Patent Publication Number: US-2020291005-A1

Title: Compositions and methods for inhibiting acss2

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims priority to U.S. Provisional Application Ser. No. 62/563,148, filed Sep. 26, 2017, the content of which is incorporated by reference herein in its entirety. 
    
    
     STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT 
     This invention was made with government support under Grant No. P01AG031862 awarded by The National Institutes for Health. The government has certain rights in the invention. 
    
    
     BACKGROUND OF THE INVENTION 
     Memory formation involves synaptic restructuring and requires the coordinated expression of neuronal genes through poorly understood processes that modify chromatin (Kandel, E. R. et al., 2014, Cell, 157:163-186; Zovkic, I. B. et al., 2013, Learn. Mem., 20:61-74). Histone acetylation is a key regulator of memory storage and restructures chromatin in distinct brain regions that have been implicated in learning and memory, most prominently in the hippocampus (Graff, J. et al., 2013, Nat. Rev. Neurosci., 14:97-111). Hippocampal memory consolidation requires the transcription factor CREB and the coactivator CREB binding protein (CBP), specifically the histone acetyltransferase (HAT) activity of CBP (Wood, M. A. et al., 2005, Learn. Mem., 12:111-119; Korzus, E. et al., 2004, Neuron, 42:961-972). Furthermore, inhibitors of histone deacetylases enhance memory consolidation (Graff, J. et al., 2013, Nat. Rev. Neurosci., 14:97-111). However, the mechanisms that regulate neuronal histone acetylation in long-term memory remain incompletely understood. 
     Direct sensing of intermediary metabolites by chromatin-modifying enzymes such as acetyltransferases can dynamically adapt chromatin structure and gene expression (Kaelin, W. G. Jr. et al., 2013, Cell, 153:56-69; Katada, S., et al., 2012, Cell, 148:24-28). Alteration of pools of intracellular acetyl-CoA manipulates histone acetylation (Cai, L., et al., 2011, Mol. Cell, 42:426-437; Wellen, K. E. et al., 2009, Science, 324:1076-1080); thus, metabolic enzymes that generate nuclear acetyl-CoA might directly control histone acetylation and gene expression (Gut, P. et al., 2013, Nature, 502:489-498; Pietrocola, F. et al., 2015, Cell Metab., 21:805-821). In mammalian cells, there are two principal enzymes that generate acetyl-CoA for histone acetylation: acetate-dependent acetyl-CoA synthetase 2 (ACSS2) and citrate-dependent ATP-citrate lyase (ACL) (Pietrocola, F. et al., 2015, Cell Metab., 21:805-821). The relative importance of ACSS2 and ACL for nuclear histone acetylation differs by tissue type, developmental state, and disease (Wellen, K. E. et al., 2009, Science, 324:1076-1080; Pietrocola, F. et al., 2015, Cell Metab., 21:805-821), but the roles of these enzymes in post-mitotic neuronal cells are unknown. 
     Thus, there remains a need in the art for therapies to treat neurological and cognitive diseases and disorders. The present invention addresses this unmet need. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The following detailed description of embodiments of the invention will be better understood when read in conjunction with the appended drawings. It should be understood that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings. 
         FIG. 1 , comprising  FIG. 1A  through  FIG. 1G , depicts results from example experiments demonstrating nuclear ACSS2 supports neuronal gene expression. ( FIG. 1A ) ACSS2 localizes to the cytoplasm in undifferentiated CAD neurons. ACSS2 was imaged by immunofluorescence microscopy in CAD cells (4′,6-diamidino-2-phenylindole (DAPI) and α-tubulin (α-Tub) immunostaining show nuclei and cytoplasm, respectively). ( FIG. 1B ) ACSS2 localizes to the nucleus in differentiated CAD neurons. ( FIG. 1C ) Western blot analysis of cytoplasmic (CE) and nuclear (NE) extracts from undifferentiated CAD cells (undiff.) and differentiated CAD neurons (diff.) for ACSS2, ACL and histone H3. Nuclear ACSS2 expression is higher in differentiated cells (p.d.u., procedure defined unit; t-test P=0.002, n=3, mean±s.d.). ( FIG. 1D ) ACSS2 knockdown reduces differentiation-linked upregulation of neuronal gene expression program. Scatter plot contrasts the fold-change fragments per kilobase of transcript per million mapped (FPKM) of induced genes ( FIG. 6C ) between wild-type (WT) and ACSS2 knockdown (KD) cells. Marginal distributions show histogram and kernel density estimation. Ordinary least squares linear regression is displayed with 95% confidence interval. ( FIG. 1E ) Western blot of lysates from differentiated CAD neurons that were infected with lentiviral control (WT) or ACSS2 knockdown vector (shACSS2) (quantification shown in  FIG. 5G ; n=3). ( FIG. 1F ) ACSS2 knockdown greatly reduces gene upregulation. Quintiles of upregulated genes (red dots in  FIG. 6C ) with the greatest fold-change increase in wild-type cells (grey). Corresponding gene quintiles depict fold-change FPKM in ACSS2 knockdown cells (green) (for each quintile, columns represent the mean induction value of genes; mean±s.e.m.). ( FIG. 1G ) ACSS2i treatment of differentiated CAD neurons results in reduced expression of differentiation-induced genes. All genes are plotted in order of fold-change in wild-type CAD differentiation, and z-scores were computed for ACSS2i treatment and control, representing upregulation as blue and downregulation as red (RNA-seq in 24-hour ACSS2i treated and DMSO-treated control neurons, genes removed with z-score &lt;0.5). Scale bar, 10 m ( FIG. 1A ,  FIG. 1B ). 
         FIG. 2 , comprising  FIG. 2A  through  FIG. 2J , depicts results from example experiments demonstrating that ACSS2 is recruited to transcriptionally active chromatin and promotes neuronal histone acetylation. ( FIG. 2A ) Genome browser tracks showing ChIP-seq over the Camk2a locus show that increases in H4K5, H4K12, and H3K9 acetylation co-occur with proximate ACSS2 enrichment upon CAD neuron differentiation (chromosome 18: 60,920,000-60,990,000). ( FIG. 2B ) Gene ontology term enrichment analysis of top 5% genes that become ACSS2-bound during CAD neuron differentiation show neuronal pathways. ( FIG. 2C ) Violin-contour plots show that ChIP-seq enrichment of the indicated histone acetylation occurs with top-ranked ACSS2 enrichment during neuronal differentiation of CAD cells. ( FIG. 2D ) ChIP-seq enrichment of the 299 genes that are reduced upon ACSS2i treatment (see Example 1 Methods) shows high correlation (P&lt;2.2×10 −16  for all) with histone acetylation in the differentiated state (AUC, area under the curve; d, differentiated; u, undifferentiated). ( FIG. 2E ) Analysis of all genes previously linked to neuronal differentiation (ND genes, AmiGO annotation set of 1,315 genes), and the subset of known ND genes that are induced during differentiation of CAD cells (Induced), shows reduced expression in ACSS2i-treated CAD neurons (inh.) compared to DMSO-treated control neurons (con.). Inhibitor-treated versus control, P&lt;2.2×10 −16 . ( FIG. 2F ) Nuclear acetyl-CoA levels are reduced in response to either knockdown of ACSS2 (shACSS2; mean Δ=−0.19±0.03, * * P=0.003) or application of the ACSS2 inhibitor (mean Δ=−0.25±0.05, * * P=0.006; n=3, mean+s.d.). ( FIG. 2G ) Western blot analysis of whole-cell lysates shows that lentiviral shRNA-mediated knockdown of ACSS2 lowers H3K9 and H3K27 acetylation (quantified in  FIG. 10A ). ( FIG. 2H ) Western blot analysis of immunoprecipitation eluates shows that CBP is co-immunoprecipitated with ACSS2 but not with control Ig. ( FIG. 2I ) Immunofluorescence in primary hippocampal neurons shows nuclear localization of ACSS2 (day 7 of in vitro differentiation culture, isolated from C57BL/6 embryos). Scale bar, 50 i m. ( FIG. 2J ) Western blots of lysates from primary hippocampal neurons (d7) treated for 24 hours with ACSS2i and probed with the indicated antibodies (quantified in  FIG. 10C ) show reduction of histone acetylation. 
         FIG. 3 , comprising  FIG. 3A  through  FIG. 3F , depicts results from example experiments demonstrating that ACSS2 ChIP-seq localization is linked to histone acetylation in vivo in mouse hippocampus. ( FIG. 3A ) ChIP-seq for ACSS2 and H3K9ac in mouse hippocampus. Track views show ACSS2 and H3K9ac for three neuronal genes involved in memory: Arc, Egr2 and Nr2f2 (chr15:74,496,025-74,506,488; chr10:66,991,018-67,006,804; and chr7:77,488,549-77,516,626, respectively). ( FIG. 3B ) In vivo hippocampal ACSS2 and H3K9ac peaks co-localize with the nearest gene TSS (&lt;1 kb from peak) among all RefSeq transcripts. ( FIG. 3C ) RNA-seq expression in dorsal hippocampus (dHPC) correlates with hippocampal ACSS2 binding and enrichment of H3K9 acetylation. ( FIG. 3D ) Expression profile of genes classified by their ACSS2 and H3K9ac enrichment state. ( FIG. 3E ) Overlap between ACSS2 targeted genes (hippocampus) and CBP and H3K27ac enrichment for entire set of peaks (ENCODE CBP and H3K27ac ChIP-seq in mouse forebrain and cortex). ( FIG. 3F ) Motif analysis at ACSS2 peaks from in vivo ChIP-seq in hippocampus showing top enrichment of NRF1, a neuronal transcription factor. 
         FIG. 4 , comprising  FIG. 4A  through  FIG. 4F , depicts results from example experiments demonstrating that ACSS2 knockdown in dorsal hippocampus impairs object location memory and upregulation of immediate early genes following training. ( FIG. 4A ) Stereotactic surgery was performed to deliver AAV9 knockdown vector into the dorsal hippocampus (AP, −2.0 mm; DV, −1.4 mm; ML, +1.5 mm from bregma); 4 weeks later, habituated mice were trained in object location memory (OLM; four 5-min training sessions in arena with three different objects). Twenty-four hours later the mice were given a retention test in which one object was moved to a novel location (n=10 per cohort). ( FIG. 4B ) Western blot analysis of hippocampal tissue removed from mice injected into the dorsal (d) or ventral (v) hippocampus with either eGFP control or ACSS2 knockdown vector shows specific reduction of ACSS2 in dorsal hippocampus. ( FIG. 4C ) ACSS2-knockdown mice are impaired in object location memory. eGFP control and shACSS2 AAV9 mice display no preference for any of three objects (O1-3) during the object location memory training session (TR). In the retention test 24 hours later, control mice show a preference for the novel object location (NL), whereas the knockdown mice display no such preference. *** P&lt;0.001; n=10, mean±s.d. ( FIG. 4D ) The spatial memory defect in ACSS2-knockdown mice manifests in a lowered discrimination index (% DI=(t NL−t FL)/(t NL+t FL)) compared to control mice (Δ DI=−29.5±11.4, * P=0.02; n=10, mean±s.d.). ( FIG. 4E ) Training-induced expression of a cohort of immediate early genes ( FIG. 12H ) is greatly attenuated in ACSS2-knockdown mice (n=4 mice per group, 2 replicates for each condition, P&lt;0.0001, paired t-test, mean±s.d.). ( FIG. 4F ) Model for function of ACSS2 as a chromatin-bound coactivator to provide acetyl-CoA locally to promote histone acetylation and activity-induced upregulation of immediate early genes. 
