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

J Exp Med. 2014 Feb 10;211(2):233-44. doi: 10.1084/jem.20131660. Epub 2014 Jan 27.

Abstract

Antibodies to transferrin receptor (TfR) have potential use for therapeutic entry into the brain. We have shown that bispecific antibodies against TfR and β-secretase (BACE1 [β-amyloid cleaving enzyme-1]) traverse the blood-brain barrier (BBB) and effectively reduce brain amyloid β levels. We found that optimizing anti-TfR affinity improves brain exposure and BACE1 inhibition. Here we probe the cellular basis of this improvement and explore whether TfR antibody affinity alters the intracellular trafficking of TfR. Comparing high- and low-affinity TfR bispecific antibodies in vivo, we found that high-affinity binding to TfR caused a dose-dependent reduction of brain TfR levels. In vitro live imaging and colocalization experiments revealed that high-affinity TfR bispecific antibodies facilitated the trafficking of TfR to lysosomes and thus induced the degradation of TfR, an observation which was further confirmed in vivo. Importantly, high-affinity anti-TfR dosing induced reductions in brain TfR levels, which significantly decreased brain exposure to a second dose of low-affinity anti-TfR bispecific. Thus, high-affinity anti-TfR alters TfR trafficking, which dramatically impacts the capacity for TfR to mediate BBB transcytosis.

MeSH terms

  • Amyloid Precursor Protein Secretases / immunology
  • Animals
  • Antibodies, Bispecific / metabolism*
  • Antibody Affinity
  • Aspartic Acid Endopeptidases / immunology
  • Blood-Brain Barrier / immunology
  • Blood-Brain Barrier / metabolism
  • Brain / immunology*
  • Brain / metabolism*
  • Female
  • Lysosomes / immunology
  • Lysosomes / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Protein Transport
  • Receptors, Transferrin / immunology*
  • Receptors, Transferrin / metabolism*
  • Transcytosis / immunology
  • Transcytosis / physiology

Substances

  • Antibodies, Bispecific
  • Receptors, Transferrin
  • Tfrc protein, mouse
  • Amyloid Precursor Protein Secretases
  • Aspartic Acid Endopeptidases
  • Bace1 protein, mouse