Therapeutic bispecific antibodies cross the blood-brain barrier in nonhuman primates

Yu, Yang; Atwal, Jasvinder K.; Yin, Zhimin; Tong, Raymond K.; Wildsmith, Kristin R.; Tan, Christine; Bien-Ly, Nga; Hersom, Maria; Maloney, Janice; Meilandt, William J.; Bumbaca, Daniela; Gadkar, Kapil; Hoyte, Kwame; Luk, Wilman; Lu, Yanmei; Ernst, James A.; Scearce‐Levie, Kimberly; Couch, Jessica A.; Dennis, Mark S.; Watts, Ryan J.

doi:10.1126/scitranslmed.3009835

Cited by 296 publications

(305 citation statements)

References 16 publications

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“…A potential to deliver antibody therapeutics across the BBB, demonstrated recently with several BBB carrier antibodies, including TfR antibodies [5][6][7] and FC5, 11,13 has generated a renewed debate on mechanisms of internalization, intracellular traffic and polarized abluminal release of antibodies developed to cross the BBB via RMT. This study tracked the endocytic sorting and transcytosis of various internalizing and BBB-crossing antibody formats in BEC using a novel, quantitative mass spectrometry approach.…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, A20.1Fc showed slightly higher P app values in vitro and measurable CSF levels in vivo compared to A20.1, suggesting that Fc fragment may contribute to low, but detectable non-specific penetration of antibodies into the brain described in several studies. 5,6,13 A recent study 42 using high-resolution quantitative microscopy showed that lysosomal degradation of low levels of IgGs internalized into BEC from systemic circulations may be further limiting the antibody access to the brain.…”

Section: Discussionmentioning

confidence: 99%

“…Antibody or peptide ligands developed against these receptors have been exploited to 'transport' linked therapeutic 'cargo' including macromolecules across the BBB. Several antibodies have been developed as molecular Trojan horses for brain delivery of macromolecules, including various antibody formats against TfR, [4][5][6][7] humanized IgG against human insulin receptor (Insr), 8,9 antibodies against the heavy subunit of the large neutral aminoacid transporter (Lat1; Cd98) 10 and species cross-reactive camelid single-domain antibody, FC5. [11][12][13] Notably, FC5 has been shown to bind a glycosylated epitope of an antigen, putatively identified as Tmem30a, enriched in BEC, and to transmigrate across BEC monolayer via a saturable, directional, energy-and microtubule-dependent pathway, triggered by the antibody internalization via clathrin-coated vesicles.…”

Section: Introductionmentioning

confidence: 99%

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Endosomal trafficking regulates receptor-mediated transcytosis of antibodies across the blood brain barrier

Haqqani

Delaney

Brunette

et al. 2017

J Cereb Blood Flow Metab

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Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=33302487-9987-42d0-bb7f-3b59445d5f72 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=33302487-9987-42d0-bb7f-3b59445d5f72 AbstractCurrent methods for examining antibody trafficking are either non-quantitative such as immunocytochemistry or require antibody labeling with tracers. We have developed a multiplexed quantitative method for antibody 'tracking' in endosomal compartments of brain endothelial cells. Rat brain endothelial cells were co-incubated with blood-brain barrier (BBB)-crossing FC5, monovalent FC5Fc or bivalent FC5Fc fusion antibodies and control antibodies. Endosomes were separated using sucrose-density gradient ultracentrifugation and analyzed using multiplexed mass spectrometry to simultaneously quantify endosomal markers, receptor-mediated transcytosis (RMT) receptors and the co-incubated antibodies in each fraction. The quantitation showed that markers of early endosomes were enriched in high-density fractions (HDF), whereas markers of late endosomes and lysosomes were enriched in low-density fractions (LDF). RMT receptors, including transferrin receptor, showed a profile similar to that of early endosome markers. The in vitro BBB transcytosis rates of antibodies were directly proportional to their partition into early endosome fractions of brain endothelial cells. Addition of the Fc domain resulted in facilitated antibody 'redistribution' from LDF into HDF and additionally into multivesicular bodies (MVB). Sorting of various FC5 antibody formats away from late endosomes and lysosomes and into early endosomes and a subset of MVB results in increased antibody transcytosis at the abluminal side of the BBB.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Endosomal trafficking regulates receptor-mediated transcytosis of antibodies across the blood brain barrier