         FIG. 5 , comprising  FIG. 5A  through  FIG. 5G , depicts results from example experiments demonstrating that ACSS2 localizes to the nucleus of neurons. ( FIG. 5A ) Percentage of cells with nuclear staining in ACSS2 immunofluorescence experiments (undiff., undifferentiated CAD cells; diff., differentiated CAD neurons; hippocampal, primary hippocampal neurons day 7; a minimum of 50 cells were examined in three replicate immunofluorescence experiments; t-test undiff. vs diff. P&lt;0.0001, undiff. vs hippocampal P&lt;0.0001; error bars, s.e.m.). ( FIG. 5B ) Western blots of cytoplasmic (CE) and nuclear (NE) extracts from undifferentiated CAD cells and differentiated CAD neurons were probed with the indicated antibodies. ( FIG. 5C ,  FIG. 5D ) Immunofluorescence in primary cortical neurons isolated from C57BL/6 embryos, at day 7 ( FIG. 5C ) and day 14 ( FIG. 5D ) of in vitro differentiation culture. ACSS2 locates predominantly to nuclei in differentiated primary cortical neurons. All scale bars, 25 μm. ( FIG. 5E ) Immunofluorescence in primary hippocampal neurons isolated from C57BL/6 embryos at day 14 of in vitro differentiation culture. ACSS2 locates predominantly to nuclei in differentiated primary neurons. ( FIG. 5F ) Immunofluorescence in primary hippocampal neurons at day 7 shows that ACL is chiefly localized to the cytoplasm. ( FIG. 5G ) Neuronal differentiation markers decrease in ACSS2 knockdown cells. CAD cells were infected with lentiviral control (WT) or knockdown vector (shACSS2). Western blots of lysates from stably infected differentiated cells were probed with the indicated antibodies and quantified using ImageJ (n=3; error bars, s.e.m.). 
         FIG. 6 , comprising  FIG. 6A  through  FIG. 6O , depicts results from example experiments demonstrating that ACSS2 regulates neuronal gene expression. ( FIG. 6A ,  FIG. 6B ) Correlation plots of replicate RNA-seq in undifferentiated CAD cells ( FIG. 6A ) and differentiated CAD neurons ( FIG. 6B ) for scramble control. ( FIG. 6C ) Transcriptome analysis via RNA-seq, done in two highly correlated biological replicates, identified 894 genes that became upregulated in differentiated CAD neurons (red dots depict genes with &gt;1.6-fold increase). ( FIG. 6D ) Pathway analysis of the 894 upregulated genes (red dots in  FIG. 2A ) using StringDB. The protein-protein interaction graph depicts a network of binding partners that centers on key players in activity-dependent signaling and synaptic plasticity: Itpr1, Grin1, Nefh, Dynclh1 and Calm1. ( FIG. 2E ) Gene ontology enrichment analysis shows upregulation of neuronal pathways. Gene ontology analysis was used on the 894 genes that become upregulated in differentiated CAD neurons ( FIG. 6C ; identified by RNA-seq, fold-enrichment (FE) &gt;3.5, FDR&lt;0.005). ( FIG. 6F ) Genome browser view of Nudt from RNA-seq and ChIP-seq (H4K12ac, H4K5ac, and H3K9ac: mm10 chr5: 140,327,500-140,339,000). ( FIG. 6G ) Relative gene enrichment of H3K9ac, H4K5ac, and H4K12ac at genes that are upregulated during CAD neuron differentiation (&gt;1.6-fold, grey bars) versus all other genes (black bars). ( FIG. 6H ,  FIG. 6I ) Correlation plots of replicate RNA-seq in undifferentiated CAD cells for ACL knockdown ( FIG. 6H ), and ACSS2 knockdown ( FIG. 6I ). ( FIG. 6J ,  FIG. 6K ) Correlation plots of replicate RNA-seq in differentiated CAD neurons for ACL knockdown ( FIG. 6J ) and ACSS2 knockdown ( FIG. 6K ). ( FIG. 6L ) ACL knockdown has a much smaller effect on differentiation-linked upregulation of neuronal gene expression (compare to  FIG. 1D ). Scatter plot contrasts the fold-change FPKM of induced genes ( FIG. 6C ) between wild-type and ACL knockdown cells. Marginal distributions show histogram and kernel density estimation. Ordinary least squares linear regression is displayed with 95% confidence interval. ( FIG. 6M ) The corresponding quintiles of upregulated genes (red dots in  FIG. 6C ) with the greatest fold-change FPKM increase in wild-type cells. The ACL knockdown showed the same upward trend as the wild-type cells (red bars, compared to black bars in  FIG. 1F ), contrasting with the severe defect in ACSS2-knockdown cells (green bars; for each quintile, columns represent the mean induction value of genes and error bars represent s.e.m.). ( FIG. 6N ) Box plot of global mRNA transcript levels in undifferentiated and differentiated CAD neurons from RNA-seq in wild-type (scramble control knockdown; grey), ACSS2-knockdown (shACSS2 #25 knockdown; green), and ACL-knockdown (shACL #17 knockdown; red) cells. Genome-wide transcript levels are reduced in differentiated ACL-knockdown cells when compared to differentiated wild-type cells (error bars, s.e.m.), whereas the global reduction in differentiated ACSS2-knockdown cells is less significant when compared to differentiated wild-type cells (error bars, s.d.). ( FIG. 6O ) Genes sensitive to ACSS2 knockdown (top 20%) are also sensitive to ACSS2i treatment, which lowers their expression compared to all genes. 
         FIG. 7 , comprising  FIG. 7A  through  FIG. 7P , depicts results from example experiments demonstrating that ACSS2 is chromatin-bound in differentiated CAD neurons. ( FIG. 7A ) ChIP-seq in differentiated CAD neurons was performed in replicate with two different antibodies against ACSS2. Correlation plot displays relative enrichment over corresponding MACS peaks (default parameters with input as control, 1,598 peaks). ( FIG. 7B ) Correlation plot displays relative genome-wide ChIP-seq enrichment. ( FIG. 7C ) UCSC Genome Browser views of ChIP-seq tracks show that, upon CAD neuron differentiation, increases in H4K5, H4K12, and H3K9 acetylation over the Nudt1 locus co-occur with ACSS2 enrichment (chr5: 140,326,845-140,339,655). ( FIG. 7D ) UCSC Genome Browser view of indicated ChIP-seq tracks in undifferentiated CAD cells and differentiated CAD neurons over Tab2 locus (chr10: 7,875,000-8,004,000). ( FIG. 7E ) Gene ontology enrichment analysis of the genes most proximate to ACSS2 peaks demonstrates that neuron-specific genes are enriched. ( FIG. 7F ) Frequency of ACSS2 peaks (T antibody) located upstream of their target gene associated with histone acetylation. ( FIG. 7G ) Frequency of ACSS2 peaks (CS antibody) located upstream of their target gene associated with histone acetylation. ( FIG. 7H ) Table shows percent direct overlap of ACSS2 peaks with H3K9ac, H4K5ac, and H4K12ac broad MACS peaks. ( FIG. 7I ,  FIG. 7J ,  FIG. 7K ) Decile plots depict enrichment of H3K9ac ( FIG. 7I ), H4K5ac ( FIG. 7J ), and H4K12ac ( FIG. 7K ) over ranked deciles of ACSS2 peak enrichment (zeroes removed). ( FIG. 7L ,  FIG. 7M ,  FIG. 7N ) Differentiation-induced co-enrichment of ACSS2 and acetyl broad peaks (MACS). Peak enrichment correlation indicated for H3K9ac ( FIG. 7L ), H4K5ac ( FIG. 7M ), and H4K12ac ( FIG. 7N ). ( FIG. 7O ) Discovered de novo motifs for transcription factor binding sites predicted by HOMER from all ACSS2 ChIP-seq peaks called by MACS in differentiated CAD neurons. ( FIG. 7P ) ChIP-seq enrichment of differentiation-induced genes as a group shows correlation with histone acetylation in differentiated CAD neurons. 
         FIG. 8 , comprising  FIG. 8A  and  FIG. 8B , depicts results from example experiments demonstrating that ACSS2 enrichment co-occurs with histone acetylation at neuronal genes in differentiating CAD neurons. ( FIG. 8A ) UCSC Genome Browser views of ChIP-seq tracks demonstrate that increases in H4K5, H4K12, and H3K9 acetylation co-occur with ACSS2 enrichment over the Idua (α-1-iduronidase) locus upon CAD neuron differentiation (chr5: 108,649,457-108,687,261). ( FIG. 8B ) At the Slc19A1 (solute carrier family 19 member 1) gene, elevated histone H4K5, H4K12, and H3K9 acetylation levels correspond with increasing ACSS2 enrichment in differentiated CAD neurons (chr10: 76,761,141-77,170,455). 
         FIG. 9 , comprising  FIG. 9A  through  FIG. 9I , depicts results from example experiments demonstrating that genic ACSS2 enrichment upon CAD neuronal differentiation corresponds to increased histone acetylation. ( FIG. 9A ,  FIG. 9B ,  FIG. 9C ,  FIG. 9D ) Metagene enrichment analysis shows ChIP occupancy for ACSS2 ( FIG. 9A ), H3K9ac ( FIG. 9B ), H4K5ac ( FIG. 9C ) and H4K12ac ( FIG. 9D ) across the top 5% of genes enriched for ACSS2 in differentiated CAD neurons (Top 5% DE; red). The bottom 80% of genes (Bot 80% DE) is shown in blue, and the average signal across all genes (All genes DE) is shown in green. ( FIG. 9E ,  FIG. 9F ,  FIG. 9G ,  FIG. 9H ) Meta-gene enrichment analysis shows ChIP occupancy for ACSS2 ( FIG. 9E ), H3K9ac ( FIG. 9F ), H4K5ac ( FIG. 9G ) and H4K12ac ( FIG. 9H ) at the top 5% of genes that become dynamically bound by ACSS2 upon neuronal differentiation (Top 5% DE; red). The bottom 80% of genes (Bot 80% DE) is shown in blue, and the average signal across all genes (All genes DE) is shown in green. ( FIG. 9I ) Multiple linear regression analysis was implemented to model the interaction between genic ACSS2 enrichment and wild-type gene expression changes, and to visualize the interaction between differentiation-linked gene expression changes and ACSS2 recruitment to chromatin. The contour plot of this fitted regression model displays high levels of ACSS2 enrichment in red and low levels in blue, and is overlaid with the scatter plot of the independent gene expression variables. The visualized model demonstrates that high ACSS2 enrichment corresponds to increased gene expression in differentiated CAD neurons. 
         FIG. 10 , comprising  FIG. 10A  through  FIG. 10C , depicts results from example experiments demonstrating that ACSS2 functions in neuronal histone acetylation. ( FIG. 10A ) Western blot analysis of whole-cell lysates shows that lentiviral shRNA-mediated knockdown of ACSS2 lowers H3K9 and H3K27 acetylation (compare to  FIG. 2G ), quantified using ImageJ (n=3, error bars show s.e.m.). ( FIG. 10B ) Western blot analysis of eluates and supernatants of IgG control and ACSS2 co-immunoprecipitation experiments indicates that ACSS2 binds to acetylated chromatin. ( FIG. 10C ) Western blots of lysates from primary hippocampal neurons (day 7) treated for 24 hours with the ACSS2i, probed with the indicated antibodies (compare to  FIG. 2J ), and quantified using ImageJ (n=3, error bars show s.e.m.). 
         FIG. 11 , comprising  FIG. 11A  through  FIG. 11C , depicts results from example experiments demonstrating that ACSS2 chromatin association and H3K9ac in dorsal hippocampus corresponds to H3K27ac and CBP enrichment in neuronal tissue. ( FIG. 11A ) Genome-wide compartment analysis of in vivo hippocampal ChIP-seq of H3K9ac and mouse forebrain H3K9ac ChIP-seq from ENCODE, showing a similar peak distribution genome-wide: although they originate in different brain regions, the in vivo H3K9ac ChIP data are in strong agreement (Spearman R=0.67). ( FIG. 11B ) Overlap of RefSeq transcripts targeted by the indicated enzyme or modification (peaks for CBP (GSM1629373) and H3K27ac (GSM1629397) in mouse cortical neurons were called using MACS2 (narrow peaks, FDR 0.1%) with an input sonication efficiency control (GSM1629381); peaks were associated to the nearest TSS among all RefSeq transcripts). ( FIG. 11C ) Gene Ontology enrichment analysis performed on common CBP-ACSS2 targets, indicating that these enzymes co-target genes that modulate synapse biology and synaptic membrane potential. 
