The objective of this R01 grant renewal is to characterize two novel transcriptional regulators, not previously implicated in stress or depression—and nearly unstudied in any neural phenomena—in mediating resilience to chronic stress via actions in the medial prefrontal cortex (mPFC) and other limbic brain regions. Virtually all prior studies of transcriptional mechanisms in stress action, including those supported by earlier editions of this grant, have taken a candidate approach. This is in marked contrast to an unbiased approach of using “big data” to deduce, in an open-ended manner, those factors that are most important in mediating specific aspects of stress susceptibility vs. resilience. We utilize this more powerful approach by taking advantage of large-scale RNA-seq datasets from mouse stress models and depressed humans to identify those factors that appear to play particularly critical roles in stress responses. We focus on cell type-specific actions of two of the most highly ranked transcriptional regulators from our datasets: ZFP189 (a deduced zinc finger transcription factor) and linc00473 (a primate- specific long non-coding RNA enriched in the cell nucleus). Zfp189 (ZNF189 in humans) is the top-ranked pro-resilience hub gene deduced in our datasets, while linc00473 is bioinformatically linked to ZFP189 in our human datasets and highly correlated with protein-coding genes that display abnormal expression in human depression. Interestingly, both appear to be downstream of the transcription factor, CREB. Based solely on these bioinformatics predictions, we have generated robust preliminary data to validate the importance of each of these transcriptional regulators in mouse stress models and now propose to better understand their actions. We will characterize their role in mPFC in controlling behavioral responses to chronic stress as well as map their target genes on a genome-wide basis, focusing on actions in pyramidal neurons where the two factors predominate. Related work will study the effect of ZFP189 and linc00473 on the excitability of this neuronal cell type in brain slices and in vivo. Together, this work will reveal new transcriptional mechanisms underlying behavioral resilience and inform our understanding of depression pathophysiology.