Transferrin receptor (TfR) trafficking determines brain uptake of TfR antibody affinity variants

Bien-Ly, Nga; Yu, Yang; Bumbaca, Daniela; Elstrott, Justin; Boswell, C. Andrew; Zhang, Yin; Luk, Wilman; Lu, Yanmei; Dennis, Mark S.; Weimer, Robby M.; Chung, Inhee; Watts, Ryan J.

doi:10.1084/jem.20131660

Cited by 260 publications

(268 citation statements)

References 25 publications

Supporting

Mentioning

247

Contrasting

Unclassified

Order By: Relevance

“…Consequently, the fraction of the antibody that actually gets through the BBB and into the brain parenchyma is very small. What has recently become evident in relation to BBB receptor engagement is the importance of understanding intracellular trafficking of the transporting receptor inside the brain endothelial cells and how its biology is influenced by the interaction with the carrier [13,14]. In agreement with earlier work our results show that the valency of the brain shuttle (i.e., number of modules interacting with the BBB receptor) is crucial for effective transport of the construct from the circulation into the brain parenchyma [13].…”

supporting

confidence: 90%

Time to Open the Blood–Brain Barrier Gate for Biologics?

Freskgård

Niewoehner²,

Urich

2014

Future Neurol.

View full text Add to dashboard Cite

supporting

confidence: 90%

Time to Open the Blood–Brain Barrier Gate for Biologics?

Freskgård

Niewoehner²,

Urich

2014

Future Neurol.

View full text Add to dashboard Cite

“…Multiple studies have shown that antibodies can promote internalization and lysosomal trafficking of their target receptors (38)(39)(40). In order to investigate the turnover of PRLR and HER2 in the absence of their respective antibodies, protein turnover in T47D cells was evaluated in a CHX chase assay.…”

Section: Prlr But Not Her2 Is Rapidly Degraded In Lysosomesmentioning

confidence: 99%

Bispecific Antibodies and Antibody–Drug Conjugates (ADCs) Bridging HER2 and Prolactin Receptor Improve Efficacy of HER2 ADCs

Andreev

Thambi

Bay

et al. 2017

Molecular Cancer Therapeutics

122

View full text Add to dashboard Cite

The properties of cell surface proteins targeted by antibodydrug conjugates (ADCs) have not been fully exploited; of particular importance are the rate of internalization and the route of intracellular trafficking. In this study, we compared the trafficking of HER2, which is the target of the clinically approved ADC ado-trastuzumab emtansine (T-DM1), with that of prolactin receptor (PRLR), another potential target in breast cancer. In contrast to HER2, we found that PRLR is rapidly and constitutively internalized, and traffics efficiently to lysosomes, where it is degraded. The PRLR cytoplasmic domain is necessary to promote rapid internalization and degradation, and when transferred to HER2, enhances HER2 degradation. In accordance with these findings, low levels of cell surface PRLR ($30,000 surface receptors per cell) are sufficient to mediate effective killing by PRLR ADC, whereas cell killing by HER2 ADC requires higher levels of cell surface HER2 ($10 6 surface receptors per cell). Noncovalently crosslinking HER2 to PRLR at the cell surface, using a bispecific antibody that binds to both receptors, dramatically enhances the degradation of HER2 as well as the cell killing activity of a noncompeting HER2 ADC. Furthermore, in breast cancer cells that coexpress HER2 and PRLR, a HER2xPRLR bispecific ADC kills more effectively than HER2 ADC. These results emphasize that intracellular trafficking of ADC targets is a key property for their activity and, further, that coupling an ADC target to a rapidly internalizing protein may be a useful approach to enhance internalization and cell killing activity of ADCs.

show abstract

“…11,13 Recent experience with antibodies against TfR as BBB-delivery carriers revealed that antibody characteristics, including affinity and valency, can dictate mechanisms of internalization, intracellular sorting and degradation in BEC and ultimately its transcytosis into the brain parenchyma. For example, decreasing antibody affinity 5,15 and using a mono-valent antiTfR antibody reduced intracellular degradation in BEC and enhanced transcytosis. 7 These modifications also improved the circulating half-life of bi-specific antibodies and enhanced the pharmacodynamic effect of therapeutic cargos, anti-BACE-1 5,15 and anti-b-amyloid 7 antibodies.…”

Section: Introductionmentioning

confidence: 99%

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

Haqqani

Delaney

Brunette

et al. 2017

J Cereb Blood Flow Metab

View full text Add to dashboard Cite

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.

show abstract

Transferrin receptor (TfR) trafficking determines brain uptake of TfR antibody affinity variants

Abstract: High-affinity transferrin receptor (TfR) bispecific antibodies facilitate trafficking of TfR to lysosomes and induce TfR degradation to decrease the ability of TfR to mediate BBB transcytosis.

Cited by 260 publications

References 25 publications

Time to Open the Blood–Brain Barrier Gate for Biologics?

Time to Open the Blood–Brain Barrier Gate for Biologics?

Bispecific Antibodies and Antibody–Drug Conjugates (ADCs) Bridging HER2 and Prolactin Receptor Improve Efficacy of HER2 ADCs

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

Contact Info

Product

Resources

About