         FIG. 12 , comprising  FIG. 12A  through  FIG. 12H , depicts results from example experiments demonstrating that attenuation of ACSS2 expression in the dorsal hippocampus impairs object location memory. ( FIG. 12A ) ACSS2 RNA in situ hybridization on ACSS2 in sagittal section of hippocampal region CA1 (left, reference atlas adapted from Allen Mouse Brain Atlas12; right, in situ hybridization; HPC, hippocampus proper). ( FIG. 12B ) Weight of eGFPAAV9 control and shACSS2-AAV9 knockdown mice before intracranial surgery, and following recovery before object location memory (OLM) training (NS, n=10 per group, error bars show s.d.). ( FIG. 12C ,  FIG. 12D ) ACSS2 knockdown mice showed no defect in locomotion or thigmotaxis (tendency to remain close to vertical surfaces in an open field, a measure of anxiety), as quantified over 5 min in the open field test; ( FIG. 12C ) shows example heat map of tracking data (NS, n=10 per group, error bars show s.d.). ( FIG. 12E ) Exploration times by eGFP-AAV9 control and shACSS2-AAV9 knockdown mice recorded for the three objects (O1-3) during the first OLM training session (TR) and the 24-h retention test (NL, object in novel location; FL, objects in former location). ( FIG. 12F ) Compared to the control eGFPAVV9 mice, ACSS2-knockdown mice showed no defect in contextual fear memory. Freezing in chamber on day of contextual fear conditioning was recorded and quantified pre-shock (FC Training; NS, n=10 per cohort, error bars show s.d.). Fear memory was measured as the freezing response after re-exposure to the context 1 day after contextual fear conditioning (aversive stimulus: 1.5 mA electrical shock; NS, n=10 per cohort, error bars show s.d.). ( FIG. 12G ) RNA-seq was performed on the dorsal hippocampus of eGFP control and shACSS2-knockdown animals. Global transcript levels were not affected by ACSS2 knockdown (dHPC, dorsal hippocampus; four animals per group, two replicates for each condition; NS, paired t-test, error bars show s.d.). ( FIG. 12H ) Baseline expression of immediate-early genes in untrained animals was unaltered in ACSS2-knockdown mice. RNA-seq was performed on the dorsal hippocampus of eGFP control and shACSS2-knockdown mice (r=0.82, P&lt;0.0001; HCC, homecage circadian control). 
         FIG. 13 , comprising  FIG. 13A  through  FIG. 13F , depicts results from example experiments demonstrating that ACSS2 regulates retrieval-induced upregulation of immediate-early genes in vivo. ( FIG. 13A ) Genome-wide RNA-seq was performed on the dorsal hippocampus of eGFP control and shACSS2-knockdown mice. The analysis was focused on the set of previously identified and validated genes that become upregulated during the sensitive period following memory retrieval. The baseline expression of immediate early genes in untrained animals was not changed in shACSS2-AAV9 mice when compared to eGFP-AAV9 control mice (CC, circadian control). ( FIG. 13B ) During the sensitive period following contextual memory retrieval (RT, 30 min post-exposure to conditioning chamber 24 hours after fear conditioning), immediate early genes were upregulated in the dorsal hippocampus of control injected mice. By contrast, the dynamic retrieval-induced expression of these early response genes is absent in ACSS2-knockdown mice (P=0.001, paired t-test). ( FIG. 13C ) Induction defect of immediate early genes in shACSS2-AAV9 injected animals (RT/CC). ( FIG. 13D ) The baseline expression of genes that were downregulated after contextual memory retrieval is not altered in ACSS2-knockdown mice. ( FIG. 13E ) Downregulation of retrieval-responsive genes occurs in both eGFP control and ACSS2-knockdown mice, except for Cldn5. ( FIG. 13F ) Retrieval-induced downregulation of retrieval-responsive genes in the dorsal hippocampus in eGFP control versus shACSS2-knockdown mice (RT/CC). 
         FIG. 14  depicts the original gel blots of the western blots depicted in  FIGS. 1C, 2G, 1E and 5G . Boxes depict the cropped area shown in  FIGS. 1C, 2G, 1E and 5G . 
         FIG. 15  depicts the original gel blots of the western blots depicted in  FIGS. 2H and 2J . Boxes depict the cropped area shown in  FIGS. 2H and 2J . 
         FIG. 16  depicts the original gel blots of the western blots depicted in  FIGS. 4B, 5B and 10B . Boxes depict the cropped area shown in  FIGS. 4B, 5B and 10B . 
         FIG. 17  depicts the physiological sources of acetate. 
         FIG. 18  depicts graphs measuring the histone acetylation in the cortex, hippocampus, and liver of mice after intraperitoneal injection of EtOH- 13 C 2 . 
         FIG. 19  depicts graphs measuring the histone acetylation in the hippocampus of ACSS2-knockdown mice. 
         FIG. 20  depicts graphs demonstrating the difference in histone acetylation in the dorsal HPC, ventral HPC and muscle of ACSS2-knockdown mice versus wild type mice. 
         FIG. 21 , comprising  FIG. 21A  through  FIG. 21E , depicts that alcohol metabolites feed histone acetylation in the brain.  FIG. 21A  depicts an experimental outline of in vivo EtOH-d 6  mass spectrometry.  FIG. 21B  depicts experimental results demonstrating metabolized heavy EtOH-d 6  is incorporated into histone acetylation in hippocampus. The Arachne plot axis represents the % of the third isotope for the acetylated peptide, corresponding to the D 3  labeled form; the natural relative abundance of that isotope is apparent in the ‘none’ and ‘saline 1h’ treatment groups.  FIG. 21C  depicts experimental results demonstrating label incorporation into cortical histone acetylation shows a similar pattern to the hippocampus.  FIG. 21D  depicts experimental results demonstrating label incorporation into histone acetylation occurs earlier in the liver, the principal site of alcohol metabolism.  FIG. 21E  depicts experimental results demonstrating histone acetylation is relatively independent of liver alcohol metabolism in skeletal muscle, a tissue with low expression of ACSS2. 
         FIG. 22 , comprising  FIG. 22A  through  FIG. 22D , depicts histone acetylation of wild-type mice.  FIGS. 22A-22C  depict mass spectra showing the relative abundance of deuterated histone H4-triacetyl peptide (aa 4-17) in dorsal hippocampus of wild-type mice.  FIG. 22A  depicts the mass spectrum at baseline.  FIG. 22B  depicts the mass spectrum at 30 minutes following d6-EtOH injection.  FIG. 22C  depicts the mass spectrum at 4 hours following d6-EtOH injection.  FIG. 22D  depicts experimental results demonstrating histone acetylation is relatively independent of liver alcohol metabolism in skeletal muscle. Relative abundance of deuterated histone acetylation in skeletal muscle tissue at 30 minutes and 4 hours in wild type (WT) mice, and 30 minutes in hippocampal ACSS2 KD mice (compare to  FIG. 21E ). 
         FIG. 23 , comprising  FIG. 23A  through  FIG. 23E , depicts mass spectrometry analysis of EtOH-d 6  in a dorsal hippocampus (dHPC) ACSS2 knockdown (KD).  FIG. 23A  depicts experimental results demonstrating knockdown of ACSS2 expression in dorsal hippocampus prevents incorporation of the heavy label into histone acetylation.  FIG. 23B  depicts experimental results demonstrating that, in the same animal, incorporation of the heavy label in the ventral hippocampus (where ACSS2 levels are normal) is not changed when compared to control mice.  FIG. 23C  depicts experimental results demonstrating heavy acetate introduced via intraperitoneal injection readily labels histone acetylation in the dorsal hippocampus.  FIG. 23D  depicts experimental results demonstrating heavy acetate introduced via intraperitoneal injection readily labels histone acetylation in the cortex.  FIG. 23E  depicts experimental results demonstrating acetate from hepatic alcohol breakdown is activated by neuronal ACSS2 in the brain and readily induces gene-regulatory histone acetylation. 
         FIG. 24 , comprising  FIG. 24A  through  FIG. 24E , depicts ACSS2 mediated acetate-induced transcription in primary hippocampal neurons.  FIG. 24A  and  FIG. 24B  depict RNA-seq in primary hippocampal neurons isolated from C57/Bl6 mouse embryos and treated with acetate (10 mM) in the presence or absence of a small molecular inhibitor of ACSS2 (ACSS2i).  FIG. 24A  depicts a heatmap showing 7,600 genes differentially expressed upon acetate treatment, and a third column showing the behavior of those genes under in the presence of the ACSS2 inhibitor. 2107 of the 3613 acetate-induced genes fail to be upregulated in the presence of ACSS2i (N=4 per group).  FIG. 24B  depicts experimental results demonstrating acetate-induced genes were not regulated by ACSS2i treatment in the absence of acetate.  FIG. 24C  depicts acetate-induced genes in primary hippocampal neurons in blue; shown below the Gene Ontology (GO) term analysis of ACSS2i sensitive genes (non-overlapping with yellow, acetate+ACSS2i).  FIG. 24D  depicts GO term analysis of genes that are both sensitive to acetate and directly bound by ACSS2 (from ACSS2 ChIP-seq).  FIG. 24E  depicts HOMER unsupervised de novo motif analysis of ACSS2 hippocampal binding sites targeting acetate-sensitive genes. 
         FIG. 25 , comprising  FIG. 25A  through  FIG. 25D , depicts genes regulated by acetate.  FIG. 25A  depicts RNAseq showing differentially regulated genes in primary hippocampal neurons treated with 10 mM acetate.  FIG. 25B  depicts gene ontology (GO) analysis of significantly upregulated (red) and significantly downregulated (blue) genes.  FIG. 25C  experimental results demonstrating 81 out of 214 genes upregulated in the hippocampus of ethanol-injected mice are also upregulated by acetate in primary hippocampal neurons in vitro (p=6.60e-17).  FIG. 25D  depicts experimental results demonstrating the cumulative number of ACSS2 peaks near the transcription start site (TSS) of acetylated ACSS2i sensitive genes, indicating that the majority ACSS2 binding events occurs over or proximal to the gene promoter. 
         FIG. 26 , comprising  FIG. 26A  and  FIG. 26B , depicts that alcohol metabolites feed histone acetylation in the fetal brain.  FIG. 26A  depicts experimental results demonstrating metabolized heavy d6-EtOH is incorporated into histone acetylation in the maternal brain.  FIG. 26B  depicts heavy label incorporation into histone acetylation in the fetal brain. Data represent two pools of four embryos from maternal d6-EtOH injection. The Arachne plot axes represent the percentage of the third isotope of the acetylated peptide, corresponding to the D3 labeled form. 
         FIG. 27  depicts experimental results demonstrating the peptide H4 aa 4-17 with 3 acetyls (hippocampus). 
         FIG. 28  depicts experimental results of SILAC-mass spec experiments. 
         FIG. 29  depicts assay design to determine efficacy to reduce catalytic ACSS2 activity and histone H3 lysine 9 acetylation in vitro—Ntera2 cells were maintained in DMEM (Gibco) with 10% FBS and GlutaMAX (Gibco). Cells were treated for 24 hours with 5 mM sodium acetate in the absence of glucose and compound ADG-204, ADG-205, ADG-206, or vehicle (DMSO). Cells were lysed in RIPA buffer containing 50 mM Tris pH 8.0, 0.5 mM EDTA, 150 mM NaCl, 1% NP40, 1% SDS, supplemented with protease inhibitor cocktail (Life Technologies, number 78446) and 10 mM sodium butyrate. Protein concentration was determined by BCA protein assay (Life Technologies, number 23227), and equal amounts of protein were directly loaded onto polyacrylamide gels. Proteins were separated on 4-12% Bis-Tris polyacrylamide gels (NuPAGE). After transfer to nitrocellulose membrane, 3% BSA in TBS supplemented with 0.1% Tween 20 (TBST) was used to block the membrane at room temperature for 1 h. Primary antibodies were diluted in TBST and incubated at 4° C. overnight. The antibodies used were anti-H3 (Abcam ab1791), anti-H3K9ac (Abcam ab4441), anti-GAPDH (Fitzgerald Industries 10R-G109A). The membrane was washed three times with TBST, each for 10 min, followed by incubation with HRP-conjugated secondary antibodies at room temperature for 1 h, in TBST. The membrane was washed again three times and imaged with a Fujifilm LAS-4000 imager. 
         FIG. 30  depicts the chemical structure and activity of ADG-204. 
         FIG. 31  depicts the chemical structure and activity of ADG-205. 
         FIG. 32  depicts the chemical structure and activity of ADG-206. 
     
