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Amyloid precursor protein products concentrate in a subset of exosomes specifically endocytosed by neurons

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Abstract

Amyloid beta peptide (Aβ), the main component of senile plaques of Alzheimer’s disease brains, is produced by sequential cleavage of amyloid precursor protein (APP) and of its C-terminal fragments (CTFs). An unanswered question is how amyloidogenic peptides spread throughout the brain during the course of the disease. Here, we show that small lipid vesicles called exosomes, secreted in the extracellular milieu by cortical neurons, carry endogenous APP and are strikingly enriched in CTF-α and the newly characterized CTF-η. Exosomes from N2a cells expressing human APP with the autosomal dominant Swedish mutation contain Aβ peptides as well as CTF-α and CTF-η, while those from cells expressing the non-mutated form of APP only contain CTF-α and CTF-η. APP and CTFs are sorted into a subset of exosomes which lack the tetraspanin CD63 and specifically bind to dendrites of neurons, unlike exosomes carrying CD63 which bind to both neurons and glial cells. Thus, neuroblastoma cells secrete distinct populations of exosomes carrying different cargoes and targeting specific cell types. APP-carrying exosomes can be endocytosed by receiving cells, allowing the processing of APP acquired by exosomes to give rise to the APP intracellular domain (AICD). Thus, our results show for the first time that neuronal exosomes may indeed act as vehicles for the intercellular transport of APP and its catabolites.

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Acknowledgements

We thank M. Jacquier-Sarlin, S. Fraboulet, M. Laporte, and K. Sadoul for critically reading the manuscript. This work was funded by Grants from the Agence Nationale de la Recherche (ANR MALZ program) and from Association France Alzheimer. K. Laulagnier was supported by the Fondation Plan Alzheimer. C. Javalet was supported by the Ministère de l’Enseignement Supérieur et de la Recherche.

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  1. Institut National de la Santé et de la Recherche Médicale (INSERM), U1216, 38042, Grenoble, France

    Karine Laulagnier, Charlotte Javalet, Fiona J. Hemming, Gaëlle Lachenal, Béatrice Blot, Christine Chatellard & Rémy Sadoul

  2. Institut des Neurosciences, Université Grenoble Alpes, 38042, Grenoble, France

    Karine Laulagnier, Charlotte Javalet, Fiona J. Hemming, Gaëlle Lachenal, Béatrice Blot, Christine Chatellard & Rémy Sadoul

  3. Dulbecco Telethon Institute Lab of Neurodegenerative Diseases, Centre for Integrative Biology (CIBIO), University of Trento, 38123, Trento, Italy

    Mathilde Chivet

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18_2017_2664_MOESM1_ESM.tif

Supplementary Fig. 1 Nanoparticle Tracking Analysis shows that APPwt or APPswe expression affect neither the number nor the size of secreted exosomes. Exosomes purified from supernatants of control N2a, APPwt N2a, and APPswe N2a were resuspended and analyzed using Nanosight. a Number of particles secreted during 24 h by 9 x 106 cells per dish. Averages of five measurements +/- SEM are shown. b Distribution of sizes for each type of exosomes. Average of five measurements is shown with SEM drawn in gray. The mode size is indicated for each population. No significant differences were observed between each type of exosomes. (TIFF 765 kb)

18_2017_2664_MOESM2_ESM.tif

Supplementary Fig. 2 N2a cells were transfected with UAS-Firefly luciferase reporter construct alone (Mock) or together with NLS-Gal4 construct (NLS-Gal4). Renilla luciferase was co-expressed to normalize Firefly activity and luciferase activities measured 48 h after transfection. When indicated, 10 μM of the γ-secretase inhibitor (DAPT) was added 20 h before measurement. In contrast with experiments using APP-Gal4 in Fig. 7, DAPT does not inhibit Firefly luciferase activity. The result represents the mean of two independent cultures. (TIFF 159 kb)

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Laulagnier, K., Javalet, C., Hemming, F.J. et al. Amyloid precursor protein products concentrate in a subset of exosomes specifically endocytosed by neurons. Cell. Mol. Life Sci. 75, 757–773 (2018). https://doi.org/10.1007/s00018-017-2664-0

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