Epigenetic biomarkers of technology efficacy

Year 1/2: Develop predictive in-vitro epigenetic screening methodology, (low risk - all techniques running at Manchester):
a. Develop low cost qPCR methylation method for stress response DNAm panel.
b. Develop repeat in-vitro UV treatment of keratinocytes and confirm expected DNAm changes (via qPCR).
c. Investigate potential mechanisms of action (MoA) of DNAm changes using bioinformatic approaches.
Year 2: Test known anti-ageing compounds for protective effects in a human keratinocyte UV model from 1).
d. Select compounds for screening that that are in topical products, have a known anti-stress mechanism of action MoA, and/or modulate biological pathways linked to DNAm changes from 1c).
e. Test ~40 candidate compounds for protective effects to UV as measured by DNAm.
If no DNAm benefit, we will use whole genome DNAm & RNAseq to identify where benefits are occurring and why these might not be evident in the specifically selected DNAm markers. We will then evaluate alternative DNAm markers for 3). Additionally, 4) can still proceed.
f. Confirm UV protection MoA through measuring key protein changes identified in 2a) and cellular damage markers.
Year 3/4 Clinical proof-of-principle test of lead compound from 2) for anti-stress effects in-vivo, replicating previous BBSRC iCASE clinical (hence, low risk):
g. Recruit subjects.
h. Applying a placebo control and compound treatment on subject skin, alongside UV dosing - take skin biopsies.
i. Extract DNA and examine changes to DNAm markers using qPCR.
j. Confirm MoA of protection through measuring key protein changes and cellular damage markers from 2c).
Year 1-4 (i.e. flexible timing) Investigate changes that occur in the dermis from repeat UV radiation.
k. Extract DNA and RNA from previously collected dermal samples (hence, low risk).
l. Carry out whole genome DNAm and RNA analysis.
m. Identify UV sensitive DNAm and RNA markers, as well as young old differences in response. Compare with epidermal findings.