    
    
     DETAILED DESCRIPTION 
     The present invention relates to compositions and methods for treating neurological and cognitive diseases and disorders. In some embodiments, the invention provides compositions and methods for treating memory-related diseases and disorders. In various embodiments, the compositions and methods of the invention are useful in treating anxiety diseases and disorders such as phobias, panic disorders, psychosocial stress (e.g. as seen in disaster, catastrophe or violence victims), obsessive-compulsive disorder, generalized anxiety disorder and post-traumatic stress disorder (PTSD). In some embodiments, the compositions and methods of the invention are useful for regulating long term memory storage or consolidation. 
     The present invention also relates to compositions and methods for treating addiction and/or disease or disorders related to addiction. In various embodiments, the compositions and methods of the invention are useful for preventing or treating acute alcohol induced memory deficit and chronic alcohol induced memory deficit. 
     In some embodiments, the methods of the present invention comprise modulating chromatin acetylation. In one embodiment, the methods of the invention decrease chromatin acetylation. In one embodiment, the chromatin is neuronal chromatin. In one embodiment, the method comprises administering to a subject an effective amount of a composition comprising an inhibitor of ACSS2. 
     In certain instances, the compositions and methods described herein relate to inhibiting acetate-dependent acetyl-CoA synthetase 2 (ACSS2). In one embodiment, the composition of the present invention comprises an inhibitor of ACSS2. In one embodiment, the inhibitor of ACSS22 inhibits the expression, activity, or both, of ACSS2. 
     Definitions 
     Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. 
     Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and nucleic acid chemistry and hybridization are those well-known and commonly employed in the art. 
     Standard techniques are used for nucleic acid and peptide synthesis. The techniques and procedures are generally performed according to conventional methods in the art and various general references (e.g., Sambrook and Russell, 2012, Molecular Cloning, A Laboratory Approach, Cold Spring Harbor Press, Cold Spring Harbor, N.Y., and Ausubel et al., 2002, Current Protocols in Molecular Biology, John Wiley &amp; Sons, NY), which are provided throughout this document. 
     As used herein, each of the following terms has the meaning associated with it in this section. 
     The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. “About” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. 
     The term “abnormal” when used in the context of organisms, tissues, cells or components thereof, refers to those organisms, tissues, cells or components thereof that differ in at least one observable or detectable characteristic (e.g., age, treatment, time of day, etc.) from those organisms, tissues, cells or components thereof that display the “normal” (expected) respective characteristic. Characteristics which are normal or expected for one cell or tissue type, might be abnormal for a different cell or tissue type. 
     “Antisense” refers particularly to the nucleic acid sequence of the non-coding strand of a double stranded DNA molecule encoding a protein, or to a sequence which is substantially homologous to the non-coding strand. As defined herein, an antisense sequence is complementary to the sequence of a double stranded DNA molecule encoding a protein. It is not necessary that the antisense sequence be complementary solely to the coding portion of the coding strand of the DNA molecule. The antisense sequence may be complementary to regulatory sequences specified on the coding strand of a DNA molecule encoding a protein, which regulatory sequences control expression of the coding sequences. 
     A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal&#39;s health continues to deteriorate. 
     In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal&#39;s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal&#39;s state of health. 
     A disease or disorder is “alleviated” if the severity of a sign or symptom of the disease or disorder, the frequency with which such a sign or symptom is experienced by a patient, or both, is reduced. 
     An “effective amount” or “therapeutically effective amount” of a compound is that amount of a compound which is sufficient to provide a beneficial effect to the subject to which the compound is administered. 
     “Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA. 
     The terms “patient,” “subject,” “individual,” and the like are used interchangeably herein, and refer to any animal, or cells thereof whether in vitro or in vivo, amenable to the methods described herein. In certain non-limiting embodiments, the patient, subject or individual is a human. 
     A “therapeutic” treatment is a treatment administered to a subject who exhibits signs or symptoms of a disease or disorder, for the purpose of diminishing or eliminating those signs or symptoms. 
     As used herein, “treating a disease or disorder” means reducing the severity and/or frequency with which a sign or symptom of the disease or disorder is experienced by a patient. 
     By the term “specifically binds,” as used herein with respect to an antibody, is meant an antibody which recognizes a specific antigen, but does not substantially recognize or bind other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. But, such cross-species reactivity does not itself alter the classification of an antibody as specific. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, such cross reactivity does not itself alter the classification of an antibody as specific. 
     In some instances, the terms “specific binding” or “specifically binding,” can be used in reference to the interaction of an antibody, a protein, or a peptide with a second chemical species, to mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody. 
     A “coding region” of a gene consists of the nucleotide residues of the coding strand of the gene and the nucleotides of the non-coding strand of the gene which are homologous with or complementary to, respectively, the coding region of an mRNA molecule which is produced by transcription of the gene. 
     A “coding region” of a mRNA molecule also consists of the nucleotide residues of the mRNA molecule which are matched with an anti-codon region of a transfer RNA molecule during translation of the mRNA molecule or which encode a stop codon. The coding region may thus include nucleotide residues comprising codons for amino acid residues which are not present in the mature protein encoded by the mRNA molecule (e.g., amino acid residues in a protein export signal sequence). 
     “Complementary” as used herein to refer to a nucleic acid, refers to the broad concept of sequence complementarity between regions of two nucleic acid strands or between two regions of the same nucleic acid strand. It is known that an adenine residue of a first nucleic acid region is capable of forming specific hydrogen bonds (“base pairing”) with a residue of a second nucleic acid region which is antiparallel to the first region if the residue is thymine or uracil. Similarly, it is known that a cytosine residue of a first nucleic acid strand is capable of base pairing with a residue of a second nucleic acid strand which is antiparallel to the first strand if the residue is guanine. A first region of a nucleic acid is complementary to a second region of the same or a different nucleic acid if, when the two regions are arranged in an antiparallel fashion, at least one nucleotide residue of the first region is capable of base pairing with a residue of the second region. In one embodiment, the first region comprises a first portion and the second region comprise a second portion, whereby, when the first and second portions are arranged in an antiparallel fashion, at least about 50% of the nucleotide residues of the first portion are capable of base pairing with nucleotide residues in the second portion. In one embodiment, at least about 75%, at least about 90%, or at least about 95% of the nucleotide residues of the first portion are capable of base pairing with nucleotide residues in the second portion. In one embodiment, all nucleotide residues of the first portion are capable of base pairing with nucleotide residues in the second portion. 
     The term “DNA” as used herein is defined as deoxyribonucleic acid. 
     The term “expression” as used herein is defined as the transcription and/or translation of a particular nucleotide sequence driven by its promoter. 
     The term “expression vector” as used herein refers to a vector containing a nucleic acid sequence coding for at least part of a gene product capable of being transcribed. In some cases, RNA molecules are then translated into a protein, polypeptide, or peptide. In other cases, these sequences are not translated, for example, in the production of antisense molecules, siRNA, ribozymes, and the like. Expression vectors can contain a variety of control sequences, which refer to nucleic acid sequences necessary for the transcription and possibly translation of an operatively linked coding sequence in a particular host organism. In addition to control sequences that govern transcription and translation, vectors and expression vectors may contain nucleic acid sequences that serve other functions as well. 
     The term “fusion polypeptide” refers to a chimeric protein containing a protein of interest (e.g., luciferase) joined to a heterologous sequence (e.g., a non-luciferase amino acid or protein). 
     The term “homology” refers to a degree of complementarity. There may be partial homology or complete homology (i.e., identity). Homology is often measured using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group. University of Wisconsin Biotechnology Center. 1710 University Avenue. Madison, Wis. 53705). Such software matches similar sequences by assigning degrees of homology to various substitutions, deletions, insertions, and other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. 
     “Isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in its normal context in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural context is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell. 
     The term “isolated” when used in relation to a nucleic acid, as in “isolated oligonucleotide” or “isolated polynucleotide” refers to a nucleic acid sequence that is identified and separated from at least one contaminant with which it is ordinarily associated in its source. Thus, an isolated nucleic acid is present in a form or setting that is different from that in which it is found in nature. In contrast, non-isolated nucleic acids (e.g., DNA and RNA) are found in the state they exist in nature. For example, a given DNA sequence (e.g., a gene) is found on the host cell chromosome in proximity to neighboring genes; RNA sequences (e.g., a specific mRNA sequence encoding a specific protein), are found in the cell as a mixture with numerous other mRNAs that encode a multitude of proteins. However, isolated nucleic acid includes, by way of example, such nucleic acid in cells ordinarily expressing that nucleic acid where the nucleic acid is in a chromosomal location different from that of natural cells, or is otherwise flanked by a different nucleic acid sequence than that found in nature. The isolated nucleic acid or oligonucleotide may be present in single-stranded or double-stranded form. When an isolated nucleic acid or oligonucleotide is to be utilized to express a protein, the oligonucleotide contains at a minimum, the sense or coding strand (i.e., the oligonucleotide may be single-stranded), but may contain both the sense and anti-sense strands (i.e., the oligonucleotide may be double-stranded). 
     The term “isolated” when used in relation to a polypeptide, as in “isolated protein” or “isolated polypeptide” refers to a polypeptide that is identified and separated from at least one contaminant with which it is ordinarily associated in its source. Thus, an isolated polypeptide is present in a form or setting that is different from that in which it is found in nature. In contrast, non-isolated polypeptides (e.g., proteins and enzymes) are found in the state they exist in nature. 
     By “nucleic acid” is meant any nucleic acid, whether composed of deoxyribonucleosides or ribonucleosides, and whether composed of phosphodiester linkages or modified linkages such as phosphotriester, phosphoramidate, siloxane, carbonate, carboxymethylester, acetamidate, carbamate, thioether, bridged phosphoramidate, bridged methylene phosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, bridged phosphorothioate or sulfone linkages, and combinations of such linkages. The term nucleic acid also specifically includes nucleic acids composed of bases other than the five biologically occurring bases (adenine, guanine, thymine, cytosine and uracil). The term “nucleic acid” typically refers to large polynucleotides. 
     Conventional notation is used herein to describe polynucleotide sequences: the left-hand end of a single-stranded polynucleotide sequence is the 5′-end; the left-hand direction of a double-stranded polynucleotide sequence is referred to as the 5′-direction. 
     The direction of 5′ to 3′ addition of nucleotides to nascent RNA transcripts is referred to as the transcription direction. The DNA strand having the same sequence as an mRNA is referred to as the “coding strand”; sequences on the DNA strand which are located 5′ to a reference point on the DNA are referred to as “upstream sequences”; sequences on the DNA strand which are 3′ to a reference point on the DNA are referred to as “downstream sequences.” 
     By “expression cassette” is meant a nucleic acid molecule comprising a coding sequence operably linked to promoter/regulatory sequences necessary for transcription and, optionally, translation of the coding sequence. 
     The term “operably linked” as used herein refer to the linkage of nucleic acid sequences in such a manner that a nucleic acid molecule capable of directing the transcription of a given gene and/or the synthesis of a desired protein molecule is produced. The term also refers to the linkage of sequences encoding amino acids in such a manner that a functional (e.g., enzymatically active, capable of binding to a binding partner, capable of inhibiting, etc.) protein or polypeptide is produced. 
     As used herein, the term “promoter/regulatory sequence” means a nucleic acid sequence which is required for expression of a gene product operably linked to the promoter/regulator sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter/regulatory sequence may, for example, be one which expresses the gene product in a n inducible manner. 
     An “inducible” promoter is a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced substantially only when an inducer which corresponds to the promoter is present. 
     A “constitutive” promoter is a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell. 
     The term “polynucleotide” as used herein is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Thus, nucleic acids and polynucleotides as used herein are interchangeable. One skilled in the art has the general knowledge that nucleic acids are polynucleotides, which can be hydrolyzed into the monomeric “nucleotides.” The monomeric nucleotides can be hydrolyzed into nucleosides. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any means available in the art, including, without limitation, recombinant means, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using ordinary cloning technology and PCR, and the like, and by synthetic means. 
     In the context of the present invention, the following abbreviations for the commonly occurring nucleic acid bases are used. “A” refers to adenosine, “C” refers to cytosine, “G” refers to guanosine, “T” refers to thymidine, and “U” refers to uridine. 
     As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein&#39;s or peptide&#39;s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof. 
     As used herein, a “peptidomimetic” is a compound containing non-peptidic structural elements that is capable of mimicking the biological action of a parent peptide. A peptidomimetic may or may not comprise peptide bonds. 
     The term “RNA” as used herein is defined as ribonucleic acid. 
     “Recombinant polynucleotide” refers to a polynucleotide having sequences that are not naturally joined together. An amplified or assembled recombinant polynucleotide may be included in a suitable vector, and the vector can be used to transform a suitable host cell. 
     A recombinant polynucleotide may serve a non-coding function (e.g., promoter, origin of replication, ribosome-binding site, etc.) as well. 
     The term “recombinant polypeptide” as used herein is defined as a polypeptide produced by using recombinant DNA methods. 
     As used herein, “conjugated” refers to covalent attachment of one molecule to a second molecule. 
     As used herein, the term “transdominant negative mutant gene” refers to a gene encoding a polypeptide or protein product that prevents other copies of the same gene or gene product, which have not been mutated (i.e., which have the wild-type sequence) from functioning properly (e.g., by inhibiting wild type protein function). The product of a transdominant negative mutant gene is referred to herein as “dominant negative” or “DN” (e.g., a dominant negative protein, or a DN protein). 
     The phrase “inhibit,” as used herein, means to reduce a molecule, a reaction, an interaction, a gene, an mRNA, and/or a protein&#39;s expression, stability, function or activity by a measurable amount or to prevent entirely. Inhibitors are compounds that, e.g., bind to, partially or totally block stimulation, decrease, prevent, delay activation, inactivate, desensitize, or down regulate a protein, a gene, and an mRNA stability, expression, function and activity, e.g., antagonists. 
     “Variant” as the term is used herein, is a nucleic acid sequence or a peptide sequence that differs in sequence from a reference nucleic acid sequence or peptide sequence respectively, but retains essential biological properties of the reference molecule. Changes in the sequence of a nucleic acid variant may not alter the amino acid sequence of a peptide encoded by the reference nucleic acid, or may result in amino acid substitutions, additions, deletions, fusions and truncations. Changes in the sequence of peptide variants are typically limited or conservative, so that the sequences of the reference peptide and the variant are closely similar overall and, in many regions, identical. A variant and reference peptide can differ in amino acid sequence by one or more substitutions, additions, deletions in any combination. A variant of a nucleic acid or peptide can be a naturally occurring such as an allelic variant, or can be a variant that is not known to occur naturally. Non-naturally occurring variants of nucleic acids and peptides may be made by mutagenesis techniques or by direct synthesis. 
     A “vector” is a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to include non-plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, and the like. Examples of viral vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, and the like. 
     As used herein, the term “pharmaceutical composition” refers to a mixture of at least one compound useful within the invention with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary and topical administration. 
     As used herein, the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing an undesirable biological effect or interacting in a deleterious manner with any of the components of the composition in which it is contained. 
     As used herein, the language “pharmaceutically acceptable salt” refers to a salt of the administered compound prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids, organic acids, solvates, hydrates, or clathrates thereof. Examples of such inorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, acetic, hexafluorophosphoric, citric, gluconic, benzoic, propionic, butyric, sulfosalicylic, maleic, lauric, malic, fumaric, succinic, tartaric, amsonic, pamoic, p-tolunenesulfonic, and mesylic. Appropriate organic acids may be selected, for example, from aliphatic, aromatic, carboxylic and sulfonic classes of organic acids, examples of which are formic, acetic, propionic, succinic, camphorsulfonic, citric, fumaric, gluconic, isethionic, lactic, malic, mucic, tartaric, para-toluenesulfonic, glycolic, glucuronic, maleic, furoic, glutamic, benzoic, anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, pantothenic, benzenesulfonic (besylate), stearic, sulfanilic, alginic, galacturonic, and the like. Furthermore, pharmaceutically acceptable salts include, by way of non-limiting example, alkaline earth metal salts (e.g., calcium or magnesium), alkali metal salts (e.g., sodium-dependent or potassium), and ammonium salts. 
     As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer&#39;s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound useful within the invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the invention are known in the art and described, for example in Remington&#39;s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, Pa.), which is incorporated herein by reference. 
     As used herein, the term “potency” refers to the dose needed to produce half the maximal response (ED 50 ). 
     As used herein, the term “efficacy” refers to the maximal effect (E max ) achieved within an assay. 
     As used herein, the term “alkyl,” by itself or as part of another substituent means, unless otherwise stated, a straight or branched chain hydrocarbon having the number of carbon atoms designated (i.e. C1-6 means one to six carbon atoms) and including straight, branched chain, or cyclic substituent groups. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. 
     As used herein, the term “substituted alkyl” means alkyl as defined above, substituted by one, two or three substituents selected from the group consisting of halogen, —OH, alkoxy, —NH 2 , amino, azido, —N(CH 3 ) 2 , —C(═O)OH, trifluoromethyl, —C≡N, —C(═O)O(C 1 -C 4 )alkyl, —C(═O)NH 2 , —SO 2 NH 2 , —C(═NH)NH 2 , and —NO 2 . Examples of substituted alkyls include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl and 3-chloropropyl. 
     As used herein, the term “heteroalkyl” by itself or in combination with another term means, unless otherwise stated, a stable straight or branched chain alkyl group consisting of the stated number of carbon atoms and one or two heteroatoms selected from the group consisting of 0, N, and S, and wherein the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatom(s) may be placed at any position of the heteroalkyl group, including between the rest of the heteroalkyl group and the fragment to which it is attached, as well as attached to the most distal carbon atom in the heteroalkyl group. Examples include: —O—CH 2 —CH 2 —CH 3 , —CH 2 —CH 2 —CH 2 —OH, —CH 2 —CH 2 —NH—CH 3 , —CH 2 —S—CH 2 —C H 3 , and —CH 2 CH 2 —S(═O)—CH 3 . Up to two heteroatoms may be consecutive, such as, for example, —CH 2 —NH—OCH 3 , or —CH 2 —CH 2 —S—S—CH 3    
     As used herein, the term “alkoxy” employed alone or in combination with other terms means, unless otherwise stated, an alkyl group having the designated number of carbon atoms, as defined above, connected to the rest of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs and isomers. 
     As used herein, the term “halo” or “halogen” alone or as part of another substituent means, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. 
     As used herein, the term “cycloalkyl” refers to a mono cyclic or polycyclic non-aromatic radical, wherein each of the atoms forming the ring (i.e. skeletal atoms) is a carbon atom. In one embodiment, the cycloalkyl group is saturated or partially unsaturated. In another embodiment, the cycloalkyl group is fused with an aromatic ring. Cycloalkyl groups include groups having from 3 to 10 ring atoms. Illustrative examples of cycloalkyl groups include, but are not limited to, the following moieties: 
     
