235 The importance of sirtuins in skin and new findings about sirt2 and its link to mechanobiology

Abstract

Sirtuins (SIRTs) are a family of NAD+-dependent histone deacetylases that play a critical role in many diverse cellular processes including transcriptional signaling, gene silencing, metabolism, genomic stability, inflammation, energy, stress response and aging. The objectives of this study were to define the temporal role of SIRTs in skin by kinetically quantifying SIRT expression and to assess environmental impact by examining the response to environmental stressors via measuring the effect on energy (ATP) production and oxidative damage (ROS). Additionally, the effect on mechanical properties such as collagen production were measured. Finally, we demonstrate the importance of SIRT2 and its effect on age-induced changes in cell shape. Sirtuin expression levels by Normal Human Epidermal Keratinocytes (NHEK) were measured by RT-PCR. NHEK were exposed to low doses of UVB (10mJ/cm2). Cellular energy (ATP) production as well as oxidative damage (H2O2) were measured. Finally, the effects of SIRTs on pro-collagen type I production in Normal Human Dermal Fibroblasts (NHDF) and cell spreading by aged NHDF were measured. Temporal differences in sirtuin expression levels were observed over time. Furthermore, SIRTs were impacted by environmental stressors such as UVB exposure and ozone, resulting in differences in ATP and H2O2 production. These data show that SIRTs 1,2,3 and 6 support cellular activity necessary for skin mechanical properties such as boosting collagen production and through SIRT-2 increasing cellular-spreading in aging skin cells. SIRTs exhibit temporal expression and are sensitive to environmental stressors. We show that SIRTs are beneficial for both cellular integrity and repair activity. Therefore, supporting their activity is imperative for maintaining optimal skin cell activity.