title: Suppression of the senescence-associated secretory phenotype (SASP) in human fibroblasts using small molecule inhibitors of p38 MAP kinase and MK2.
abstract: Senescent cells show an altered secretome profile termed the senescence-associated secretory phenotype (SASP). There is an increasing body of evidence that suggests that the accumulation of SASP-positive senescent cells in humans is partially causal in the observed shift to a low-level pro-inflammatory state in aged individuals. This in turn suggests the SASP as a possible therapeutic target to ameliorate inflammatory conditions in the elderly, and thus a better understanding of the signalling pathways underlying the SASP are required. Prior studies using the early generation p38 MAPK inhibitor SB203580 indicated that p38 signalling was required for the SASP. In this study, we extend these observations using two next-generation p38 inhibitors (UR-13756 and BIRB 796) that have markedly improved selectivity and specificity compared to SB203580, to strengthen the evidence that the SASP is p38-dependent in human fibroblasts. BIRB 796 has an efficacy and toxicity profile that has allowed it to reach Phase III clinical trials, suggesting its possible use to suppress the SASP in vivo. We also demonstrate for the first time a requirement for signalling through the p38 downstream MK2 kinase in the regulation of the SASP using two MK2 inhibitors. Finally, we demonstrate that a commercially-available multiplex cytokine assay technology can be used to detect SASP components in the conditioned medium of cultured fibroblasts from both young and elderly donors. This assay is a high-throughput, multiplex microtitre-based assay system that is highly sensitive, with very low sample requirements, allowing it to be used for low-volume human biological fluids. Our initial studies using existing multiplex plates form the basis for a "SASP signature" assay that could be used as a high-throughput system in a clinical study setting. Our findings therefore provide important steps towards the study of, and intervention in, the SASP in human ageing and age-related disease.
authors.affiliation: National Laboratory Astana, Centre for Life Sciences, Nazarbayev University, Astana, Kazakhstan.|School of Medicine, Institute of Cancer and Genetics, Cardiff University, Heath Park, Cardiff, CF14 4XN, UK.
authors.forename: Dauren
authors.initials: D
authors.lastname: Alimbetov
authors.affiliation: School of Medicine, Institute of Cancer and Genetics, Cardiff University, Heath Park, Cardiff, CF14 4XN, UK. davist2@cardiff.ac.uk.
authors.forename: Terence
authors.initials: T
authors.lastname: Davis
authors.affiliation: School of Medicine, Institute of Cancer and Genetics, Cardiff University, Heath Park, Cardiff, CF14 4XN, UK.
authors.forename: Amy J C
authors.initials: AJ
authors.lastname: Brook
authors.affiliation: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors.forename: Lynne S
authors.initials: LS
authors.lastname: Cox
authors.affiliation: School of Pharmacy and Biomolecular Sciences, University of Brighton, Cockcroft Building, Lewes Road, Moulsecoomb, Brighton, BN2 4GJ, UK.
authors.forename: Richard G A
authors.initials: RG
authors.lastname: Faragher
authors.affiliation: National Laboratory Astana, Centre for Life Sciences, Nazarbayev University, Astana, Kazakhstan.
authors.forename: Talgat
authors.initials: T
authors.lastname: Nurgozhin
authors.affiliation: National Laboratory Astana, Centre for Life Sciences, Nazarbayev University, Astana, Kazakhstan.
authors.forename: Zhaxybay
authors.initials: Z
authors.lastname: Zhumadilov
authors.affiliation: School of Medicine, Institute of Cancer and Genetics, Cardiff University, Heath Park, Cardiff, CF14 4XN, UK.
authors.forename: David
authors.initials: D
authors.lastname: Kipling
mesh_terms: D002478:Cells, Cultured; D016922:Cellular Senescence; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D011506:Proteins / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors*; D000099190:MAP-Kinase-Activated Kinase 2
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011506:Proteins; D048051:p38 Mitogen-Activated Protein Kinases; D000099190:MAP-Kinase-Activated Kinase 2
keywords: BIRB 796; Cellular senescence; Human ageing; Human fibroblasts; MK2.III; SB203580