       
         
         
             
             
         
       
     
     Monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Dicyclic cycloalkyls include, but are not limited to, tetrahydronaphthyl, indanyl, and tetrahydropentalene. Polycyclic cycloalkyls include adamantine and norbornane. The term cycloalkyl includes “unsaturated nonaromatic carbocyclyl” or “nonaromatic unsaturated carbocyclyl” groups, both of which refer to a nonaromatic carbocycle as defined herein, which contains at least one carbon double bond or one carbon triple bond. 
     As used herein, the term “heterocycloalkyl” or “heterocyclyl” refers to a heteroalicyclic group containing one to four ring heteroatoms each selected from O, S and N. In one embodiment, each heterocycloalkyl group has from 4 to 10 atoms in its ring system, with the proviso that the ring of said group does not contain two adjacent O or S atoms. In another embodiment, the heterocycloalkyl group is fused with an aromatic ring. In one embodiment, the nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen atom may be optionally quaternized. The heterocyclic system may be attached, unless otherwise stated, at any heteroatom or carbon atom that affords a stable structure. A heterocycle may be aromatic or non-aromatic in nature. In one embodiment, the heterocycle is a heteroaryl. 
     An example of a 3-membered heterocycloalkyl group includes, and is not limited to, aziridine. Examples of 4-membered heterocycloalkyl groups include, and are not limited to, azetidine and a beta lactam. Examples of 5-membered heterocycloalkyl groups include, and are not limited to, pyrrolidine, oxazolidine and thiazolidinedione. Examples of 6-membered heterocycloalkyl groups include, and are not limited to, piperidine, morpholine and piperazine. Other non-limiting examples of heterocycloalkyl groups are: 
     
       
         
         
             
             
         
       
     
     Examples of non-aromatic heterocycles include monocyclic groups such as aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, pyrazolidine, imidazoline, dioxolane, sulfolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophane, piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine, morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepin, and hexamethyleneoxide. 
     As used herein, the term “aromatic” refers to a carbocycle or heterocycle with one or more polyunsaturated rings and having aromatic character, i.e. having (4n+2) delocalized i (pi) electrons, where n is an integer. 
     As used herein, the term “aryl,” employed alone or in combination with other terms, means, unless otherwise stated, a carbocyclic aromatic system containing one or more rings (typically one, two or three rings), wherein such rings may be attached together in a pendent manner, such as a biphenyl, or may be fused, such as naphthalene. Examples of aryl groups include phenyl, anthracyl, and naphthyl. 
     As used herein, the term “aryl-(C 1 -C 3 )alkyl” means a functional group wherein a one- to three-carbon alkylene chain is attached to an aryl group, e.g., —CH 2 CH 2 -phenyl. In one embodiment, aryl-(C 1 -C 3 )alkyl is aryl-CH 2 — or aryl-CH(CH 3 )—. The term “substituted aryl-(C 1 -C 3 )alkyl” means an aryl-(C 1 -C 3 )alkyl functional group in which the aryl group is substituted. Similarly, the term “heteroaryl-(C 1 -C 3 )alkyl” means a functional group wherein a one to three carbon alkylene chain is attached to a heteroaryl group, e.g., —CH 2 CH 2 -pyridyl. The term “substituted heteroaryl-(C 1 -C 3 )alkyl” means a heteroaryl-(C 1 -C 3 )alkyl functional group in which the heteroaryl group is substituted. 
     As used herein, the term “heteroaryl” or “heteroaromatic” refers to a heterocycle having aromatic character. A polycyclic heteroaryl may include one or more rings that are partially saturated. Examples include the following moieties: 
     
       
         
         
             
             
         
       
     
     Examples of heteroaryl groups also include pyridyl, pyrazinyl, pyrimidinyl (particularly 2- and 4-pyrimidinyl), pyridazinyl, thienyl, furyl, pyrrolyl (particularly 2-pyrrolyl), imidazolyl, thiazolyl, oxazolyl, pyrazolyl (particularly 3- and 5-pyrazolyl), isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyl and 1,3,4-oxadiazolyl. 
     Examples of polycyclic heterocycles and heteroaryls include indolyl (particularly 3-, 4-, 5-, 6- and 7-indolyl), indolinyl, quinolyl, tetrahydroquinolyl, isoquinolyl (particularly 1- and 5-isoquinolyl), 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl (particularly 2- and 5-quinoxalinyl), quinazolinyl, phthalazinyl, 1,8-naphthyridinyl, 1,4-benzodioxanyl, coumarin, dihydrocoumarin, 1,5-naphthyridinyl, benzofuryl (particularly 3-, 4-, 5-, 6- and 7-benzofuryl), 2,3-dihydrobenzofuryl, 1,2-benzisoxazolyl, benzothienyl (particularly 3-, 4-, 5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (particularly 2-benzothiazolyl and 5-benzothiazolyl), purinyl, benzimidazolyl (particularly 2-benzimidazolyl), benzotriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrrolizidinyl, and quinolizidinyl. 
     As used herein, the term “substituted” means that an atom or group of atoms has replaced hydrogen as the substituent attached to another group. The term “substituted” further refers to any level of substitution, namely mono-, di-, tri-, tetra-, or penta-substitution, where such substitution is permitted. The substituents are independently selected, and substitution may be at any chemically accessible position. In one embodiment, the substituents vary in number between one and four. In another embodiment, the substituents vary in number between one and three. In yet another embodiment, the substituents vary in number between one and two. 
     As used herein, the term “optionally substituted” means that the referenced group may be substituted or unsubstituted. In one embodiment, the referenced group is optionally substituted with zero substituents, i.e., the referenced group is unsubstituted. In another embodiment, the referenced group is optionally substituted with one or more additional group(s) individually and independently selected from groups described herein. 
     In one embodiment, the substituents are independently selected from the group consisting of oxo, halogen, —CN, —NH 2 , —OH, —NH(CH 3 ), —N(CH 3 ) 2 , alkyl (including straight chain, branched and/or unsaturated alkyl), substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, fluoro alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxy, fluoroalkoxy, —S-alkyl, S(═O) 2 alkyl, —C(═O)NH[substituted or unsubstituted alkyl, or substituted or unsubstituted phenyl], —C(═O)N[H or alkyl] 2 , —OC(═O)N[substituted or unsubstituted alkyl] 2 , —NHC(═O)NH[substituted or unsubstituted alkyl, or substituted or unsubstituted phenyl], —NHC(═O)alkyl, —N[substituted or unsubstituted alkyl]C(═O)[substituted or unsubstituted alkyl], —NHC(═O)[substituted or unsubstituted alkyl], —C(OH)[substituted or unsubstituted alkyl] 2 , and —C(NH 2 )[substituted or unsubstituted alkyl] 2 . In another embodiment, by way of example, an optional substituent is selected from oxo, fluorine, chlorine, bromine, iodine, —CN, —NH 2 , —OH, —NH(CH 3 ), —N(CH 3 ) 2 , —CH 3 , —CH 2 CH 3 , —CH(CH 3 ) 2 , —CF 3 , —CH 2 CF 3 , —OCH 3 , —OCH 2 CH 3 , —OCH(CH 3 ) 2 , —OCF 3 , —OCH 2 CF 3 , —S(═O) 2 —CH 3 , —C(═O)NH 2 , —C(═O)—NHCH 3 , —NHC(═O)NHCH 3 , —C(═O)CH 3 , —ON(O) 2 , and —C(═O)OH. In yet one embodiment, the substituents are independently selected from the group consisting of C 1-6  alkyl, —OH, C 1-6  alkoxy, halo, amino, acetamido, oxo and nitro. In yet another embodiment, the substituents are independently selected from the group consisting of C 1-6  alkyl, C 1-6  alkoxy, halo, acetamido, and nitro. As used herein, where a substituent is an alkyl or alkoxy group, the carbon chain may be branched, straight or cyclic. 
     Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range. 
     DESCRIPTION 
     The present invention relates to compositions and methods for treating or preventing a memory-related disease or disorder, such as, but not limited to, PTSD, addiction and addiction-related diseases or disorders. The present invention is based, in part, upon the finding that ACSS2 regulates histone acetylation and neuronal gene transcription. The inhibition of ACSS2 expression (such as by RNA interference) or ACSS2 activity (such as by a small molecule) decreases histone acetylation and impairs long-term spatial memory. Thus, the present invention relates to compositions and method to inhibit ACSS2 in order to inhibit histone acetylation and treat memory-related diseases or disorders. 
     In some embodiments, the composition of the present invention comprises an inhibitor of ACSS2 activity. In some embodiments, the composition comprises an inhibitor of ACSS2 expression. Thus, in various embodiments, the composition comprises an isolated nucleic acid (e.g., siRNA, miRNA, ribozyme, antisense RNA, etc.) that reduces the expression level of ACSS2 in a cell. 
     In some embodiments, the composition comprises an inhibitor of ACSS2 activity. Thus, in various embodiments, the composition comprises a small molecule, nucleic acid, peptide, antibody, antagonist, aptamer, or peptidomimetic that reduces the activity of ACSS2. 
     In some embodiments, the present invention provides a method for treating a neurological or cognitive disease or disorder in a subject. In one embodiment, the neurological or cognitive disease or disorder is a memory-related disease or disorder. In one embodiment, the method comprises administering to a subject an effective amount of a composition comprising an inhibitor of ACSS2. In one embodiment, the method is useful in treating PTSD. 
     In another embodiment, the present invention provides a method for treating addiction or an addiction related disease or disorder in a subject. In some embodiments, the methods of the invention are useful for treating acute alcohol induced memory deficit. In other embodiments, the methods of the invention are useful for treating chronic alcohol induced memory deficit. In some embodiments, the methods comprise administering to a subject an effective amount of a composition comprising an inhibitor of ACSS2. 
     Inhibitors 
     In some embodiments, the present invention provides compositions for treating a neurological or cognitive disease or disorder in a subject. In one embodiment, the neurological or cognitive disease or disorder is a memory-related disease or disorder. In one embodiment, the method comprises administering to a subject an effective amount of a composition comprising an inhibitor of ACSS2. In another embodiment, the present invention provides compositions for treating addiction or an addiction related disease or disorder in a subject. In some embodiments, the methods of the invention are useful for treating acute alcohol induced memory deficit. In other embodiments, the methods of the invention are useful for treating chronic alcohol induced memory deficit. In certain embodiments, the composition inhibits the expression, activity, or both of ACSS2 in the subject. 
     In one embodiment, the composition of the invention comprises an inhibitor of ACSS2. In various embodiments, the inhibitor of ACSS2 is any compound, molecule, or agent that reduces, inhibits, or prevents the expression, activity, or function of ACSS2. Thus, an inhibitor of ACSS2 is any compound, molecule, or agent that reduces ACSS2 expression, activity, or both. In various embodiments, the inhibitor of ACSS2 is a nucleic acid, a peptide, a small molecule, a siRNA, a ribozyme, an antisense nucleic acid, an antagonist, an aptamer, a peptidomimetic, or any combination thereof. 
     Small Molecule Inhibitors 
     In some embodiments, the inhibitor is a small molecule. When the inhibitor is a small molecule, a small molecule may be obtained using standard methods known to the skilled artisan. Such methods include chemical organic synthesis or biological means. Biological means include purification from a biological source, recombinant synthesis and in vitro translation systems, using methods well known in the art. In one embodiment, a small molecule inhibitor of the invention comprises an organic molecule, inorganic molecule, biomolecule, synthetic molecule, and the like. 
     Combinatorial libraries of molecularly diverse chemical compounds potentially useful in treating a variety of diseases and conditions are well known in the art as are method of making the libraries. The method may use a variety of techniques well-known to the skilled artisan including solid phase synthesis, solution methods, parallel synthesis of single compounds, synthesis of chemical mixtures, rigid core structures, flexible linear sequences, deconvolution strategies, tagging techniques, and generating unbiased molecular landscapes for lead discovery vs. biased structures for lead development. 
     In a general method for small library synthesis, an activated core molecule is condensed with a number of building blocks, resulting in a combinatorial library of covalently linked, core-building block ensembles. The shape and rigidity of the core determines the orientation of the building blocks in shape space. The libraries can be biased by changing the core, linkage, or building blocks to target a characterized biological structure (“focused libraries”) or synthesized with less structural bias using flexible cores. 
     The small molecule and small molecule compounds described herein may be present as salts even if salts are not depicted and it is understood that the invention embraces all salts and solvates of the inhibitors depicted here, as well as the non-salt and non-solvate form of the inhibitors, as is well understood by the skilled artisan. In some embodiments, the salts of the inhibitors of the invention are pharmaceutically acceptable salts. 
     Where tautomeric forms may be present for any of the inhibitors described herein, each and every tautomeric form is intended to be included in the present invention, even though only one or some of the tautomeric forms may be explicitly depicted. For example, when a 2-hydroxypyridyl moiety is depicted, the corresponding 2-pyridone tautomer is also intended. 
     The invention also includes any or all of the stereochemical forms, including any enantiomeric or diasteriomeric forms of the inhibitors described. The recitation of the structure or name herein is intended to embrace all possible stereoisomers of inhibitors depicted. All forms of the inhibitors are also embraced by the invention, such as crystalline or non-crystalline forms of the inhibitors. Compositions comprising an inhibitor of the invention are also intended, such as a composition of substantially pure inhibitor, including a specific stereochemical form thereof, or a composition comprising mixtures of inhibitors of the invention in any ratio, including two or more stereochemical forms, such as in a racemic or non-racemic mixture. 
     In one embodiment, the small molecule inhibitor of the invention comprises an analog or derivative of an inhibitor described herein. 
     In one embodiment, the small molecules described herein are candidates for derivatization. As such, in certain instances, the analogs of the small molecules described herein that have modulated potency, selectivity, and solubility are included herein and provide useful leads for drug discovery and drug development. Thus, in certain instances, during optimization new analogs are designed considering issues of drug delivery, metabolism, novelty, and safety. 
     In some instances, small molecule inhibitors described herein are derivatized/analoged as is well known in the art of combinatorial and medicinal chemistry. The analogs or derivatives can be prepared by adding and/or substituting functional groups at various locations. As such, the small molecules described herein can be converted into derivatives/analogs using well known chemical synthesis procedures. For example, all of the hydrogen atoms or substituents can be selectively modified to generate new analogs. Also, the linking atoms or groups can be modified into longer or shorter linkers with carbon backbones or hetero atoms. Also, the ring groups can be changed so as to have a different number of atoms in the ring and/or to include hetero atoms. Moreover, aromatics can be converted to cyclic rings, and vice versa. For example, the rings may be from 5-7 atoms, and may be homocycles or heterocycles. 
     As used herein, the term “analog,” “analogue,” or “derivative” is meant to refer to a chemical compound or molecule made from a parent compound or molecule by one or more chemical reactions. As such, an analog can be a structure having a structure similar to that of the small molecule inhibitors described herein or can be based on a scaffold of a small molecule inhibitor described herein, but differing from it in respect to certain components or structural makeup, which may have a similar or opposite action metabolically. An analog or derivative of any of a small molecule inhibitor in accordance with the present invention can be used to treat an autoimmune disease or disorder. 
     In one embodiment, the small molecule inhibitors described herein can independently be derivatized/analoged by modifying hydrogen groups independently from each other into other substituents. That is, each atom on each molecule can be independently modified with respect to the other atoms on the same molecule. Any traditional modification for producing a derivative/analog can be used. For example, the atoms and substituents can be independently comprised of hydrogen, an alkyl, aliphatic, straight chain aliphatic, aliphatic having a chain hetero atom, branched aliphatic, substituted aliphatic, cyclic aliphatic, heterocyclic aliphatic having one or more hetero atoms, aromatic, heteroaromatic, polyaromatic, polyamino acids, peptides, polypeptides, combinations thereof, halogens, halo-substituted aliphatics, and the like. Additionally, any ring group on a compound can be derivatized to increase and/or decrease ring size as well as change the backbone atoms to carbon atoms or hetero atoms. 
     In one embodiment, the small molecule inhibitor is a compound of Formula (1) 
     
