VHHs as tools for therapeutic protein delivery to the central nervous system
Abstract Background The blood brain barrier (BBB) limits the therapeutic perspective for central nervous system (CNS) disorders. Previously we found an anti-mouse transferrin receptor (TfR) VHH (Nb62) that was able to deliver a biologically active neuropeptide into the CNS in mice. Here, we aimed to...
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BMC
2022-10-01
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Series: | Fluids and Barriers of the CNS |
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Online Access: | https://doi.org/10.1186/s12987-022-00374-4 |
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author | Yessica Wouters Tom Jaspers Laura Rué Lutgarde Serneels Bart De Strooper Maarten Dewilde |
author_facet | Yessica Wouters Tom Jaspers Laura Rué Lutgarde Serneels Bart De Strooper Maarten Dewilde |
author_sort | Yessica Wouters |
collection | DOAJ |
description | Abstract Background The blood brain barrier (BBB) limits the therapeutic perspective for central nervous system (CNS) disorders. Previously we found an anti-mouse transferrin receptor (TfR) VHH (Nb62) that was able to deliver a biologically active neuropeptide into the CNS in mice. Here, we aimed to test its potential to shuttle a therapeutic relevant cargo. Since this VHH could not recognize the human TfR and hence its translational potential is limited, we also aimed to find and validate an anti-human transferrin VHH to deliver a therapeutic cargo into the CNS. Methods Alpaca immunizations with human TfR, and subsequent phage selection and screening for human TfR binding VHHs was performed to find a human TfR specific VHH (Nb188). Its ability to cross the BBB was determined by fusing it to neurotensin, a neuropeptide that reduces body temperature when present in the CNS but is not able to cross the BBB on its own. Next, the anti–β-secretase 1 (BACE1) 1A11 Fab and Nb62 or Nb188 were fused to an Fc domain to generate heterodimeric antibodies (1A11AM-Nb62 and 1A11AM-Nb188). These were then administered intravenously in wild-type mice and in mice in which the murine apical domain of the TfR was replaced by the human apical domain (hAPI KI). Pharmacokinetic and pharmacodynamic (PK/PD) studies were performed to assess the concentration of the heterodimeric antibodies in the brain over time and the ability to inhibit brain-specific BACE1 by analysing the brain levels of Aβ1–40. Results Selections and screening of a phage library resulted in the discovery of an anti-human TfR VHH (Nb188). Fusion of Nb188 to neurotensin induced hypothermia after intravenous injections in hAPI KI mice. In addition, systemic administration 1A11AM-Nb62 and 1A11AM-Nb188 fusions were able to reduce Aβ1-40 levels in the brain whereas 1A11AM fused to an irrelevant VHH did not. A PK/PD experiment showed that this effect could last for 3 days. Conclusion We have discovered an anti-human TfR specific VHH that is able to reach the CNS when administered systemically. In addition, both the currently discovered anti-human TfR VHH and the previously identified mouse-specific anti-TfR VHH, are both able to shuttle a therapeutically relevant cargo into the CNS. We suggest the mouse-specific VHH as a valuable research tool in mice and the human-specific VHH as a moiety to enhance the delivery efficiency of therapeutics into the CNS in human patients. |
first_indexed | 2024-04-13T22:45:43Z |
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id | doaj.art-9416f90f5a1549e39005a8a4bb0b1d6c |
institution | Directory Open Access Journal |
issn | 2045-8118 |
language | English |
last_indexed | 2024-04-13T22:45:43Z |
publishDate | 2022-10-01 |
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series | Fluids and Barriers of the CNS |
