In aging, unresolved cellular stress leads to chronic upregulation of cell stress signaling pathways and down-regulation of cyto-protective pathways. In cell stress signaling, we observed impaired inflammatory signaling by measuring IL-6, TNFα, IL-1β and the transcription factor of NF-κB (PTGS2/Cox2) and observed chronic upregulation of ER stress by measuring CHOP (C/EBP-homologous protein, a transcription factor implicated in the regulation of apoptosis) and Ern1 expression. We also analyzed cell cycle regulators and DNA damage response pathway and observed downregulation of ATM, ATR, upregulation of p21, XBP1 and caspase3, as well as downregulation of the pro-survival pathway (BCL2) and upregulation of pro-apoptotic proteins; all pointing towards either cell cycle arrest or activation of apoptotic pathways in the face of a severe insult resulting from a state of chronic stress. In our studies, we also saw constitutive activation of HMOX1 and GCLC/GCLM with impaired activation of the Nrf2 transcription factor.

AgeisBio has identified a complex transcriptional network that includes these key regulatory genes. Modulation of our target rejuvenated the cells, pushing them towards cell survival phenotypes as evidenced by the reduction seen in cell stress markers and the up-regulation of cyto-protective proteins.