Page 186 - ebook

[B. Cell Biology/Stem Cell] B-28

Discovering therapeutic agonists to stimulate TLR3-mediated

signaling in regeneration

Yukyung Park¹, Ji Yeon Moon¹, Woo Young Lee¹, MinJi Kim¹, Ga Ryung Kwon¹, Min A Ma¹,

Dongwon Kim¹*

¹Bio-chemical Engineering, Dongseo University, Busan 47011, Korea

Toll-Like-Receptor 3 (TLR3) induces double stranded RNAs (dsRNAs)-mediated signaling and retinoic acid synthesis

in skin regeneration.1,2 Although Poly I:C is a potent agonist to stimulate TLR3, it has structurally been unidentified
and severe cytotoxicity, suggesting that it may be hard to apply for human subjects. Given the regenerative

capacities in skin, discovering agonists on TLR3 activation may provide therapeutic mechanisms and develop
potential drugs in skin regeneration. To discover new molecules that could stimulate TLR3, we were provided with

6,696 chemical compounds randomly selected from Korea Chemical Bank. We cultured genetically modified HEK-
293 cells with TLR3-NFkB promoter which can be monitored colorimetric activity of secreted embryonic alkaline

phosphatase (SEAP). HEK cells were treated by 25 µM of each compound and 1 µg/ml of PIC as a positive control
for 18 hours. Optical densities measured at 655 nm wavelength were used to estimate effects on TLR3 activation,

compared to non-treated control groups. Finally, we selected 24 lead compounds and are currently testing them in
human neonatal foreskin keratinocytes using qRT-PCR and western blot analysis to confirm the effects of TLR3-

mediated signaling pathways. Nelson et al. (2016). Cell Stem Cell. 17(2):139-151 Kim et al. (2019). Nat Comm.
10:2811