Blood–brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease
Abstract 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 me...
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ftdoajarticles:oai:doaj.org/article:a3ccdd56160f4deebd423b5623878295 2023-05-15T15:17:12+02:00 Blood–brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease Fadi Rofo Nicole G. Metzendorf Cristina Saubi Laura Suominen Ana Godec Dag Sehlin Stina Syvänen Greta Hultqvist 2022-12-01T00:00:00Z https://doi.org/10.1186/s13195-022-01132-2 https://doaj.org/article/a3ccdd56160f4deebd423b5623878295 EN eng BMC https://doi.org/10.1186/s13195-022-01132-2 https://doaj.org/toc/1758-9193 doi:10.1186/s13195-022-01132-2 1758-9193 https://doaj.org/article/a3ccdd56160f4deebd423b5623878295 Alzheimer’s Research & Therapy, Vol 14, Iss 1, Pp 1-19 (2022) Amyloid-β (Aβ) Neprilysin (NEP) Blood–brain barrier (BBB) Transferrin receptor (TfR) Recombinant proteins Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Neurology. Diseases of the nervous system RC346-429 article 2022 ftdoajarticles https://doi.org/10.1186/s13195-022-01132-2 2022-12-30T22:32:22Z Abstract 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 ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Alzheimer's Research & Therapy 14 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Amyloid-β (Aβ) Neprilysin (NEP) Blood–brain barrier (BBB) Transferrin receptor (TfR) Recombinant proteins Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Neurology. Diseases of the nervous system RC346-429 |
spellingShingle |
Amyloid-β (Aβ) Neprilysin (NEP) Blood–brain barrier (BBB) Transferrin receptor (TfR) Recombinant proteins Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Neurology. Diseases of the nervous system RC346-429 Fadi Rofo Nicole G. Metzendorf Cristina Saubi Laura Suominen Ana Godec Dag Sehlin Stina Syvänen Greta Hultqvist 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. Biological psychiatry. Neuropsychiatry RC321-571 Neurology. Diseases of the nervous system RC346-429 |
description |
Abstract 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 ... |
format |
Article in Journal/Newspaper |
author |
Fadi Rofo Nicole G. Metzendorf Cristina Saubi Laura Suominen Ana Godec Dag Sehlin Stina Syvänen Greta Hultqvist |
author_facet |
Fadi Rofo Nicole G. Metzendorf Cristina Saubi Laura Suominen Ana Godec Dag Sehlin Stina Syvänen Greta Hultqvist |
author_sort |
Fadi Rofo |
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 |
BMC |
publishDate |
2022 |
url |
https://doi.org/10.1186/s13195-022-01132-2 https://doaj.org/article/a3ccdd56160f4deebd423b5623878295 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Alzheimer’s Research & Therapy, Vol 14, Iss 1, Pp 1-19 (2022) |
op_relation |
https://doi.org/10.1186/s13195-022-01132-2 https://doaj.org/toc/1758-9193 doi:10.1186/s13195-022-01132-2 1758-9193 https://doaj.org/article/a3ccdd56160f4deebd423b5623878295 |
op_doi |
https://doi.org/10.1186/s13195-022-01132-2 |
container_title |
Alzheimer's Research & Therapy |
container_volume |
14 |
container_issue |
1 |
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1766347462663995392 |