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Identification and in vivo characterization of a brain-penetrating nanobody
Identification and in vivo characterization of a brain-penetrating nanobody
- Source :
- Fluids and Barriers of the CNS, Vol 17, Iss 1, Pp 1-10 (2020), Fluids and Barriers of the CNS
- Publication Year :
- 2020
- Publisher :
- BMC, 2020.
-
Abstract
- Background Preclinical models to determine blood to brain transport ability of therapeutics are often ambiguous. In this study a method is developed that relies on CNS target-engagement and is able to rank brain-penetrating capacities. This method led to the discovery of an anti-transferrin receptor nanobody that is able to deliver a biologically active peptide to the brain via receptor-mediated transcytosis. Methods Various nanobodies against the mouse transferrin receptor were fused to neurotensin and injected peripherally in mice. Neurotensin is a neuropeptide that causes hypothermia when present in the brain but is unable to reach the brain from the periphery. Continuous body temperature measurements were used as a readout for brain penetration of nanobody-neurotensin fusions after its peripheral administration. Full temperature curves were analyzed using two-way ANOVA with Dunnett multiple comparisons tests. Results One anti-transferrin receptor nanobody coupled to neurotensin elicited a drop in body temperature following intravenous injection. Epitope binning indicated that this nanobody bound a distinct transferrin receptor epitope compared to the non-crossing nanobodies. This brain-penetrating nanobody was used to characterize the in vivo hypothermia model. The hypothermic effect caused by neurotensin is dose-dependent and could be used to directly compare peripheral administration routes and various nanobodies in terms of brain exposure. Conclusion This method led to the discovery of an anti-transferrin receptor nanobody that can reach the brain via receptor-mediated transcytosis after peripheral administration. This method could be used to assess novel proteins for brain-penetrating capabilities using a target-engaging readout.
- Subjects :
- Male
0301 basic medicine
Transferrin receptor
Neuropeptide
VHH
Blood–brain barrier
Receptor-mediated transcytosis
lcsh:RC346-429
Body Temperature
Mice
03 medical and health sciences
Cellular and Molecular Neuroscience
chemistry.chemical_compound
0302 clinical medicine
Developmental Neuroscience
In vivo
Receptors, Transferrin
Epitope binning
medicine
Animals
Receptors, Neurotensin
Receptor
Neurotensin
lcsh:Neurology. Diseases of the nervous system
Chemistry
Research
Brain
General Medicine
Single-Domain Antibodies
Cell biology
030104 developmental biology
medicine.anatomical_structure
Neurology
Transcytosis
Nanobody
Female
Camelids, New World
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 20458118
- Volume :
- 17
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Fluids and Barriers of the CNS
- Accession number :
- edsair.doi.dedup.....d3aaf48a57ed92d3e0c297cfea31e152
- Full Text :
- https://doi.org/10.1186/s12987-020-00226-z