Transcytosis of payloads that are non-covalently complexed to bispecific antibodies across the hCMEC/D3 blood-brain barrier model
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Daniela Schmid
Abstract
A transcellular shuttle system was generated for the delivery of non-covalently linked payloads across blood-brain barrier (BBB) endothelial cells. Transcytosis-enabling shuttles are composed of bispecific antibodies (bsAbs) that simultaneously bind transferrin receptor (TfR) and haptens such as digoxigenin or biocytinamide. Haptenylated payloads are attached to these vehicles via non-covalent hapten-antibody complexation. This enables targeting to and internalization into human BBB-derived microvascular endothelial hCMEC/D3 cells. In contrast to other shuttles, this system does not require special affinities or formats of their TfR-binding moieties for transcytosis and subsequent release. Non-covalent payload complexation to bsAb is flexible and robust, works for a multitude of payloads and enables separation of payloads from shuttles during transcytosis. Released payloads can enter the brain without connected bsAb entities, minimizing potential interference with distribution or functionality. Intracellular separation of shuttle and payload and recycling to cell surfaces may also enable recharging of the cell-bound BBB shuttle with payload for subsequent (merry-go-round) transport cycles.
Acknowledgements
We thank Heike Seul and Martina Wagner for excellent support and contributions. D.S. and A.B. are supported by the Roche Postdoc Fund (RPF)/Targeted Therapies.
Conflict of interest statement: The authors are employees of Roche Pharma Research and Early Development. Roche is interested in targeted therapies.
References
Banks, W.A. (2016). From blood-brain barrier to blood-brain interface: new opportunities for CNS drug delivery. Nat. Rev. Drug Discov. 15, 275–292.10.1038/nrd.2015.21Suche in Google Scholar PubMed
Bera, T.K., Onda, M., Brinkmann, U., and Pastan, I. (1998). A bivalent disulfide-stabilized Fv with improved antigen binding to erbB2. J. Mol. Biol. 281, 475–483.10.1006/jmbi.1998.1948Suche in Google Scholar PubMed
Brinkmann, U. and Kontermann, R.E. (2017). The making of bispecific antibodies. MAbs 9, 182–212.10.1080/19420862.2016.1268307Suche in Google Scholar PubMed PubMed Central
Brinkmann, U., Reiter, Y., Jung, S.H., Lee, B., and Pastan, I. (1993). A recombinant immunotoxin containing a disulfide-stabilized Fv fragment. Proc. Natl. Acad. Sci. USA 90, 7538–7542.10.1073/pnas.90.16.7538Suche in Google Scholar PubMed PubMed Central
Dengl, S., Hoffmann, E., Grote, M., Wagner, C., Mundigl, O., Georges, G., Thorey, I., Stubenrauch, K.G., Bujotzek, A., Josel, H.P., et al. (2015). Hapten-directed spontaneous disulfide shuffling: a universal technology for site-directed covalent coupling of payloads to antibodies. FASEB J. 29, 1763–1779.10.1096/fj.14-263665Suche in Google Scholar PubMed PubMed Central
Dengl, S., Sustmann, C., and Brinkmann, U. (2016). Engineered hapten-binding antibody derivatives for modulation of pharmacokinetic properties of small molecules and targeted payload delivery. Immunol. Rev. 270, 165–177.10.1111/imr.12386Suche in Google Scholar PubMed PubMed Central
Evers, M.M., Toonen, L.J., and van Roon-Mom, W.M. (2015). Antisense oligonucleotides in therapy for neurodegenerative disorders. Adv. Drug Deliv. Rev. 87, 90–103.10.1016/j.addr.2015.03.008Suche in Google Scholar PubMed
Friden, P.M., Olson, T.S., Obar, R., Walus, L.R., and Putney, S.D. (1996). Characterization, receptor mapping and blood-brain barrier transcytosis of antibodies to the human transferrin receptor. J. Pharmacol. Exp. Ther. 278, 1491–1498.10.1016/S0022-3565(25)20804-7Suche in Google Scholar
