Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease
Background Aggregation of the amyloid-β (Aβ) peptide in the brain is one of the key pathological events in Alzheimer’s disease (AD). Reducing Aβ levels in the brain by enhancing its degradation is one possible strategy to develop new therapies for AD. Neprilysin (NEP) is a membrane-bound metallopept...
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Uppsala universitet, Institutionen för farmaci
2022
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Online Access: | http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-472469 https://doi.org/10.1186/s13195-022-01132-2 |
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ftuppsalauniv:oai:DiVA.org:uu-472469 2023-05-15T15:18:30+02:00 Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease Rofo, Fadi Metzendorf, Nicole Saubi, Cristina Suominen, Laura Godec, Ana Sehlin, Dag Syvänen, Stina Hultqvist, Greta 2022 application/pdf http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-472469 https://doi.org/10.1186/s13195-022-01132-2 eng eng Uppsala universitet, Institutionen för farmaci Uppsala universitet, Geriatrik Alzheimer's Research & Therapy, 2022, 14:1, orcid:0000-0003-2249-653x orcid:0000-0003-1163-0182 orcid:0000-0002-9430-3859 orcid:0000-0002-8196-4041 orcid:0000-0002-4136-6792 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-472469 doi:10.1186/s13195-022-01132-2 PMID 36471433 ISI:000894439900002 info:eu-repo/semantics/openAccess Amyloid-β (Aβ) neprilysin (NEP) blood-brain barrier (BBB) transferrin receptor (TfR) recombinant proteins Neurosciences Neurovetenskaper Pharmaceutical Sciences Farmaceutiska vetenskaper Article in journal info:eu-repo/semantics/article text 2022 ftuppsalauniv https://doi.org/10.1186/s13195-022-01132-2 2023-02-23T22:01:23Z Background Aggregation of the amyloid-β (Aβ) peptide in the brain is one of the key pathological events in Alzheimer’s disease (AD). Reducing Aβ levels in the brain by enhancing its degradation is one possible strategy to develop new therapies for AD. Neprilysin (NEP) is a membrane-bound metallopeptidase and one of the major Aβ-degrading enzymes. The secreted soluble form of NEP (sNEP) has been previously suggested as a potential protein-therapy degrading Aβ in AD. However, similar to other large molecules, peripherally administered sNEP is unable to reach the brain due to the presence of the blood–brain barrier (BBB). Methods To provide transcytosis across the BBB, we recombinantly fused the TfR binding moiety (scFv8D3) to either sNEP or a previously described variant of NEP (muNEP) suggested to have higher degradation efficiency of Aβ compared to other NEP substrates, but not per se to degrade Aβ more efficiently. To provide long blood half-life, an Fc-based antibody fragment (scFc) was added to the designs, forming sNEP-scFc-scFv8D3 and muNEP-scFc-scFv8D3. The ability of the mentioned recombinant proteins to degrade Aβ was first evaluated in vitro using synthetic Aβ peptides followed by sandwich ELISA. For the in vivo studies, a single injection of 125-iodine-labelled sNEP-scFc-scFv8D3 and muNEP-scFc-scFv8D3 was intravenously administered to a tg-ArcSwe mouse model of AD, using scFc-scFv8D3 protein that lacks NEP as a negative control. Different ELISA setups were applied to quantify Aβ concentration of different conformations, both in brain tissues and blood samples. Results When tested in vitro, sNEP-scFc-scFv8D3 retained sNEP enzymatic activity in degrading Aβ and both constructs efficiently degraded arctic Aβ. When intravenously injected, sNEP-scFc-scFv8D3 demonstrated 20 times higher brain uptake compared to sNEP. Both scFv8D3-fused NEP proteins significantly reduced aggregated Aβ levels in the blood of tg-ArcSwe mice, a transgenic mouse model of AD, following a single intravenous injection. In the brain, ... Article in Journal/Newspaper Arctic Uppsala University: Publications (DiVA) Arctic Alzheimer's Research & Therapy 14 1 |
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Uppsala University: Publications (DiVA) |
op_collection_id |
ftuppsalauniv |
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English |
topic |
Amyloid-β (Aβ) neprilysin (NEP) blood-brain barrier (BBB) transferrin receptor (TfR) recombinant proteins Neurosciences Neurovetenskaper Pharmaceutical Sciences Farmaceutiska vetenskaper |
