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[P2–054]: BRAIN DELIVERY OF AN AMYLOID‐β OLIGOMER (AβO)‐TARGETING PEPTIDE‐THERAPEUTIC BY A NOVEL BLOOD‐BRAIN BARRIER (BBB)‐CROSSING DOMAIN ANTIBODY
Author(s) -
Chakravarthy Balu,
Atkinson Trevor,
Brown Leslie,
Menard Michel,
Brunette Eric,
Pelletier Alex,
Jiang Susan,
Baumann Ewa,
Ball Marguerite,
Delaney Christie,
Ding Wen,
Star Alexandra,
RosaNeto Pedro,
Waterston Michael,
Yoganathan Nathan,
Parat Marie,
Grazzini Eric,
Haqqani Arsalan,
Lessard Etienne,
Whitfield James,
Rennie Kerry,
Durocher Yves,
Perkins Martin,
Stanimirovic Danica
Publication year - 2017
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2017.06.703
Subject(s) - blood–brain barrier , in vivo , western blot , in vitro , genetically modified mouse , fusion protein , chemistry , transgene , peptide , microbiology and biotechnology , recombinant dna , pharmacology , biology , biochemistry , endocrinology , central nervous system , gene
We have identified an amyloid-\udf (A\udf) binding peptide (ABP) that selectively bound pathological A\udf1-42 oligomers and reduced A\udf- mediated cell toxicity1. ABP also bound A\udf deposits in brains of AD transgenic mice and Alzheimer\u2019s disease (AD) patients in vitro, and when directly injected into live transgenic mice (APPSwe and \u394PS1) brain2, suggesting that, when delivered to the brain, ABP can engage A\udf and facilitate its clearance. Present study shows brain-delivery of ABP by a novel BBB-crossing domain antibody, and targeting and clearance of A\udf in transgenic mice. Recombinant ABP-BBB carrier fusion construct (ABP-BBB) were produced in CHO cells. BBB-permeability of the construct was assessed using in vitro BBB (formed by rat or human brain endothelial cells) and in vivo (rat and mouse) models. A\udf binding was determined by ELISA and Western blot overlay assays. For PK/PD studies, serum, CSF and brain levels of ABP-BBB construct and A\udf were assessed following iv injection by nanoLC- MRM, ELISA and Western blot methods. ABP-BBB bi-functional fusion protein was successfully expressed in CHO cells. The fusion protein retained both A\udf-oligomer binding activity and BBB-permeability in vitro. ABP-BBB transmigrated the in vitro BBB, in contrast to ABP without BBB carrier. When injected iv into rats, ABP-BBB appeared in the CSF in a dose- and time-dependent manner indicating transport of ABP across BBB in vivo. Apparent CNS exposure (Expapp), calculated as CSF[AUC]/serum[AUC], indicated a 15-20-fold increase in Expapp for ABP-BBB compared to non-BBB permeable ABP. This was further confirmed in mouse model wherein following iv injection, the fusion protein was detected in the CSF, and cortical and hippocampal regions of wild type and Tg mice. More importantly, in Tg mice ABP treatment resulted in a significant (\u223c50%) reduction of A\udf levels in the CSF, and brain cortex and hippocampus within 4-weeks.Peer reviewed: YesNRC publication: Ye