Grote, M., Haas, A.K., Klein, C., Schaefer, W., and Brinkmann, U. (2012). Bispecific antibody derivatives based on full-length IgG formats. Methods Mol. Biol. 901, 247–263.10.1007/978-1-61779-931-0_16Suche in Google Scholar PubMed
Hoffmann, E., Konkar, A., Dziadek, S., Josel, H.P., Conde-Knape, K., Kropp, H., Kling, L., Stubenrauch, K., Thorey, I., Dengl, S., et al. (2013). PK modulation of haptenylated peptides via non-covalent antibody complexation. J. Contr. Release 171, 48–56.10.1016/j.jconrel.2013.06.021Suche in Google Scholar PubMed
Kontermann, R.E. and Brinkmann, U. (2015). Bispecific antibodies. Drug Discov. Today 20, 838–847.10.1016/j.drudis.2015.02.008Suche in Google Scholar PubMed
Metz, S., Haas, A.K., Daub, K., Croasdale, R., Stracke, J., Lau, W., Georges, G., Josel, H.P., Dziadek, S., Hopfner, K.P., et al. (2011). Bispecific digoxigenin-binding antibodies for targeted payload delivery. Proc. Natl. Acad. Sci. USA 108, 8194–8199.10.1073/pnas.1018565108Suche in Google Scholar PubMed PubMed Central
Niewoehner, J., Bohrmann, B., Collin, L., Urich, E., Sade, H., Maier, P., Rueger, P., Stracke, J.O., Lau, W., Tissot, A.C., et al. (2014). Increased brain penetration and potency of a therapeutic antibody using a monovalent molecular shuttle. Neuron 81, 49–60.10.1016/j.neuron.2013.10.061Suche in Google Scholar PubMed
Pardridge, W.M. (2017). Delivery of biologics across the blood-brain barrier with molecular trojan horse technology. BioDrugs 31, 503–519.10.1007/s40259-017-0248-zSuche in Google Scholar PubMed
Pardridge, W.M. and Boado, R.J. (2012). Reengineering biopharmaceuticals for targeted delivery across the blood-brain barrier. Methods Enzymol. 503, 269–292.10.1016/B978-0-12-396962-0.00011-2Suche in Google Scholar PubMed
Reiter, Y., Brinkmann, U., Kreitman, R.J., Jung, S.H., Lee, B., and Pastan, I. (1994). Stabilization of the Fv fragments in recombinant immunotoxins by disulfide bonds engineered into conserved framework regions. Biochemistry 33, 5451–5459.10.1021/bi00184a014Suche in Google Scholar PubMed
Reiter, Y., Brinkmann, U., Jung, S.H., Pastan, I., and Lee, B. (1995). Disulfide stabilization of antibody Fv: computer predictions and experimental evaluation. Protein Eng. 8, 1323–1331.10.1093/protein/8.12.1323Suche in Google Scholar PubMed
Sade, H., Baumgartner, C., Hugenmatter, A., Moessner, E., Freskgard, P.O., and Niewoehner, J. (2014). A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. PLoS One 9, e96340.10.1371/journal.pone.0096340Suche in Google Scholar PubMed PubMed Central
Schneider, B., Grote, M., John, M., Haas, A., Bramlage, B., Ickenstein, L.M., Jahn-Hofmann, K., Bauss, F., Cheng, W., Croasdale, R., et al. (2012). Targeted siRNA delivery and mRNA knockdown mediated by bispecific digoxigenin-binding antibodies. Mol. Ther. Nucl. Acids 1, e46.10.1038/mtna.2012.39Suche in Google Scholar PubMed PubMed Central
Thorey, I.S., Grote, M., Mayer, K., and Brinkmann, U. (2016). Hapten-binding bispecific antibodies for the targeted delivery of siRNA and siRNA-containing nanoparticles. Methods Mol. Biol. 1364, 219–234.10.1007/978-1-4939-3112-5_18Suche in Google Scholar PubMed
Weksler, B.B., Subileau, E.A., Perriere, N., Charneau, P., Holloway, K., Leveque, M., Tricoire-Leignel, H., Nicotra, A., Bourdoulous, S., Turowski, P., et al. (2005). Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J. 19, 1872–1874.10.1096/fj.04-3458fjeSuche in Google Scholar PubMed
Weksler, B., Romero, I.A., and Couraud, P.O. (2013). The hCMEC/D3 cell line as a model of the human blood brain barrier. Fluids Barriers CNS 10, 16.10.1186/2045-8118-10-16Suche in Google Scholar PubMed PubMed Central
