title: Light-Controlled Delivery of Monoclonal Antibodies for Targeted Photoinactivation of Ki-67.
abstract: The selective inhibition of intracellular and nuclear molecules such as Ki-67 holds great promise for the treatment of cancer and other diseases. However, the choice of the target protein and the intracellular delivery of the functional agent remain crucial challenges. Main hurdles are (a) an effective delivery into cells, (b) endosomal escape of the delivered agents, and (c) an effective, externally triggered destruction of cells. Here we show a light-controlled two-step approach for selective cellular delivery and cell elimination of proliferating cells. Three different cell-penetrating nano constructs, including liposomes, conjugates with the nuclear localization sequence (NLS), and conjugates with the cell penetrating peptide Pep-1, delivered the light activatable antibody conjugate TuBB-9-FITC, which targets the proliferation associated protein Ki-67. HeLa cells were treated with the photosensitizer benzoporphyrin monoacid derivative (BPD) and the antibody constructs. In the first optically controlled step, activation of BPD at 690 nm triggered a controlled endosomal escape of the TuBB-9-FITC constructs. In more than 75% of Ki-67 positive, irradiated cells TuBB-9-FITC antibodies relocated within 24 h from cytoplasmic organelles to the cell nucleus and bound to Ki-67. After a second light irradiation at 490 nm, which activated FITC, cell viability decreased to approximately 13%. Our study shows an effective targeting strategy, which uses light-controlled endosomal escape and the light inactivation of Ki-67 for cell elimination. The fact that liposomal or peptide-assisted delivery give similar results leads to the additional conclusion that an effective mechanism for endosomal escape leaves greater variability for the choice of the delivery agent.
authors.affiliation: Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute of Biomedical Analytical Technology and Instrumentation, School of Life Science and Technology, Xi'an Jiaotong University , Xi'an 710049, P. R. China.|Institute of Biomedical Optics , University of Lübeck, Peter-Monnik-Weg 4, 23562 Lübeck, Germany.
authors.forename: Sijia
authors.initials: S
authors.lastname: Wang
authors.affiliation: Institute of Biomedical Optics , University of Lübeck, Peter-Monnik-Weg 4, 23562 Lübeck, Germany.
authors.forename: Gereon
authors.initials: G
authors.lastname: Hüttmann
authors.affiliation: Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute of Biomedical Analytical Technology and Instrumentation, School of Life Science and Technology, Xi'an Jiaotong University , Xi'an 710049, P. R. China.
authors.forename: Zhenxi
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Institute of Biomedical Optics , University of Lübeck, Peter-Monnik-Weg 4, 23562 Lübeck, Germany.
authors.forename: Alfred
authors.initials: A
authors.lastname: Vogel
authors.affiliation: Institute of Biomedical Optics , University of Lübeck, Peter-Monnik-Weg 4, 23562 Lübeck, Germany.
authors.forename: Reginald
authors.initials: R
authors.lastname: Birngruber
authors.affiliation: Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School , 50 Blossom Street, Boston, Massachusetts 02114, United States.
authors.forename: Shifalika
authors.initials: S
authors.lastname: Tangutoori
authors.affiliation: Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School , 50 Blossom Street, Boston, Massachusetts 02114, United States.
authors.forename: Tayyaba
authors.initials: T
authors.lastname: Hasan
authors.affiliation: Institute of Biomedical Optics , University of Lübeck, Peter-Monnik-Weg 4, 23562 Lübeck, Germany.
authors.forename: Ramtin
authors.initials: R
authors.lastname: Rahmanzadeh
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D003543:Cysteamine / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000737:chemistry; D011992:Endosomes / Q000378:metabolism; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D019394:Ki-67 Antigen / Q000737:chemistry* / Q000528:radiation effects*; D008027:Light; D008081:Liposomes / Q000737:chemistry*; D058990:Molecular Targeted Therapy; D019913:Nuclear Localization Signals; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000008:administration & dosage / Q000737:chemistry; D017319:Photosensitizing Agents / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D019394:Ki-67 Antigen; D008081:Liposomes; D019913:Nuclear Localization Signals; C503924:Pep-1 peptide; D010455:Peptides; D017319:Photosensitizing Agents; D003543:Cysteamine; D016650:Fluorescein-5-isothiocyanate
keywords: endosomal entrapment; liposome; nanotechnology; nuclear localization sequence (NLS); photodynamic therapy