Haqqani

Delaney

Brunette

et al. 2017

J Cereb Blood Flow Metab

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“…Alternative scaffold-proteins are in general much smaller than full-length antibodies and can usually be engineered into multivalent as well as multispecific units with an overall size that remain smaller than the conventional antibody [17,18]. Such features can potentially improve in vivo biodistribution of the agent [19][20][21][22]. One type alternative binding-protein that has been investigated for various diagnostic and therapeutic applications is the small three-helical bundle Affibody molecule (6.5 kDa) [17,23].…”

mentioning

confidence: 99%

A truncated and dimeric format of an Affibody library on bacteria enables FACS‐mediated isolation of amyloid‐beta aggregation inhibitors with subnanomolar affinity

Lindberg

Härd

Löfblom

et al. 2015

Biotechnology Journal

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The amyloid hypothesis suggests that accumulation of amyloid b (Ab) peptides in the brain is involved in development of Alzheimer's disease. We previously generated a small dimeric affinity protein that inhibited Ab aggregation by sequestering the aggregation prone parts of the peptide. The affinity protein is originally based on the Affibody scaffold, but is evolved to a distinct interaction mechanism involving complex structural rearrangement in both the Ab peptide and the affinity proteins upon binding. The aim of this study was to decrease the size of the dimeric affinity protein and significantly improve its affinity for the Ab peptide to increase its potential as a future therapeutic agent. We combined a rational design approach with combinatorial protein engineering to generate two different affinity maturation libraries. The libraries were displayed on staphylococcal cells and high-affinity Ab-binding molecules were isolated using flow-cytometric sorting. The best performing candidate binds Ab with a K D value of around 300 pM, corresponding to a 50-fold improvement in affinity relative to the first-generation binder. The new dimeric Affibody molecule was shown to capture Ab 1-42 peptides from spiked E. coli lysate. Altogether, our results demonstrate successful engineering of this complex binder for increased affinity to the Ab peptide. Biotechnology JournalBiotechnol. J. 2015, 10, 1707-1718 therapies [8,16,17]. Alternative scaffold-proteins are in general much smaller than full-length antibodies and can usually be engineered into multivalent as well as multispecific units with an overall size that remain smaller than the conventional antibody [17,18]. Such features can potentially improve in vivo biodistribution of the agent [19][20][21][22]. One type alternative binding-protein that has been investigated for various diagnostic and therapeutic applications is the small three-helical bundle Affibody molecule (6.5 kDa) [17,23]. We previously generated an Affibody molecule (denoted Z Ab3 ) targeting monomeric Ab with a 17 nM affinity, as determined by isothermal titration calorimetry (ITC) and confirmed by surface plasmon resonance SPR [24][25][26]. Although this binder is based on the Affibody scaffold, it has evolved to adopt a unique and complex binding mechanism that is potentially more efficient for interactions with aggregation-prone peptides. Structural analysis has shown that two identical disulfide-linked Affibody units encapsulate one Ab peptide, and that both the Affibody domains and the Ab peptide undergo structural rearrangement upon binding. The Ab peptide folds into a b-hairpin structure, allowing the first α-helices in both Affibody molecules to adopt a b-sheet structure with unstructured N-termini [25,27]. In a recent in vivo study using an Ab-transgenic fruit fly model of AD, it was demonstrated that the Z Ab3 Affibody molecule efficiently inhibits the formation of Ab aggregates, thereby abolishing the neurotoxic effects and restoring the life span of the flies [25,28].For further po...

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“…Inside the brain, the antibody releases the receptor, and, with the other arm, binds and disrupts its main target: the enzyme betasecretase, which produces the amyloid-beta protein that accumulates in the brain in Alzheimer's disease. In a proof-of-concept study in 2014, the team showed that intravenous injections of the antibody lowered amyloidbeta levels in the brains of monkeys (4).…”

Section: Trojan Horsesmentioning

confidence: 99%

Circumventing the blood–brain barrier

Shen¹

2017

Proc. Natl. Acad. Sci. U.S.A.

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Therapeutic bispecific antibodies cross the blood-brain barrier in nonhuman primates

Cited by 296 publications

References 16 publications

Endosomal trafficking regulates receptor-mediated transcytosis of antibodies across the blood brain barrier

Endosomal trafficking regulates receptor-mediated transcytosis of antibodies across the blood brain barrier

A truncated and dimeric format of an Affibody library on bacteria enables FACS‐mediated isolation of amyloid‐beta aggregation inhibitors with subnanomolar affinity

Circumventing the blood–brain barrier

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