       
         
         
             
             
         
       
     
     wherein, X 11  is selected from the group consisting of C(R 14 )(R 15 ), O, S and NR 15 ; 
     each occurrence of X 12  is selected from the group consisting of C(R 14 )(R 15 ), O, S and NR 15 ; 
     R 11  is selected from the group consisting of hydrogen, —OR 15 , alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein R 11  is optionally substituted; 
     R 12  and R 13  are each independently selected from the group consisting of hydrogen, alkyl, aryl, and heteroaryl, wherein R 12  and R 13  are optionally substituted; 
     each occurrence of R 14  and R 15  are independently selected from the group consisting of hydrogen, halogen, —OH, and alkyl; and 
     n is an integer from 0-8. 
     In one embodiment, n is 0. In one embodiment, n is 1. In one embodiment, n is 2. In one embodiment, n is 3. 
     In one embodiment, R 11  is OR 15 . In one embodiment, R 15  is alkyl. In one embodiment, R 15  is methyl. 
     In one embodiment, R 11  is piperidinyl. 
     In one embodiment, R 11  is morpholinyl. 
     In one embodiment, R 11  is pyrrolidinyl. 
     In one embodiment, R 11  is furanyl. 
     In one embodiment, R 11  is substituted with a hydroxyl group. 
     In one embodiment, R 12  is alkyl. In one embodiment, R 12  is methyl. 
     In one embodiment, R 12  is aryl. In one embodiment, R 12  is phenyl. 
     In one embodiment, R 12  is a C 5 -C 6  heteroaryl. In one embodiment, R 12  is furan. In one embodiment, R 12  is thiophenyl. In one embodiment, R 12  is pyridinyl. 
     In one embodiment, R 13  is alkyl. In one embodiment, R 13  is methyl. 
     In one embodiment, R 13  is aryl. In one embodiment, R 13  is phenyl. 
     In one embodiment, R 13  is a C 5 -C 6  heteroaryl. In one embodiment, R 13  is furan. In one embodiment, R 13  is thiophenyl. In one embodiment, R 13  is pyridinyl. 
     In one embodiment, R 12  and R 13  are the same. 
     In one embodiment, the compound of Formula (1) includes, but is not limited to: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     In one embodiment, the small molecule inhibitor is a compound of Formula (2): 
     
       
         
         
             
             
         
       
     
     wherein, 
     R 21  is selected from the group consisting of —C(R 23 ) m , cycloalkyl, heterocycyl, cycloalkyl-one, and heterocycyl-one; 
     R 22  is selected from the group consisting of alkyl, aryl, heteroaryl, alkyl-aryl, and alkyl-heteroaryl; 
     each occurrence of R 23  is independently selected from the group consisting of hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, heterocycyl, —OH, and —CN; and 
     m is an integer from 1 to 3. 
     In one embodiment, each occurrence of R 23  is independently selected from the group consisting of phenyl, —OH, methyl, ethyl, and —CN. 
     In one embodiment, where m is 3, two occurrences of R 23  are the same and one occurrence of R 23  is different. In one embodiment, where m is 3, each occurrence of R 23  is the same. 
     In one embodiment, R 1  is a group represented by Formula (2a) 
     
       
         
         
             
             
         
       
     
     wherein X 21  is selected from the group consisting of O, N or S; and 
     R 24  is selected from the group consisting of hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, and heterocycyl. 
     In one embodiment, the compound of Formula (2) includes, but is not limited to: 
     
       
         
         
             
             
         
       
     
     In one embodiment, the small molecule inhibitor is a compound of Formula (3) 
     
       
         
         
             
             
         
       
     
     wherein R 31  is selected from the group consisting of —C(R 35 ) p , cycloalkyl, heterocycyl, cycloalkyl-one, heterocycyl-one; 
     R 32  is selected from the group consisting of alkyl, aryl, heteroaryl, —C 1 -C 3  alkyl-(C 3 -C 6  aryl), and —C 1 -C 3  alkyl-(C 3 -C 6  heteroaryl); 
     R 33  and R 34  are each independently selected from the group consisting of hydrogen, halogen, alkyl, aryl, heteroaryl; 
     each occurrence of R 35  is independently selected from the group consisting of hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, heterocycyl, —OH, and —CN; and 
     p is an integer from 1 to 3. 
     In one embodiment, R 32  is ethyl. 
     In one embodiment, R 33  and R 34  are each independently selected from the group consisting of hydrogen and —Cl. 
     In one embodiment R 33  and R 34  are the same. 
     In one embodiment, each occurrence of R 35  is independently selected from the group consisting of phenyl, —OH, methyl, ethyl, and —CN. 
     In one embodiment, where p is 3, two occurrences of R 35  are the same and one occurrence of R 35  is different. In one embodiment, where p is 3, each occurrence of R 35  is the same. 
     In one embodiment, the compound of Formula (3) includes, but is not limited to: 
     
       
         
         
             
             
         
       
     
     In one embodiment, the small molecule inhibitor is a compound of Formula (4): 
     
       
         
         
             
             
         
       
     
     wherein, 
     X 41  is selected from the group consisting of O and S; 
     R 41  is selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and combinations thereof, wherein R 41  may be optionally substituted; and 
     R 42  and R 43  are each independently selected from the group consisting of phenyl, thiophenyl and furanyl. 
     In one embodiment, R 42  and R 43  are the same. 
     In one embodiment, R 41  is adamantyl. 
     In one embodiment, R 41  is piperidinyl. 
     In one embodiment, R 41  is morpholinyl. 
     In one embodiment, R 41  is pyrrolidinyl. 
     In one embodiment, R 41  is furanyl. 
     In one embodiment, R 41  is alkyl. In one embodiment, R 41  is C 1 -C 25  alkyl. In one embodiment, the alkyl is a branched chain alkyl. In one embodiment, the alkyl is a straight chain alkyl. 
     In one embodiment, R 21  is —C 3 -C 10  cycloalkyl, which may be optionally substituted. In one embodiment, the cycloalkyl group is substituted. In one embodiment, the cycloalkyl group is unsubstituted. In one embodiment, the cycloalkyl group is monocyclic. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, and the like. In another embodiment, the cycloalkyl group is polycyclic. For example, a polycyclic cycloalkyl group may be formed by joining two or more —C 3 -C 10  cycloalkyl groups. Non-limiting examples of polycyclic cycloalkyl groups include adamantane and norbornane. In one embodiment, the cycloalkyl group is adamantane, which may be optionally substituted. Cycloalkyl groups may also be dicyclic including, but not limited to, tetrahydronaphthyl, indanyl, and tetrahydropentalene. In one embodiment, the cycloalkyl group is saturated or partially unsaturated. Non-limiting examples of saturated or partially unsaturated cycloalkyl groups include cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptatrienyl, cyclooctenyl, cycloocta-dienyl, cyclooctatrienyl, cyclooctatetraenyl, cyclononenyl, cyclononadienyl, cyclodecenyl, cyclodekadienyl, cyclooctynyl, cyclononynyl, cyclodecynyl, and the like. In one embodiment, the cycloalkyl group is fused with an aromatic ring 
     In one embodiment, the compound of formula (4) is selected from the group consisting of 
     
       
         
         
             
             
         
       
     
     Preparation of the Small Molecule Inhibitors of the Invention 
     Compounds of Formulae (1)-(4) may be prepared by the general schemes described herein, using the synthetic method known by those skilled in the art. The following examples illustrate non-limiting embodiments of the invention. 
     In a non-limiting embodiment, the synthesis of compounds of Formulae (1) and (4) is accomplished by treating 4-nitro-o-phenylenediamine (a) with a diketone (b) to form a 6-nitroquinoxaline (c), which is subsequently reduced via Pd/C-catalyzed hydrogenation to produce a 6-aminoquinoxaline (d). A diketone (a) can be produced using a method known in the art (Tet. Lett., 1995, 36:7305-7308, which is incorporated herein by reference in its entirety. 
     
       
         
         
             
             
         
       
     
     Quinoxaline d is then treated with an isocyanate to form a compound of Formulae (1) or (4). 
     
       
         
         
             
             
         
       
     
     In another non-limiting embodiment, quinoxaline d is first treated with triphosgene, followed by the addition of an amine, to form a compound of Formulae (1) or (4). 
     
