius Svenja H. Bothe Kristina Losse Rebecca Kotzur Kristina Lau Franziska Hopfner Franziska Richter Christian Wahl-Schott Marc Bickle Günter U. Höglinger A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model Translational Neurodegeneration Genome-wide RNAi screening Parkinson’s disease Alpha-synuclein Retromer SNX5 Trans-Golgi network |
| title | A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model |
| title_full | A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model |
| title_fullStr | A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model |
| title_full_unstemmed | A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model |
| title_short | A genome-wide RNA interference screening reveals protectiveness of SNX5 knockdown in a Parkinson’s disease cell model |
| title_sort | genome wide rna interference screening reveals protectiveness of snx5 knockdown in a parkinson s disease cell model |
| topic | Genome-wide RNAi screening Parkinson’s disease Alpha-synuclein Retromer SNX5 Trans-Golgi network |
| url | https://doi.org/10.1186/s40035-025-00486-5 |
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