492 Bi-layered skin construct with gene modified keratinocytes reduces UVR DNA damage

Ultraviolet radiation (UVR) is among major contributors to skin cancer in humans. Production of melanin serves as a main defense mechanisms against UVR damage. This pigment, which gives color hair, and eyes, protects the underlying tissue from However, some patient populations, this protection mechanism not effective, leaving patients highly susceptible cancer. Similarly, mycosporine like amino acids (MAAs), are commonly expressed marine algae, have been shown serve protective role UVR-induced DNA Cyanobacterium Cylindrospermum stagnale has gene cluster five genes (mylA-mylE) for MAA biosynthesis. The purpose study determine if human cells engineered express situ will provide UVR. MylA-mylE (MAA) was cloned into lentivirus vector used transduce dermal keratinocytes. Subsequently, bi-layered constructs with MAA-expressing, recombinant keratinocytes fibroblasts were fabricated fibrinogen hydrogel. exposed UVB over four days, then maintained 11 days culture media. Live/dead staining demonstrated higher cell viability construct compared normal controls. There also reduced TUNEL activity, active caspase 3, double strand breaks, γH2AX levels these constructs, Taken together, demonstrates that producing increased an associated reduction damage cellular death. technology may be utilized preferred method generation innate high susceptibility