title: Receptor Crosslinking: A General Method to Trigger Internalization and Lysosomal Targeting of Therapeutic Receptor:Ligand Complexes.
abstract: A major unmet clinical need is a universal method for subcellular targeting of bioactive molecules to lysosomes. Delivery to this organelle enables either degradation of oncogenic receptors that are overexpressed in cancers, or release of prodrugs from antibody-drug conjugates. Here, we describe a general method that uses receptor crosslinking to trigger endocytosis and subsequently redirect trafficking of receptor:cargo complexes from their expected route, to lysosomes. By incubation of plasma membrane receptors with biotinylated cargo and subsequent addition of streptavidin to crosslink receptor:cargo-biotin complexes, we achieved rapid and selective lysosomal targeting of transferrin, an anti-MHC class I antibody, and the clinically approved anti-Her2 antibody trastuzumab. These three protein ligands each target a receptor with a distinct cellular function and intracellular trafficking profile. Importantly, we confirmed that crosslinking of trastuzumab increased lysosomal degradation of its cognate oncogenic receptor Her2 in breast cancer cell lines SKBR3 and BT474. These data suggest that crosslinking could be exploited for a wide range of target receptors, for navigating therapeutics through the endolysosomal pathway, for significant therapeutic benefit.
authors.affiliation: Cardiff School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff, Wales.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Moody
authors.affiliation: Cardiff School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff, Wales.
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Sayers
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, England.
authors.forename: Johannes P
authors.initials: JP
authors.lastname: Magnusson
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, England.
authors.forename: Cameron
authors.initials: C
authors.lastname: Alexander
authors.affiliation: School of Biosciences, Cardiff University, Cardiff, Wales.
authors.forename: Paola
authors.initials: P
authors.lastname: Borri
authors.affiliation: School of Biosciences, Cardiff University, Cardiff, Wales.
authors.forename: Peter
authors.initials: P
authors.lastname: Watson
authors.affiliation: Cardiff School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff, Wales.
authors.forename: Arwyn T
authors.initials: AT
authors.lastname: Jones
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods*; D004705:Endocytosis / Q000187:drug effects; D005260:Female; D018390:Gene Targeting / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D018796:Immunoconjugates / Q000494:pharmacology; D008024:Ligands; D008247:Lysosomes / Q000378:metabolism*; D011355:Prodrugs; D021381:Protein Transport; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D000068878:Trastuzumab / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D018796:Immunoconjugates; D008024:Ligands; D011355:Prodrugs; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human