Yu, Y.J., Zhang, Y., Kenrick, M., Hoyte, K., Luk, W., Lu, Y., Atwal, J.K., Elliott, J.M., Prabhu, S., Watts, R.J., et al. (2011). Boosting brain uptake of a therapeutic antibody by reducing its affinity for a transcytosis target. Sci. Transl. Med. 3, 84ra44.10.1126/scitranslmed.3002230Suche in Google Scholar PubMed
Yu, Y.J., Atwal, J.K., Zhang, Y., Tong, R.K., Wildsmith, K.R., Tan, C., Bien-Ly, N., Hersom, M., Maloney, J.A., Meilandt, W.J., et al. (2014). Therapeutic bispecific antibodies cross the blood-brain barrier in nonhuman primates. Sci. Transl. Med. 6, 261ra154.10.1126/scitranslmed.3009835Suche in Google Scholar PubMed
Supplemental Material:
The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2017-0311).
©2018 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Highlight Issue ‘Molecular Basis of Life 2017’
- HIGHLIGHT: GBM Fall Meeting “Molecular Basis of Life 2017”
- Neuronal RNP granules: from physiological to pathological assemblies
- Regulation of LRRK2: insights from structural and biochemical analysis
- The role of (auto)-phosphorylation in the complex activation mechanism of LRRK2
- Oncogenic BRAFV600E drives expression of MGL ligands in the colorectal cancer cell line HT29 through N-acetylgalactosamine-transferase 3
- Hypoxia and serum deprivation induces glycan alterations in triple negative breast cancer cells
- Targeting autophagy for the treatment of cancer
- From molecules to patients: exploring the therapeutic role of soluble guanylate cyclase stimulators
- DNA-encoded libraries – an efficient small molecule discovery technology for the biomedical sciences
- Transcytosis of payloads that are non-covalently complexed to bispecific antibodies across the hCMEC/D3 blood-brain barrier model
- Mitochondrial contributions to neuronal development and function
- Intracellular communication between lipid droplets and peroxisomes: the Janus face of PEX19
- Protein crystallization in living cells
- Synthetic DNA filaments: from design to applications
- Spectroscopic characterization of the Co-substituted C-terminal domain of rubredoxin-2
- Twitch or swim: towards the understanding of prokaryotic motion based on the type IV pilus blueprint
Artikel in diesem Heft
- Frontmatter
- Highlight Issue ‘Molecular Basis of Life 2017’
- HIGHLIGHT: GBM Fall Meeting “Molecular Basis of Life 2017”
- Neuronal RNP granules: from physiological to pathological assemblies
- Regulation of LRRK2: insights from structural and biochemical analysis
- The role of (auto)-phosphorylation in the complex activation mechanism of LRRK2
- Oncogenic BRAFV600E drives expression of MGL ligands in the colorectal cancer cell line HT29 through N-acetylgalactosamine-transferase 3
- Hypoxia and serum deprivation induces glycan alterations in triple negative breast cancer cells
- Targeting autophagy for the treatment of cancer
- From molecules to patients: exploring the therapeutic role of soluble guanylate cyclase stimulators
- DNA-encoded libraries – an efficient small molecule discovery technology for the biomedical sciences
- Transcytosis of payloads that are non-covalently complexed to bispecific antibodies across the hCMEC/D3 blood-brain barrier model
- Mitochondrial contributions to neuronal development and function
- Intracellular communication between lipid droplets and peroxisomes: the Janus face of PEX19
- Protein crystallization in living cells
- Synthetic DNA filaments: from design to applications
- Spectroscopic characterization of the Co-substituted C-terminal domain of rubredoxin-2
- Twitch or swim: towards the understanding of prokaryotic motion based on the type IV pilus blueprint
