Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent
Abstract Unlike variant Creutzfeldt–Jakob disease prions, sporadic Creutzfeldt–Jakob disease prions have been shown to be difficult to amplify in vitro by protein misfolding cyclic amplification (PMCA). We assessed PMCA of pathological prion protein (PrPTSE) from 14 human sCJD brain samples in 3 sub...
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2021-02-01
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doaj-49816cda18ac44b7b8c29880c70453922021-02-21T12:33:19ZengNature Publishing GroupScientific Reports2045-23222021-02-0111111210.1038/s41598-021-83630-1Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependentMaxime Bélondrade0Simon Nicot1Charly Mayran2Lilian Bruyere-Ostells3Florian Almela4Michele A. Di Bari5Etienne Levavasseur6Joel C. Watts7Chantal Fournier-Wirth8Sylvain Lehmann9Stéphane Haïk10Romolo Nonno11Daisy Bougard12Pathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierPathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierPathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierPathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierPathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierDepartment of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di SanitaInserm U 1127, CNRS UMR 7225, UPMC Université Paris 06 UMR S 1127, Institut du Cerveau et de la Moelle épinière, Sorbonne UniversitésTanz Centre for Research in Neurodegenerative Diseases and Department of Biochemistry, University of TorontoPathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierIRMB, INM, INSERM, CHU Montpellier, (LBPC-PPC), Univ MontpellierInserm U 1127, CNRS UMR 7225, UPMC Université Paris 06 UMR S 1127, Institut du Cerveau et de la Moelle épinière, Sorbonne UniversitésDepartment of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di SanitaPathogenesis and Control of Chronic Infections, Etablissement Français du Sang, Inserm, Université de MontpellierAbstract Unlike variant Creutzfeldt–Jakob disease prions, sporadic Creutzfeldt–Jakob disease prions have been shown to be difficult to amplify in vitro by protein misfolding cyclic amplification (PMCA). We assessed PMCA of pathological prion protein (PrPTSE) from 14 human sCJD brain samples in 3 substrates: 2 from transgenic mice expressing human prion protein (PrP) with either methionine (M) or valine (V) at position 129, and 1 from bank voles. Brain extracts representing the 5 major clinicopathological sCJD subtypes (MM1/MV1, MM2, MV2, VV1, and VV2) all triggered seeded PrPTSE amplification during serial PMCA with strong seed- and substrate-dependence. Remarkably, bank vole PrP substrate allowed the propagation of all sCJD subtypes with preservation of the initial molecular PrPTSE type. In contrast, PMCA in human PrP substrates was accompanied by a PrPTSE molecular shift during heterologous (M/V129) PMCA reactions, with increased permissiveness of V129 PrP substrate to in vitro sCJD prion amplification compared to M129 PrP substrate. Combining PMCA amplification sensitivities with PrPTSE electrophoretic profiles obtained in the different substrates confirmed the classification of 4 distinct major sCJD prion strains (M1, M2, V1, and V2). Finally, the level of sensitivity required to detect VV2 sCJD prions in cerebrospinal fluid was achieved.https://doi.org/10.1038/s41598-021-83630-1 |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Maxime Bélondrade Simon Nicot Charly Mayran Lilian Bruyere-Ostells Florian Almela Michele A. Di Bari Etienne Levavasseur Joel C. Watts Chantal Fournier-Wirth Sylvain Lehmann Stéphane Haïk Romolo Nonno Daisy Bougard |
spellingShingle |
Maxime Bélondrade Simon Nicot Charly Mayran Lilian Bruyere-Ostells Florian Almela Michele A. Di Bari Etienne Levavasseur Joel C. Watts Chantal Fournier-Wirth Sylvain Lehmann Stéphane Haïk Romolo Nonno Daisy Bougard Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent Scientific Reports |
author_facet |
Maxime Bélondrade Simon Nicot Charly Mayran Lilian Bruyere-Ostells Florian Almela Michele A. Di Bari Etienne Levavasseur Joel C. Watts Chantal Fournier-Wirth Sylvain Lehmann Stéphane Haïk Romolo Nonno Daisy Bougard |
author_sort |
Maxime Bélondrade |
title |
Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent |
title_short |
Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent |
title_full |
Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent |
title_fullStr |
Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent |
title_full_unstemmed |
Sensitive protein misfolding cyclic amplification of sporadic Creutzfeldt–Jakob disease prions is strongly seed and substrate dependent |
title_sort |
sensitive protein misfolding cyclic amplification of sporadic creutzfeldt–jakob disease prions is strongly seed and substrate dependent |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-02-01 |
description |
Abstract Unlike variant Creutzfeldt–Jakob disease prions, sporadic Creutzfeldt–Jakob disease prions have been shown to be difficult to amplify in vitro by protein misfolding cyclic amplification (PMCA). We assessed PMCA of pathological prion protein (PrPTSE) from 14 human sCJD brain samples in 3 substrates: 2 from transgenic mice expressing human prion protein (PrP) with either methionine (M) or valine (V) at position 129, and 1 from bank voles. Brain extracts representing the 5 major clinicopathological sCJD subtypes (MM1/MV1, MM2, MV2, VV1, and VV2) all triggered seeded PrPTSE amplification during serial PMCA with strong seed- and substrate-dependence. Remarkably, bank vole PrP substrate allowed the propagation of all sCJD subtypes with preservation of the initial molecular PrPTSE type. In contrast, PMCA in human PrP substrates was accompanied by a PrPTSE molecular shift during heterologous (M/V129) PMCA reactions, with increased permissiveness of V129 PrP substrate to in vitro sCJD prion amplification compared to M129 PrP substrate. Combining PMCA amplification sensitivities with PrPTSE electrophoretic profiles obtained in the different substrates confirmed the classification of 4 distinct major sCJD prion strains (M1, M2, V1, and V2). Finally, the level of sensitivity required to detect VV2 sCJD prions in cerebrospinal fluid was achieved. |
url |
https://doi.org/10.1038/s41598-021-83630-1 |
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