       
         
         
             
             
         
       
     
     The compounds of the invention may possess one or more stereocenters, and each stereocenter may exist independently in either the R or S configuration. In one embodiment, compounds described herein are present in optically active or racemic forms. It is to be understood that the compounds described herein encompass racemic, optically-active, regioisomeric and stereoisomeric forms, or combinations thereof that possess the therapeutically useful properties described herein. Preparation of optically active forms is achieved in any suitable manner, including by way of non-limiting example, by resolution of the racemic form with recrystallization techniques, synthesis from optically-active starting materials, chiral synthesis, or chromatographic separation using a chiral stationary phase. In one embodiment, a mixture of one or more isomers is utilized as the therapeutic compound described herein. In another embodiment, compounds described herein contain one or more chiral centers. These compounds are prepared by any means, including stereoselective synthesis, enantioselective synthesis and/or separation of a mixture of enantiomers and/or diastereomers. Resolution of compounds and isomers thereof is achieved by any means including, by way of non-limiting example, chemical processes, enzymatic processes, fractional crystallization, distillation, and chromatography. 
     The methods and formulations described herein include the use of N-oxides (if appropriate), crystalline forms (also known as polymorphs), solvates, amorphous phases, and/or pharmaceutically acceptable salts of compounds having the structure of any compound of the invention, as well as metabolites and active metabolites of these compounds having the same type of activity. Solvates include water, ether (e.g., tetrahydrofuran, methyl tert-butyl ether) or alcohol (e.g., ethanol) solvates, acetates and the like. In one embodiment, the compounds described herein exist in solvated forms with pharmaceutically acceptable solvents such as water and ethanol. In another embodiment, the compounds described herein exist in unsolvated form. 
     In one embodiment, the compounds of the invention may exist as tautomers. All tautomers are included within the scope of the compounds presented herein. 
     Compounds described herein also include isotopically-labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds described herein include and are not limited to  2 H,  3 H,  11 C,  13 C,  14 C,  36 Cl,  18 F,  123 I,  125 I,  13 N,  15 N,  15 O,  17 O,  18 O,  32 P, and  35 S. Isotopically-labeled compounds are prepared by any suitable method or by processes using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed. 
     In one embodiment, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels. 
     The compounds described herein, and other related compounds having different substituents are synthesized using techniques and materials described herein and as described, for example, in Fieser &amp; Fieser&#39;s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd&#39;s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), Larock&#39;s Comprehensive Organic Transformations (VCH Publishers Inc., 1989), March, Advanced Organic Chemistry 4 th  Ed., (Wiley 1992); Carey &amp; Sundberg, Advanced Organic Chemistry 4th Ed., Vols. A and B (Plenum 2000,2001), and Green &amp; Wuts, Protective Groups in Organic Synthesis 3rd Ed., (Wiley 1999) (all of which are incorporated by reference for such disclosure). General methods for the preparation of compound as described herein are modified by the use of appropriate reagents and conditions, for the introduction of the various moieties found in the formula as provided herein. 
     Compounds described herein are synthesized using any suitable procedures starting from compounds that are available from commercial sources, or are prepared using procedures described herein. 
     In one embodiment, reactive functional groups, such as hydroxyl, amino, imino, thio or carboxy groups, are protected in order to avoid their unwanted participation in reactions. Protecting groups are used to block some or all of the reactive moieties and prevent such groups from participating in chemical reactions until the protective group is removed. In another embodiment, each protective group is removable by a different means. Protective groups that are cleaved under totally disparate reaction conditions fulfill the requirement of differential removal. 
     In one embodiment, protective groups are removed by acid, base, reducing conditions (such as, for example, hydrogenolysis), and/or oxidative conditions. Groups such as trityl, dimethoxytrityl, acetal and t-butyldimethylsilyl are acid labile and are used to protect carboxy and hydroxy reactive moieties in the presence of amino groups protected with Cbz groups, which are removable by hydrogenolysis, and Fmoc groups, which are base labile. Carboxylic acid and hydroxy reactive moieties are blocked with base labile groups such as, but not limited to, methyl, ethyl, and acetyl, in the presence of amines that are blocked with acid labile groups, such as t-butyl carbamate, or with carbamates that are both acid and base stable but hydrolytically removable. 
     In one embodiment, carboxylic acid and hydroxy reactive moieties are blocked with hydrolytically removable protective groups such as the benzyl group, while amine groups capable of hydrogen bonding with acids are blocked with base labile groups such as Fmoc. Carboxylic acid reactive moieties are protected by conversion to simple ester compounds as exemplified herein, which include conversion to alkyl esters, or are blocked with oxidatively-removable protective groups such as 2,4-dimethoxybenzyl, while co-existing amino groups are blocked with fluoride labile silyl carbamates. 
     Allyl blocking groups are useful in the presence of acid- and base-protecting groups since the former are stable and are subsequently removed by metal or pi-acid catalysts. For example, an allyl-blocked carboxylic acid is deprotected with a palladium-catalyzed reaction in the presence of acid labile t-butyl carbamate or base-labile acetate amine protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate is attached. As long as the residue is attached to the resin, that functional group is blocked and does not react. Once released from the resin, the functional group is available to react. 
     Typically blocking/protecting groups may be selected from: 
     
       
         
         
             
             
         
       
     
