title: Targeting the replisome with transduced monoclonal antibodies triggers lethal DNA replication stress in cancer cells.
abstract: Although chemical inhibition of the DNA damage response (DDR) in cancer cells triggers cell death, it is not clear if the fork blockade achieved with inhibitors that neutralise proteins of the replisome is sufficient on its own to overcome the DDR. Monoclonal antibodies to PCNA, which block the DNA elongation process in vitro, have been developed. When these antibodies were transduced into cancer cells, they are able to inhibit the incorporation of nucleoside analogues. When co-delivered with anti-PCNA siRNA, the cells were flattened and the size of their nuclei increased by up to 3-fold, prior to cell death. Analysis of these nuclei by super-resolution microscopy revealed the presence of large numbers of phosphorylated histone H2AX foci. A senescence-like phenotype of the transduced cells was also observed upon delivery of the corresponding Fab molecules or following PCNA gene disruption or when the Fab fragment of an antibody that neutralises DNA polymerase alpha was used. Primary melanoma cells and leukaemia cells that are resistant to chemical inhibitors were similarly affected by these antibody treatments. These results demonstrate that transduced antibodies can trigger a lethal DNA replication stress, which kills cancer cells by abolishing the biological activity of several constituents of the replisome.
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Desplancq
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Freund
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/Université de Strasbourg, INSERM U964, rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Sascha
authors.initials: S
authors.lastname: Conic
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Annie-Paule
authors.initials: AP
authors.lastname: Sibler
authors.affiliation: Faculté de Pharmacie, UMR 7213, CNRS/Université de Strasbourg, route du Rhin, 67401 Illkirch, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Didier
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Stoessel
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/Université de Strasbourg, INSERM U964, rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Mustapha
authors.initials: M
authors.lastname: Oulad-Abdelghani
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Vigneron
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Wagner
authors.affiliation: Faculté de Pharmacie, UMR 7213, CNRS/Université de Strasbourg, route du Rhin, 67401 Illkirch, France.
authors.forename: Yves
authors.initials: Y
authors.lastname: Mély
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Chatton
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/Université de Strasbourg, INSERM U964, rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Tora
authors.affiliation: Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France. Electronic address: eweiss@unistra.fr.
authors.forename: Etienne
authors.initials: E
authors.lastname: Weiss
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D053903:DNA Breaks, Double-Stranded; D004258:DNA Polymerase III / Q000037:antagonists & inhibitors; D004261:DNA Replication / Q000187:drug effects*; D004273:DNA, Neoplasm / Q000235:genetics* / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D004354:Drug Screening Assays, Antitumor; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D008807:Mice, Inbred BALB C; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013312:Stress, Physiological
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; C468783:H2AX protein, human; D006657:Histones; D007140:Immunoglobulin Fab Fragments; D018809:Proliferating Cell Nuclear Antigen; C065639:DNA polymerase A; D004258:DNA Polymerase III
keywords: Antibody transduction; Chemo-resistant cancer cells; DNA polymerase alpha; DNA replication stress; Functional inhibition; Proliferating cell nuclear antigen