In aging, unresolved cellular stress leads to chronic upregulation of cell stress signaling pathways.  In cell stress signaling, we observed impaired inflammatory signaling by measuring mRNA of PTGS2/Cox2, IL-6, TNFα, IL-1β, upregulated from the NF-κB transcription factor (ROS/NF-κB and JNK/NF-κB) and observed chronic activation of ER stress by measuring mRNA expression of CHOP (C/EBP-homologous protein, a transcription factor implicated in the regulation of apoptosis) and Ern1.  We also analyzed cell cycle regulators and the DNA damage response pathway and observed downregulation of ATM, upregulation of p21, XBP1 and caspase3 and 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.  

Aging phenotypes also exhibit down-regulation of cytoprotective pathways.  We saw downregulation of the pro-survival pathway (BCL2) and impaired activation of the Nrf2 transcription factor, leading to constitutive activation of HMOX1 and GCLC/GCLM genes. 

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