Identification and in vivo characterization of a brain-penetrating nanobody

Fluids and Barriers of the CNS

T Jaspers¹  Y Wouters¹  B De Strooper²  M Dewilde³ 

[1] VIB Center for Brain & Disease Research, Campus Gasthuisberg O&N4, Herestraat 49, box 602, B-3000, Leuven, Belgium;Laboratory for the Research of Neurodegenerative Diseases, Department of Neurosciences, Leuven Brain Institute (LBI), KU Leuven, B-3000, Leuven, Belgium;VIB Center for Brain & Disease Research, Campus Gasthuisberg O&N4, Herestraat 49, box 602, B-3000, Leuven, Belgium;Laboratory for the Research of Neurodegenerative Diseases, Department of Neurosciences, Leuven Brain Institute (LBI), KU Leuven, B-3000, Leuven, Belgium;UK Dementia Research Institute, University College London, London, UK;VIB Center for Brain & Disease Research, Campus Gasthuisberg O&N4, Herestraat 49, box 602, B-3000, Leuven, Belgium;Laboratory for the Research of Neurodegenerative Diseases, Department of Neurosciences, Leuven Brain Institute (LBI), KU Leuven, B-3000, Leuven, Belgium;VIB Discovery Sciences, B-3000, Leuven, Belgium;Laboratory for Therapeutic and Diagnostic Antibodies, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, B-3000, Leuven, Belgium;

关键词: Nanobody;    VHH;    Transferrin receptor;    Neurotensin;    Blood–brain barrier;    Receptor-mediated transcytosis;   

DOI  :  10.1186/s12987-020-00226-z

来源: Springer

【 摘 要 】

BackgroundPreclinical 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.MethodsVarious 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.ResultsOne 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.ConclusionThis 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.

【 授权许可】

CC BY