spellingShingle |
Amyloid-β (Aβ) neprilysin (NEP) blood-brain barrier (BBB) transferrin receptor (TfR) recombinant proteins Neurosciences Neurovetenskaper Pharmaceutical Sciences Farmaceutiska vetenskaper Rofo, Fadi Metzendorf, Nicole Saubi, Cristina Suominen, Laura Godec, Ana Sehlin, Dag Syvänen, Stina Hultqvist, Greta Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease |
topic_facet |
Amyloid-β (Aβ) neprilysin (NEP) blood-brain barrier (BBB) transferrin receptor (TfR) recombinant proteins Neurosciences Neurovetenskaper Pharmaceutical Sciences Farmaceutiska vetenskaper |
description |
Background Aggregation of the amyloid-β (Aβ) peptide in the brain is one of the key pathological events in Alzheimer’s disease (AD). Reducing Aβ levels in the brain by enhancing its degradation is one possible strategy to develop new therapies for AD. Neprilysin (NEP) is a membrane-bound metallopeptidase and one of the major Aβ-degrading enzymes. The secreted soluble form of NEP (sNEP) has been previously suggested as a potential protein-therapy degrading Aβ in AD. However, similar to other large molecules, peripherally administered sNEP is unable to reach the brain due to the presence of the blood–brain barrier (BBB). Methods To provide transcytosis across the BBB, we recombinantly fused the TfR binding moiety (scFv8D3) to either sNEP or a previously described variant of NEP (muNEP) suggested to have higher degradation efficiency of Aβ compared to other NEP substrates, but not per se to degrade Aβ more efficiently. To provide long blood half-life, an Fc-based antibody fragment (scFc) was added to the designs, forming sNEP-scFc-scFv8D3 and muNEP-scFc-scFv8D3. The ability of the mentioned recombinant proteins to degrade Aβ was first evaluated in vitro using synthetic Aβ peptides followed by sandwich ELISA. For the in vivo studies, a single injection of 125-iodine-labelled sNEP-scFc-scFv8D3 and muNEP-scFc-scFv8D3 was intravenously administered to a tg-ArcSwe mouse model of AD, using scFc-scFv8D3 protein that lacks NEP as a negative control. Different ELISA setups were applied to quantify Aβ concentration of different conformations, both in brain tissues and blood samples. Results When tested in vitro, sNEP-scFc-scFv8D3 retained sNEP enzymatic activity in degrading Aβ and both constructs efficiently degraded arctic Aβ. When intravenously injected, sNEP-scFc-scFv8D3 demonstrated 20 times higher brain uptake compared to sNEP. Both scFv8D3-fused NEP proteins significantly reduced aggregated Aβ levels in the blood of tg-ArcSwe mice, a transgenic mouse model of AD, following a single intravenous injection. In the brain, ... |
format |
Article in Journal/Newspaper |
author |
Rofo, Fadi Metzendorf, Nicole Saubi, Cristina Suominen, Laura Godec, Ana Sehlin, Dag Syvänen, Stina Hultqvist, Greta |
author_facet |
Rofo, Fadi Metzendorf, Nicole Saubi, Cristina Suominen, Laura Godec, Ana Sehlin, Dag Syvänen, Stina Hultqvist, Greta |
author_sort |
Rofo, Fadi |
title |
Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease |
title_short |
Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease |
title_full |
Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease |
title_fullStr |
Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease |
title_full_unstemmed |
Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease |
title_sort |
blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of alzheimer’s disease |
publisher |
Uppsala universitet, Institutionen för farmaci |
publishDate |
2022 |
url |
http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-472469 https://doi.org/10.1186/s13195-022-01132-2 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_relation |
Alzheimer's Research & Therapy, 2022, 14:1, orcid:0000-0003-2249-653x orcid:0000-0003-1163-0182 orcid:0000-0002-9430-3859 orcid:0000-0002-8196-4041 orcid:0000-0002-4136-6792 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-472469 doi:10.1186/s13195-022-01132-2 PMID 36471433 ISI:000894439900002 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1186/s13195-022-01132-2 |
container_title |
Alzheimer's Research & Therapy |
container_volume |
14 |
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1 |
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1766348695882694656 |