Journal
FLUIDS AND BARRIERS OF THE CNS
Volume 17, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s12987-020-00226-z
Keywords
Nanobody; VHH; Transferrin receptor; Neurotensin; Blood-brain barrier; Receptor-mediated transcytosis
Categories
Funding
- Cure Alzheimer's Fund
- FWO [S007918N]
- KU Leuven
- VIB
- Flemish Government [3M140280]
- Geneeskundige Stichting Koningin Elisabeth
- MRC
- Alzheimer Society
- Alzheimer Research UK
- MRC [UKDRI-1004] Funding Source: UKRI
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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.
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