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Exp Cell Res. 2016 Mar 15;342(2):145-58. doi: 10.1016/j.yexcr.2016.03.003. Epub 2016 Mar 9.

Targeting the replisome with transduced monoclonal antibodies triggers lethal DNA replication stress in cancer cells.

Author information

1
Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France.
2
Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/Université de Strasbourg, INSERM U964, rue Laurent Fries, 67404 Illkirch, France.
3
Faculté de Pharmacie, UMR 7213, CNRS/Université de Strasbourg, route du Rhin, 67401 Illkirch, France.
4
Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch, France. Electronic address: eweiss@unistra.fr.

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.

KEYWORDS:

Antibody transduction; Chemo-resistant cancer cells; DNA polymerase alpha; DNA replication stress; Functional inhibition; Proliferating cell nuclear antigen

PMID:
26968636
DOI:
10.1016/j.yexcr.2016.03.003
[Indexed for MEDLINE]

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