spelling | doaj.art-9416f90f5a1549e39005a8a4bb0b1d6c2022-12-22T02:26:24ZengBMCFluids and Barriers of the CNS2045-81182022-10-0119111310.1186/s12987-022-00374-4VHHs as tools for therapeutic protein delivery to the central nervous systemYessica Wouters0Tom Jaspers1Laura Rué2Lutgarde Serneels3Bart De Strooper4Maarten Dewilde5VIB Center for Brain and Disease ResearchVIB Center for Brain and Disease ResearchVIB Center for Brain and Disease ResearchVIB Center for Brain and Disease ResearchVIB Center for Brain and Disease ResearchVIB Center for Brain and Disease ResearchAbstract Background The blood brain barrier (BBB) limits the therapeutic perspective for central nervous system (CNS) disorders. Previously we found an anti-mouse transferrin receptor (TfR) VHH (Nb62) that was able to deliver a biologically active neuropeptide into the CNS in mice. Here, we aimed to test its potential to shuttle a therapeutic relevant cargo. Since this VHH could not recognize the human TfR and hence its translational potential is limited, we also aimed to find and validate an anti-human transferrin VHH to deliver a therapeutic cargo into the CNS. Methods Alpaca immunizations with human TfR, and subsequent phage selection and screening for human TfR binding VHHs was performed to find a human TfR specific VHH (Nb188). Its ability to cross the BBB was determined by fusing it to neurotensin, a neuropeptide that reduces body temperature when present in the CNS but is not able to cross the BBB on its own. Next, the anti–β-secretase 1 (BACE1) 1A11 Fab and Nb62 or Nb188 were fused to an Fc domain to generate heterodimeric antibodies (1A11AM-Nb62 and 1A11AM-Nb188). These were then administered intravenously in wild-type mice and in mice in which the murine apical domain of the TfR was replaced by the human apical domain (hAPI KI). Pharmacokinetic and pharmacodynamic (PK/PD) studies were performed to assess the concentration of the heterodimeric antibodies in the brain over time and the ability to inhibit brain-specific BACE1 by analysing the brain levels of Aβ1–40. Results Selections and screening of a phage library resulted in the discovery of an anti-human TfR VHH (Nb188). Fusion of Nb188 to neurotensin induced hypothermia after intravenous injections in hAPI KI mice. In addition, systemic administration 1A11AM-Nb62 and 1A11AM-Nb188 fusions were able to reduce Aβ1-40 levels in the brain whereas 1A11AM fused to an irrelevant VHH did not. A PK/PD experiment showed that this effect could last for 3 days. Conclusion We have discovered an anti-human TfR specific VHH that is able to reach the CNS when administered systemically. In addition, both the currently discovered anti-human TfR VHH and the previously identified mouse-specific anti-TfR VHH, are both able to shuttle a therapeutically relevant cargo into the CNS. We suggest the mouse-specific VHH as a valuable research tool in mice and the human-specific VHH as a moiety to enhance the delivery efficiency of therapeutics into the CNS in human patients.https://doi.org/10.1186/s12987-022-00374-4NanobodyVHHTransferrin receptorNeurotensinBlood–brain barrierReceptor-mediated transcytosis |
spellingShingle | Yessica Wouters Tom Jaspers Laura Rué Lutgarde Serneels Bart De Strooper Maarten Dewilde VHHs as tools for therapeutic protein delivery to the central nervous system Fluids and Barriers of the CNS Nanobody VHH Transferrin receptor Neurotensin Blood–brain barrier Receptor-mediated transcytosis |
title | VHHs as tools for therapeutic protein delivery to the central nervous system |
title_full | VHHs as tools for therapeutic protein delivery to the central nervous system |
title_fullStr | VHHs as tools for therapeutic protein delivery to the central nervous system |
title_full_unstemmed | VHHs as tools for therapeutic protein delivery to the central nervous system |
title_short | VHHs as tools for therapeutic protein delivery to the central nervous system |
title_sort | vhhs as tools for therapeutic protein delivery to the central nervous system |
topic | Nanobody VHH Transferrin receptor Neurotensin Blood–brain barrier Receptor-mediated transcytosis |
url | https://doi.org/10.1186/s12987-022-00374-4 |
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