Fig. 4: Synthetic ZFP189 TFs oppositely regulate transposable elements and genes related to immune function in prefrontal cortex.

A Twenty-four hours after viral delivery to prefrontal cortex (PFC), RNAseq was performed and transposable elements (TEs) were annotated in the reference library alongside differentially expressed genes (DEGs). On volcano plots, TEs are represented as larger dots and DEGs as smaller dots. In HSV-ZFP189^VPR manipulated PFC, hundreds of distinct TE metagenes are up-regulated (orange dots), with no downregulated TEs. Differential TEs and DEGs generated relative to HSV-ZFP189^NFD. Significance cutoff set to nominal p-value < .05. n = 5 microdissected PFC from individual mice (HSV-ZFP189^NFD), n = 5 microdissected PFC from individual mice (HSV-ZFP189^VPR). B In HSV-ZFP189^WT manipulated PFC, only one TE is detected as down-regulated relative to the control HSV-ZFP189^NFD (purple dot). Significance cutoff as in (A). n = 5 microdissected PFC from individual mice (HSV-ZFP189^NFD), n = 5 microdissected PFC from individual mice (HSV-ZFP189^WT). C Four days after viral delivery, another RNAseq was performed. Volcano plot depicting DEGs regulated by HSV-ZFP189^VPR PFC. Up-regulated DEGs are indicated by yellow dots, and down-regulated genes are indicated by blue dots. DEGs generated relative to our neutral HSV-ZFP189^NFD comparator. Significance cutoff set to 5% FDR corrected, Wald test adjusted p-value < 0.05. n = 7 microdissected PFC from individual mice (HSV-ZFP189^NFD), n = 8 microdissected PFC from individual mice (HSV-ZFP189^VPR). D Volcano plot depicting DEGs regulated by HSV-ZFP189^WT in PFC. DEGs generated relative to our neutral HSV-ZFP189^NFD comparator. Significance cutoff as in (B). n = 7 microdissected PFC from individual mice (HSV-ZFP189^NFD), n = 5 microdissected PFC from individual mice (HSV-ZFP189^WT). E Heatmap derived from Ingenuity Pathway Analysis (IPA) performed on complete DEGs lists from (C) and (D), weighted by fold change, reveals molecular factors whose function could explain the DEG expression profiles. HSV-ZFP189^WT vs. HSV-ZFP189^VPR regulated DEGs from (C) and (D) could largely be explained by increased (yellow) vs. decreased (blue) function of immune factors, respectively. F, G Gene ontology (GO) biological function terms enriched within the DEGs uniquely down-regulated by ZFP189^VPR (F) and from the DEGs uniquely up-regulated by ZFP189^WT (G) reveal that both DEGs lists are comprised of DEGs whose gene products participate in immune-related biological functions. H Cartoon representing how the normal function of ZFP189^WT can lead to the activation of immune-related transcripts and typical social behaviors. ZFP189^WT binds DNA motifs in TE-rich regions, represses the transcription of proximal TEs, recruits co-factors, and enables cis-regulatory action of this TE-rich region to enable the expression of immune genes in PFC, which, in turn, promotes social behaviors. Alternatively, ZFP189^VPR binds ZFP189 DNA motifs in TE-rich regions and activates the transcription of proximal TEs. This diminishes the cis-regulatory function of TE-rich regions and results in lowered expression of cis-regulated immune genes and impaired social behaviors. Cartoon made in Biorender.com.