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  High-throughput imaging of ATG9A distribution as a diagnostic functional assay for adaptor protein complex 4-associated hereditary spastic paraplegia

Ebrahimi-Fakhari, D., Alecu, J. E., Brechmann, B., Ziegler, M., Eberhardt, K., Jumo, H., et al. (2021). High-throughput imaging of ATG9A distribution as a diagnostic functional assay for adaptor protein complex 4-associated hereditary spastic paraplegia. Brain Communications, 3(4): fcab221. doi:10.1093/braincomms/fcab221.

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 Creators:
Ebrahimi-Fakhari, Darius1, Author
Alecu, Julian E.1, Author
Brechmann, Barbara1, Author
Ziegler, Marvin1, Author
Eberhardt, Kathrin1, Author
Jumo, Hellen1, Author
D'Amore, Angelica1, Author
Habibzadeh, Parham1, Author
Faghihi, Mohammad Ali1, Author
De Bleecker, Jan L.1, Author
Vuillaumier-Barrot, Sandrine1, Author
Auvin, Stephane1, Author
Santorelli, Filippo M.1, Author
Neuser, Sonja1, Author
Popp, Bernt1, Author
Yang, Edward1, Author
Barrett, Lee1, Author
Davies, Alexandra K.2, Author              
Saffari, Afshin1, Author
Hirst, Jennifer1, Author
Sahin, Mustafa1, Author more..
Affiliations:
1external, ou_persistent22              
2Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              

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Free keywords: NEURONAL AXONS; AUTOPHAGY; VARIANTS; AP-4; TRAFFICKING; MITOPHAGYNeurosciences & Neurology; hereditary spastic paraplegia; adaptor protein complex 4; functional assay; high-throughput imaging; biomarker;
 Abstract: Adaptor protein complex 4-associated hereditary spastic paraplegia is caused by biallelic loss-of-function variants in AP4B1, AP4M1, AP4E1 or AP4S1, which constitute the four subunits of this obligate complex. While the diagnosis of adaptor protein complex 4-associated hereditary spastic paraplegia relies on molecular testing, the interpretation of novel missense variants remains challenging. Here, we address this diagnostic gap by using patient-derived fibroblasts to establish a functional assay that measures the subcellular localization of ATG9A, a transmembrane protein that is sorted by adaptor protein complex 4. Using automated high-throughput microscopy, we determine the ratio of the ATG9A fluorescence in the trans-Golgi-network versus cytoplasm and ascertain that this metric meets standards for screening assays (Z'-factor robust >0.3, strictly standardized mean difference >3). The `ATG9A ratio' is increased in fibroblasts of 18 well-characterized adaptor protein complex 4-associated hereditary spastic paraplegia patients [mean: 1.54 +/- 0.13 versus 1.21 +/- 0.05 (standard deviation) in controls] and receiver-operating characteristic analysis demonstrates robust diagnostic power (area under the curve: 0.85, 95% confidence interval: 0.849-0.852). Using fibroblasts from two individuals with atypical clinical features and novel biallelic missense variants of unknown significance in AP4B1, we show that our assay can reliably detect adaptor protein complex 4 function. Our findings establish the 'ATG9A ratio' as a diagnostic marker of adaptor protein complex 4-associated hereditary spastic paraplegia.

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Language(s): eng - English
 Dates: 2021-09
 Publication Status: Published online
 Pages: 17
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Project name : Marie Sklodowska-Curie grant agreement
Grant ID : 896725
Funding program : Horizon 2020 (H2020)
Funding organization : European Commission (EC)

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Title: Brain Communications
Source Genre: Journal
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Publ. Info: Oxford : Oxford University Press
Pages: - Volume / Issue: 3 (4) Sequence Number: fcab221 Start / End Page: - Identifier: ISSN: 2632-1297
CoNE: https://pure.mpg.de/cone/journals/resource/2632-1297