title: Defective DSB repair correlates with abnormal nuclear morphology and is improved with FTI treatment in Hutchinson-Gilford progeria syndrome fibroblasts.
abstract: Impaired DSB repair has been implicated as a molecular mechanism contributing to the accelerating aging phenotype in Hutchinson-Gilford progeria syndrome (HGPS), but neither the extent nor the cause of the repair deficiency has been fully elucidated. Here we perform a quantitative analysis of the steady-state number of DSBs and the repair kinetics of ionizing radiation (IR)-induced DSBs in HGPS cells. We report an elevated steady-state number of DSBs and impaired repair of IR-induced DSBs, both of which correlated strongly with abnormal nuclear morphology. We recreated the HGPS cellular phenotype in human coronary artery endothelial cells for the first time by lentiviral transduction of GFP-progerin, which also resulted in impaired repair of IR-induced DSBs, and which correlated with abnormal nuclear morphology. Farnesyl transferase inhibitor (FTI) treatment improved the repair of IR-induced DSBs, but only in HGPS cells whose nuclear morphology was also normalized. Interestingly, FTI treatment did not result in a statistically significant reduction in the higher steady-state number of DSBs. We also report a delay in localization of phospho-NBS1 and MRE11, MRN complex repair factors necessary for homologous recombination (HR) repair, to DSBs in HGPS cells. Our results demonstrate a correlation between nuclear structural abnormalities and the DSB repair defect, suggesting a mechanistic link that may involve delayed repair factor localization to DNA damage. Further, our results show that similar to other HGPS phenotypes, FTI treatment has a beneficial effect on DSB repair.
authors.affiliation: Department of Cell Biology-Physiology, University of Pittsburgh, Pittsburgh, PA 15260, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Constantinescu
authors.forename: Antonei B
authors.initials: AB
authors.lastname: Csoka
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Navara
authors.forename: Gerald P
authors.initials: GP
authors.lastname: Schatten
mesh_terms: D016022:Case-Control Studies; D002467:Cell Nucleus / Q000473:pathology*; D002478:Cells, Cultured; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D051231:Farnesyltranstransferase / Q000037:antagonists & inhibitors*; D005347:Fibroblasts / Q000187:drug effects / Q000473:pathology*; D006801:Humans; D011371:Progeria / Q000188:drug therapy / Q000473:pathology*; D013577:Syndrome
chemical_list: D004791:Enzyme Inhibitors; D051231:Farnesyltranstransferase