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However, how lysosomes participate in the spreading of \u03b1-syn aggregates is unclear. Here, by using super-resolution (SR) and electron microscopy (EM), we find that \u03b1-syn fibrils affect the morphology of lysosomes and impair their function in neuronal cells. In addition, we demonstrate that \u03b1-syn fibrils induce peripheral redistribution of lysosomes, likely mediated by transcription factor EB (TFEB), increasing the efficiency of \u03b1-syn fibrils\u2019 transfer to neighboring cells. We also show that lysosomal membrane permeabilization (LMP) allows the seeding of soluble \u03b1-syn in cells that have taken up \u03b1-syn fibrils from the culture medium, and, more importantly, in healthy cells in coculture, following lysosome-mediated transfer of the fibrils. Moreover, we demonstrate that seeding occurs mainly at lysosomes in both donor and acceptor cells, after uptake of \u03b1-syn fibrils from the medium and following their transfer, respectively. Finally, by using a heterotypic coculture system, we determine the origin and nature of the lysosomes transferred between cells, and we show that donor cells bearing \u03b1-syn fibrils transfer damaged lysosomes to acceptor cells, while also receiving healthy lysosomes from them. These findings thus contribute to the elucidation of the mechanism by which \u03b1-syn fibrils spread through TNTs, while also revealing the crucial role of lysosomes, working as a Trojan horse for both seeding and propagation of disease pathology.<\/jats:p>","DOI":"10.1371\/journal.pbio.3001287","type":"journal-article","created":{"date-parts":[[2021,7,20]],"date-time":"2021-07-20T13:54:14Z","timestamp":1626789254000},"page":"e3001287","update-policy":"https:\/\/doi.org\/10.1371\/journal.pbio.corrections_policy","source":"Crossref","is-referenced-by-count":131,"title":["\u03b1-Synuclein fibrils subvert lysosome structure and function for the propagation of protein misfolding between cells through tunneling nanotubes"],"prefix":"10.1371","volume":"19","author":[{"given":"Aysegul","family":"Dilsizoglu Senol","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6376-5524","authenticated-orcid":true,"given":"Maura","family":"Samarani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvie","family":"Syan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4662-9360","authenticated-orcid":true,"given":"Carlos M.","family":"Guardia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0830-9403","authenticated-orcid":true,"given":"Takashi","family":"Nonaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nalan","family":"Liv","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Latour-Lambert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masato","family":"Hasegawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4835-6228","authenticated-orcid":true,"given":"Judith","family":"Klumperman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan S.","family":"Bonifacino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6048-6602","authenticated-orcid":true,"given":"Chiara","family":"Zurzolo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,7,20]]},"reference":[{"key":"pbio.3001287.ref001","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.1038\/s41593-018-0235-9","article-title":"Protein misfolding, aggregation, and conformational strains in neurodegenerative diseases","volume":"21","author":"C Soto","year":"2018","journal-title":"Nat Neurosci"},{"key":"pbio.3001287.ref002","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.1038\/s41593-018-0237-7","article-title":"Converging pathways in neurodegeneration, from genetics to mechanisms","volume":"21","author":"L Gan","year":"2018","journal-title":"Nat Neurosci"},{"key":"pbio.3001287.ref003","first-page":"52","article-title":"Nature Reviews Disease Primers","author":"W. 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