High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy
Mitochondrial dysfunction and aberrant mitochondrial homeostasis are key aspects of Parkinson's disease (PD) pathophysiology. Mutations in PINK1 and Parkin proteins lead to autosomal recessive PD, suggesting that defective mitochondrial clearance via mitophagy is key in PD etiology. Acceleratin...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-04-01
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| Series: | SLAS Discovery |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472555222137160 |
| _version_ | 1797842462740316160 |
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| author | Roberta Tufi Emily H. Clark Tamaki Hoshikawa Christiana Tsagkaraki Jack Stanley Kunitoshi Takeda James M. Staddon Thomas Briston |
| author_facet | Roberta Tufi Emily H. Clark Tamaki Hoshikawa Christiana Tsagkaraki Jack Stanley Kunitoshi Takeda James M. Staddon Thomas Briston |
| author_sort | Roberta Tufi |
| collection | DOAJ |
| description | Mitochondrial dysfunction and aberrant mitochondrial homeostasis are key aspects of Parkinson's disease (PD) pathophysiology. Mutations in PINK1 and Parkin proteins lead to autosomal recessive PD, suggesting that defective mitochondrial clearance via mitophagy is key in PD etiology. Accelerating the identification and/or removal of dysfunctional mitochondria could therefore provide a disease-modifying approach to treatment. To that end, we performed a high-content phenotypic screen (HCS) of ∼125,000 small molecules to identify compounds that positively modulate mitochondrial accumulation of the PINK1-Parkin-dependent mitophagy initiation marker p-Ser65-Ub in Parkin haploinsufficiency (Parkin +/R275W) human fibroblasts. Following confirmatory counter-screening and orthogonal assays, we selected compounds of interest that enhance mitophagy-related biochemical and functional endpoints in patient-derived fibroblasts. Identification of inhibitors of the ubiquitin-specific peptidase and negative regulator of mitophagy USP30 within our hits further validated our approach. The compounds identified in this work provide a novel starting point for further investigation and optimization. |
| first_indexed | 2024-04-09T16:48:18Z |
| format | Article |
| id | doaj.art-baab2dcd99a8404dbebb837a80359c4c |
| institution | Directory Open Access Journal |
| issn | 2472-5552 |
| language | English |
| last_indexed | 2024-04-09T16:48:18Z |
| publishDate | 2023-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj.art-baab2dcd99a8404dbebb837a80359c4c2023-04-22T06:22:04ZengElsevierSLAS Discovery2472-55522023-04-012837387High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagyRoberta Tufi0Emily H. Clark1Tamaki Hoshikawa2Christiana Tsagkaraki3Jack Stanley4Kunitoshi Takeda5James M. Staddon6Thomas Briston7Neurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomNeurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomNeurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomNeurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomNeurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomNeurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomNeurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomCorresponding author.; Neurology Innovation Centre, Hatfield Research Laboratories, Eisai Ltd., Hatfield AL10 9SN, United KingdomMitochondrial dysfunction and aberrant mitochondrial homeostasis are key aspects of Parkinson's disease (PD) pathophysiology. Mutations in PINK1 and Parkin proteins lead to autosomal recessive PD, suggesting that defective mitochondrial clearance via mitophagy is key in PD etiology. Accelerating the identification and/or removal of dysfunctional mitochondria could therefore provide a disease-modifying approach to treatment. To that end, we performed a high-content phenotypic screen (HCS) of ∼125,000 small molecules to identify compounds that positively modulate mitochondrial accumulation of the PINK1-Parkin-dependent mitophagy initiation marker p-Ser65-Ub in Parkin haploinsufficiency (Parkin +/R275W) human fibroblasts. Following confirmatory counter-screening and orthogonal assays, we selected compounds of interest that enhance mitophagy-related biochemical and functional endpoints in patient-derived fibroblasts. Identification of inhibitors of the ubiquitin-specific peptidase and negative regulator of mitophagy USP30 within our hits further validated our approach. The compounds identified in this work provide a novel starting point for further investigation and optimization.http://www.sciencedirect.com/science/article/pii/S2472555222137160High-content screeningMitophagyParkinParkinson's diseasePINK1USP30 |
| spellingShingle | Roberta Tufi Emily H. Clark Tamaki Hoshikawa Christiana Tsagkaraki Jack Stanley Kunitoshi Takeda James M. Staddon Thomas Briston High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy SLAS Discovery High-content screening Mitophagy Parkin Parkinson's disease PINK1 USP30 |
| title | High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy |
| title_full | High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy |
| title_fullStr | High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy |
| title_full_unstemmed | High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy |
| title_short | High-content phenotypic screen to identify small molecule enhancers of Parkin-dependent ubiquitination and mitophagy |
| title_sort | high content phenotypic screen to identify small molecule enhancers of parkin dependent ubiquitination and mitophagy |
| topic | High-content screening Mitophagy Parkin Parkinson's disease PINK1 USP30 |
| url | http://www.sciencedirect.com/science/article/pii/S2472555222137160 |
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