title: AvidinOX-anchored biotinylated trastuzumab and pertuzumab induce down-modulation of ErbB2 and tumor cell death at concentrations order of magnitude lower than not-anchored antibodies.
abstract: The oxidized version of Avidin, known as AvidinOX, was previously shown to link to tissue proteins upon injection or nebulization, thus becoming a stable receptor for biotinylated therapeutics. AvidinOX is currently under clinical investigation to target radioactive biotin to inoperable tumor lesions (ClinicalTrials.gov NCT02053324). Presently, we show that the anti-ErbB2 monoclonal antibodies Trastuzumab and Pertuzumab can be chemically biotinylated while maintaining their biochemical and biological properties. By using several and diverse experimental conditions, we show that when AvidinOX is conjugated to tumor cells, low antibody concentrations of biotinylated Trastuzumab (bTrast) or Pertuzumab (bPert) prevent internalization of ErbB2, induce endoplasmic reticulum stress, cell cycle arrest and apoptosis leading to inhibition of proliferation and ErbB2 signaling. Moreover, we found that the treatment is able to induce down-modulation of ErbB2 thus bypassing the known resistance of this receptor to degradation. Interestingly, we show that AvidinOX anchorage is a way to counteract agonistic activities of Trastuzumab and Pertuzumab. Present data are in agreement with previous observations from our group indicating that the engagement of the Epidermal Growth Factor Receptor (EGFR) by AvidinOX-bound biotinylated Cetuximab or Panitumumab, leads to potent tumor inhibition both in vitro and in animal models. All results taken together encourage further investigation of AvidinOX-based treatments with biotinylated antibodies directed to the members of the EGFR family.
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Ferdinando Maria
authors.initials: FM
authors.lastname: Milazzo
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Anastasi
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Chiapparino
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Rosi
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Leoni
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Vesci
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Fiorella
authors.initials: F
authors.lastname: Petronzelli
authors.affiliation: Biotech Products, Research and Development, Sigma-Tau SpA, 00071 Pomezia (Rome), Italy.
authors.forename: Rita
authors.initials: R
authors.lastname: De Santis
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001360:Avidin / Q000737:chemistry / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism; D019921:Biotinylation; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D006801:Humans; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000276:immunology; D015398:Signal Transduction; D000068878:Trastuzumab / Q000008:administration & dosage; D014407:Tumor Cells, Cultured
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D001360:Avidin; D014408:Biomarkers, Tumor; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human; C485206:pertuzumab
keywords: ErbB2; avidinOX; cancer; pertuzumab; trastuzumab