     Other protecting groups, plus a detailed description of techniques applicable to the creation of protecting groups and their removal are described in Greene &amp; Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley &amp; Sons, New York, N.Y., 1999, and Kocienski, Protective Groups, Thieme Verlag, New York, N.Y., 1994, which are incorporated herein by reference for such disclosure. 
     Nucleic Acid Inhibitors 
     In some embodiments, the inhibitor is nucleic acid. In various embodiments, the inhibitor is an siRNA, miRNA, shRNA, or an antisense molecule, which inhibits ACSS2. In one embodiment, the nucleic acid comprises a promoter/regulatory sequence such that the nucleic acid is capable of directing expression of the inhibitor nucleic acid. Thus, the invention encompasses expression vectors and methods for the introduction of exogenous DNA into cells with concomitant expression of the exogenous DNA in the cells such as those described, for example, in Sambrook et al. (2012, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in Ausubel et al. (1997, Current Protocols in Molecular Biology, John Wiley &amp; Sons, New York) and as described elsewhere herein. 
     In another aspect of the invention, ACSS2, can be inhibited by way of inactivating and/or sequestering ACSS2. As such, inhibiting the activity of ACSS2 can be accomplished by using a transdominant negative mutant. 
     In one embodiment, siRNA is used to decrease the level of ACSS2 protein. RNA interference (RNAi) is a phenomenon in which the introduction of double-stranded RNA (dsRNA) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. In the cell, long dsRNAs are cleaved into short 21-25 nucleotide small interfering RNAs, or siRNAs, by a ribonuclease known as Dicer. The siRNAs subsequently assemble with protein components into an RNA-induced silencing complex (RISC), unwinding in the process. Activated RISC then binds to complementary transcript by base pairing interactions between the siRNA antisense strand and the mRNA. The bound mRNA is cleaved and sequence specific degradation of mRNA results in gene silencing. See, for example, U.S. Pat. No. 6,506,559; Fire et al., 1998, Nature 391(19):306-311; Timmons et al., 1998, Nature 395:854; Montgomery et al., 1998, TIG 14 (7):255-258; David R. Engelke, Ed., RNA Interference (RNAi) Nuts &amp; Bolts of RNAi Technology, DNA Press, Eagleville, P A (2003); and Gregory J. Hannon, Ed., RNAi A Guide to Gene Silencing, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2003). Soutschek et al. (2004, Nature 432:173-178) describe a chemical modification to siRNAs that aids in intravenous systemic delivery. Optimizing siRNAs involves consideration of overall G/C content, C/T content at the termini, Tm and the nucleotide content of the 3′ overhang. See, for instance, Schwartz et al., 2003, Cell, 115:199-208 and Khvorova et al., 2003, Cell 115:209-216. Therefore, the present invention also includes methods of decreasing levels of ACSS2 using RNAi technology. 
     In another aspect, the invention includes a vector comprising an siRNA or antisense nucleic acid. In one embodiment, the siRNA or antisense polynucleotide is capable of inhibiting the expression of a target polypeptide, wherein the target polypeptide is ACSS2. The incorporation of a desired polynucleotide into a vector and the choice of vectors is well-known in the art as described in, for example, Sambrook et al. (2012), and in Ausubel et al. (1997), and elsewhere herein. 
     In certain embodiments, the expression vectors described herein encode a short hairpin RNA (shRNA) inhibitor. shRNA inhibitors are well known in the art and are directed against the mRNA of a target, thereby decreasing the expression of the target. In certain embodiments, the encoded shRNA is expressed by a cell, and is then processed into siRNA. For example, in certain instances, the cell possesses native enzymes (e.g., dicer) that cleaves the shRNA to form siRNA. 
     The siRNA, shRNA, or antisense nucleic acid can be cloned into a number of types of vectors as described elsewhere herein. For expression of the siRNA or antisense polynucleotide, at least one module in each promoter functions to position the start site for RNA synthesis. 
     In order to assess the expression of the siRNA, shRNA, or antisense nucleic, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from the population of cells sought to be transfected or infected using a viral vector. In other embodiments, the selectable marker may be carried on a separate piece of DNA and used in a co-transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers are known in the art and include, for example, antibiotic-resistance genes, such as neomycin resistance and the like. 
     Therefore, in another aspect, the invention relates to a vector, comprising the nucleotide sequence of the invention or the construct of the invention. The choice of the vector will depend on the host cell in which it is to be subsequently introduced. In a particular embodiment, the vector of the invention is an expression vector. Suitable host cells include a wide variety of prokaryotic and eukaryotic host cells. In specific embodiments, the expression vector is selected from the group consisting of a viral vector, a bacterial vector and a mammalian cell vector. Prokaryote- and/or eukaryote-vector based systems can be employed for use with the present invention to produce polynucleotides, or their cognate polypeptides. Many such systems are commercially and widely available. 
     Further, the expression vector may be provided to a cell in the form of a viral vector. Viral vector technology is well known in the art and is described, for example, in Sambrook et al. (2012), and in Ausubel et al. (1997), and in other virology and molecular biology manuals. Viruses, which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers. (See, e.g., WO 01/96584; WO 01/29058; and U.S. Pat. No. 6,326,193. 
     By way of illustration, the vector in which the nucleic acid sequence is introduced can be a plasmid, which is or is not integrated in the genome of a host cell when it is introduced in the cell. Illustrative, non-limiting examples of vectors in which the nucleotide sequence of the invention or the gene construct of the invention can be inserted include a tet-on inducible vector for expression in eukaryote cells. 
     The vector may be obtained by conventional methods known by persons skilled in the art (Sambrook et al., 2012). In a particular embodiment, the vector is a vector useful for transforming animal cells. 
     In one embodiment, the recombinant expression vectors may also contain nucleic acid molecules, which encode a peptide or peptidomimetic inhibitor of invention, described elsewhere herein. 
     A promoter may be one naturally associated with a gene or polynucleotide sequence, as may be obtained by isolating the 5′ non-coding sequences located upstream of the coding segment and/or exon. Such a promoter can be referred to as “endogenous.” Similarly, an enhancer may be one naturally associated with a polynucleotide sequence, located either downstream or upstream of that sequence. Alternatively, certain advantages will be gained by positioning the coding polynucleotide segment under the control of a recombinant or heterologous promoter, which refers to a promoter that is not normally associated with a polynucleotide sequence in its natural environment. A recombinant or heterologous enhancer refers also to an enhancer not normally associated with a polynucleotide sequence in its natural environment. Such promoters or enhancers may include promoters or enhancers of other genes, and promoters or enhancers isolated from any other prokaryotic, viral, or eukaryotic cell, and promoters or enhancers not “naturally occurring,” i.e., containing different elements of different transcriptional regulatory regions, and/or mutations that alter expression. In addition to producing nucleic acid sequences of promoters and enhancers synthetically, sequences may be produced using recombinant cloning and/or nucleic acid amplification technology, including PCR, in connection with the compositions disclosed herein (U.S. Pat. Nos. 4,683,202, 5,928,906). Furthermore, it is contemplated the control sequences that direct transcription and/or expression of sequences within non-nuclear organelles such as mitochondria, chloroplasts, and the like, can be employed as well. 
     Naturally, it will be important to employ a promoter and/or enhancer that effectively directs the expression of the DNA segment in the cell type, organelle, and organism chosen for expression. Those of skill in the art of molecular biology generally know how to use promoters, enhancers, and cell type combinations for protein expression, for example, see Sambrook et al. (2012). The promoters employed may be constitutive, tissue-specific, inducible, and/or useful under the appropriate conditions to direct high level expression of the introduced DNA segment, such as is advantageous in the large-scale production of recombinant proteins and/or peptides. The promoter may be heterologous or endogenous. 
     The recombinant expression vectors may also contain a selectable marker gene, which facilitates the selection of transformed or transfected host cells. Suitable selectable marker genes are genes encoding proteins such as G418 and hygromycin, which confer resistance to certain drugs, 0-galactosidase, chloramphenicol acetyltransferase, firefly luciferase, or an immunoglobulin or portion thereof such as the Fc portion of an immunoglobulin, for example, IgG. The selectable markers may be introduced on a separate vector from the nucleic acid of interest. 
     Following the generation of the siRNA polynucleotide, a skilled artisan will understand that the siRNA polynucleotide will have certain characteristics that can be modified to improve the siRNA as a therapeutic compound. Therefore, the siRNA polynucleotide may be further designed to resist degradation by modifying it to include phosphorothioate, or other linkages, methylphosphonate, sulfone, sulfate, ketyl, phosphorodithioate, phosphoramidate, phosphate esters, and the like (see, e.g., Agrwal et al., 1987, Tetrahedron Lett. 28:3539-3542; Stec et al., 1985 Tetrahedron Lett. 26:2191-2194; Moody et al., 1989 Nucleic Acids Res. 12:4769-4782; Eckstein, 1989 Trends Biol. Sci. 14:97-100; Stein, In: Oligodeoxynucleotides. Antisense Inhibitors of Gene Expression, Cohen, ed., Macmillan Press, London, pp. 97-117 (1989)). 
     Any polynucleotide may be further modified to increase its stability in vivo. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5′ and/or 3′ ends; the use of phosphorothioate or 2′ O-methyl rather than phosphodiester linkages in the backbone; and/or the inclusion of nontraditional bases such as inosine, queosine, and wybutosine and the like, as well as acetyl-methyl-, thio- and other modified forms of adenine, cytidine, guanine, thymine, and uridine. 
     In one embodiment of the invention, an antisense nucleic acid sequence, which is expressed by a plasmid vector is used to inhibit ACSS2 protein expression. The antisense expressing vector is used to transfect a mammalian cell or the mammal itself, thereby causing reduced endogenous expression of ACSS2. 
     Antisense molecules and their use for inhibiting gene expression are well known in the art (see, e.g., Cohen, 1989, In: Oligodeoxyribonucleotides, Antisense Inhibitors of Gene Expression, CRC Press). Antisense nucleic acids are DNA or RNA molecules that are complementary, as that term is defined elsewhere herein, to at least a portion of a specific mRNA molecule (Weintraub, 1990, Scientific American 262:40). In the cell, antisense nucleic acids hybridize to the corresponding mRNA, forming a double-stranded molecule thereby inhibiting the translation of genes. 
     The use of antisense methods to inhibit the translation of genes is known in the art, and is described, for example, in Marcus-Sakura (1988, Anal. Biochem. 172:289). Such antisense molecules may be provided to the cell via genetic expression using DNA encoding the antisense molecule as taught by Inoue, 1993, U.S. Pat. No. 5,190,931. 
     Alternatively, antisense molecules of the invention may be made synthetically and then provided to the cell. In one embodiment, the antisense oligomers are between about 10 to about 30 nucleotides. In one embodiment, the antisense oligomers are about 15 nucleotides. In one embodiment, antisense oligomers of about 10 to about 30 nucleotides are easily synthesized and introduced into a target cell. Synthetic antisense molecules contemplated by the invention include oligonucleotide derivatives known in the art which have improved biological activity compared to unmodified oligonucleotides (see U.S. Pat. No. 5,023,243). 
     In one embodiment of the invention, a ribozyme is used to inhibit ACSS2 protein expression. Ribozymes useful for inhibiting the expression of a target molecule may be designed by incorporating target sequences into the basic ribozyme structure, which are complementary, for example, to the mRNA sequence encoding ACSS2. Ribozymes targeting ACSS2, may be synthesized using commercially available reagents (Applied Biosystems, Inc., Foster City, Calif.) or they may be genetically expressed from DNA encoding them. 
     In one embodiment, the inhibitor of ACSS2 may comprise one or more components of a CRISPR-Cas system. CRISPR methodologies employ a nuclease, CRISPR-associated (Cas), that complexes with small RNAs as guides (gRNAs) to cleave DNA in a sequence-specific manner upstream of the protospacer adjacent motif (PAM) in any genomic location. CRISPR may use separate guide RNAs known as the crRNA and tracrRNA. These two separate RNAs have been combined into a single RNA to enable site-specific mammalian genome cutting through the design of a short guide RNA. Cas and guide RNA (gRNA) may be synthesized by known methods. Cas/guide-RNA (gRNA) uses a non-specific DNA cleavage protein Cas, and an RNA oligo to hybridize to target and recruit the Cas/gRNA complex. In one embodiment, a guide RNA (gRNA) targeted to a gene encoding ACSS2, and a CRISPR-associated (Cas) peptide form a complex to induce mutations within the targeted gene. In one embodiment, the inhibitor comprises a gRNA or a nucleic acid molecule encoding a gRNA. In one embodiment, the inhibitor comprises a Cas peptide or a nucleic acid molecule encoding a Cas peptide. 
     Polypeptide Inhibitors 
     In some embodiments, the inhibitor is a peptide or polypeptide inhibitor that inhibits ACSS2. For example, in one embodiment, the peptide inhibitor of the invention inhibits ACSS2 directly by binding to ACSS2 thereby preventing the normal functional activity of ACSS2. In another embodiment, the peptide inhibitor of the invention inhibits ACSS2 by competing with endogenous ACSS2. In yet another embodiment, the peptide inhibitor of the invention inhibits the activity of ACSS2 by acting as a transdominant negative mutant. 
     Variants of the peptides and polypeptides according to the present invention may be (i) one in which one or more of the amino acid residues are substituted with a conserved or non-conserved amino acid residue and such substituted amino acid residue may or may not be one encoded by the genetic code, (ii) one in which there are one or more modified amino acid residues, e.g., residues that are modified by the attachment of substituent groups, (iii) one in which the polypeptide is an alternative splice variant of the polypeptide of the present invention, (iv) fragments of the polypeptides and/or (v) one in which the polypeptide is fused with another polypeptide, such as a leader or secretory sequence or a sequence which is employed for purification (for example, His-tag) or for detection (for example, Sv5 epitope tag). The fragments include polypeptides generated via proteolytic cleavage (including multi-site proteolysis) of an original sequence. Variants may be post-translationally, or chemically modified. Such variants are deemed to be within the scope of those skilled in the art from the teaching herein. 
     Antibody Inhibitors 
     In some embodiments, the inhibitor is an antibody, or antibody fragment. In some embodiments, the inhibitor is an antibody, or antibody fragment, that specifically binds to ACSS2. That is, the antibody can inhibit ACSS2 to provide a beneficial effect. 
     The antibodies may be intact monoclonal or polyclonal antibodies, and immunologically active fragments (e.g., a Fab or (Fab) 2  fragment), an antibody heavy chain, an antibody light chain, humanized antibodies, a genetically engineered single chain Fv molecule (Ladner et al, U.S. Pat. No. 4,946,778), or a chimeric antibody, for example, an antibody which contains the binding specificity of a murine antibody, but in which the remaining portions are of human origin. Antibodies including monoclonal and polyclonal antibodies, humanized antibodies, fragments and chimeras, may be prepared using methods known to those skilled in the art. 
     The antibody may comprise a heavy chain and a light chain complementarity determining region (“CDR”) set, respectively interposed between a heavy chain and a light chain framework (“FR”) set which provide support to the CDRs and define the spatial relationship of the CDRs relative to each other. The CDR set may contain three hypervariable regions of a heavy or light chain V region. Proceeding from the N-terminus of a heavy or light chain, these regions are denoted as “CDR1,” “CDR2,” and “CDR3,” respectively. An antigen-binding site, therefore, may include six CDRs, comprising the CDR set from each of a heavy and a light chain V region. 
     The antibody can be an immunoglobulin (Ig). The Ig can be, for example, IgA, IgM, IgD, IgE, and IgG. The immunoglobulin can include the heavy chain polypeptide and the light chain polypeptide. The heavy chain polypeptide of the immunoglobulin can include a VH region, a CH1 region, a hinge region, a CH2 region, and a CH3 region. The light chain polypeptide of the immunoglobulin can include a VL region and CL region. 
     The antibody can be a polyclonal or monoclonal antibody. The antibody can be a chimeric antibody, a single chain antibody, an affinity matured antibody, a human antibody, a humanized antibody, or a fully human antibody. The humanized antibody can be an antibody from a non-human species that binds the desired antigen having one or more complementarity determining regions (CDRs) from the non-human species and framework regions from a human immunoglobulin molecule. 
     The antibody can be a bispecific antibody. The bispecific antibody can bind or react with two antigens, for example, two of the antigens described below in more detail. The bispecific antibody can be comprised of fragments of two of the antibodies described herein, thereby allowing the bispecific antibody to bind or react with two desired target molecules, which may include the antigen, which is described below in more detail, a ligand, including a ligand for a receptor, a receptor, including a ligand-binding site on the receptor, a ligand-receptor complex, and a marker. Bispecific antibodies can comprise a first antigen-binding site that specifically binds to a first target and a second antigen-binding site that specifically binds to a second target, with particularly advantageous properties such as producibility, stability, binding affinity, biological activity, specific targeting of certain T cells, targeting efficiency and reduced toxicity. In some instances, there are bispecific antibodies, wherein the bispecific antibody binds to the first target with high affinity and to the second target with low affinity. In other instances, there are bispecific antibodies, wherein the bispecific antibody binds to the first target with low affinity and to the second target with high affinity. In other instances, there are bispecific antibodies, wherein the bispecific antibody binds to the first target with a desired affinity and to the second target with a desired affinity. 
     Antibodies can be prepared using intact polypeptides or fragments containing an immunizing antigen of interest. The polypeptide or oligopeptide used to immunize an animal may be obtained from the translation of RNA or synthesized chemically and can be conjugated to a carrier protein, if desired. Suitable carriers that may be chemically coupled to peptides include bovine serum albumin and thyroglobulin, keyhole limpet hemocyanin. The coupled polypeptide may then be used to immunize the animal (e.g., a mouse, a rat, or a rabbit). 
     Combinations 
     In some embodiments, the compositions of the present invention comprise a combination of ACSS2 inhibitors described herein. In certain embodiments, a composition comprising a combination of inhibitors described herein has an additive effect, wherein the overall effect of the combination is approximately equal to the sum of the effects of each individual inhibitor. In other embodiments, a composition comprising a combination of inhibitors described herein has a synergistic effect, wherein the overall effect of the combination is greater than the sum of the effects of each individual inhibitor. 
     In some embodiments, the composition of the present invention comprises a combination of an ACSS2 inhibitor and second therapeutic agent. For example, in one embodiment the second therapeutic agents include, but are not limited to, a PTSD treatment, an anxiety treatment, and a substance abuse treatment. 
     In some embodiments, the second therapeutic is a PTSD treatment. Exemplary therapeutics include, but are not limited to, anti-anxiety treatments, antidepressants, and adrenergic agents. In one embodiment, the PTSD treatment is a therapy treatment. For example, in one embodiment the PTSD treatment includes, psychotherapy, behavioral or cognitive behavioral therapy, eye movement desensitization and reprocessing (EMDR) group therapy, transcranial magnetic stimulation, deep brain stimulation and neurofeedback techniques, and medications including ketamine and d-cycloserine. 
     In one embodiment, administration of the ACSS2 inhibitor in the emergency room or in intensive care units can be used for PTSD prophylaxis. In the peritraumatic phase, reactivated memory traces are vulnerable to disruption, thus ACSS2 inhibition offers the potential to affect reconsolidation of trauma memories. 
     In some embodiments, the second therapeutic is a substance abuse treatment. For example, in one embodiment the substance abuse treatment includes, but is not limited to, naltrexone, disulfiram, acamprosate, topiramate, nicotine replacement therapy, nicotinic receptor antagonists, nicotinic receptor partial agonists, suboxone, levomethadyl acetate, dihydrocodeine, buprenorphine, ketamine, methadone, and dihydroetorphine. 
     A composition comprising a combination of inhibitors comprises individual inhibitors in any suitable ratio. For example, in one embodiment, the composition comprises a 1:1 ratio of two individual inhibitors. However, the combination is not limited to any particular ratio. Rather any ratio that is shown to be effective is encompassed. 
     Methods 
     In some embodiments, the invention provides methods of inhibiting the ACSS2 in a subject in need thereof. In one embodiment, the method comprises administering to the subject an effective amount of a composition comprising an ACSS2 inhibitor. 
     In one embodiment, the invention provides a method for modulating chromatin acetylation in a subject. In one embodiment, the chromatin acetylation is histone acetylation. In one embodiment, the chromatin is neural chromatin. In one embodiment, methods of the invention modulate neuronal plasticity in a subject. In one embodiment, the method comprises administering to a subject an effective amount of a composition comprising an inhibitor of ACSS2. In one embodiment, the inhibitor of ACSS2 decreases histone acetylation. 
     In one aspect, the present invention provides a method for treating neurological or cognitive disease or disorder in a subject. In one embodiment, the neurological or cognitive disease or disorder is a memory-related disease or disorder. In one embodiment, the neurological or cognitive disease or disorder is a neuropsychiatric disorder. For example, in one embodiment the neuropsychiatric disorder includes, but is not limited to, anxiety disorders, psychotic disorders, mood disorders and somatoform disorders. 
     Exemplary neurological or cognitive diseases or disorders include, but are not limited to, post-traumatic stress disorder (PTSD), bipolar disorder, depression, Tourette&#39;s Syndrome, schizophrenia, obsessive-compulsive disorder, generalized anxiety disorder, panic disorders, phobias, personality disorders, including antisocial personality disorder, and other disorders involving troubling memories. In one embodiment, the neurological or cognitive diseases or disorders is PTSD. 
     In another embodiment, the present invention provides a method for treating addiction or an addiction related disease or disorder in a subject. In one embodiment, the addiction includes, but is not limited to, addiction to: alcohol, tobacco, opioids, sedatives, hypnotics, anxiolytics, cocaine, cannabis, amphetamines, hallucinogens, inhalants, phencyclidine, impulse control disorders and behavioral addictions. 
     In one embodiment, the addiction is an alcohol addiction. In one embodiment, the method of the invention treats acute and/or chronic alcohol induced memory deficit. 
     In one embodiment, the invention provides a method for treating alcohol-related memory and cue-induced craving in augmented psychotherapy. In one embodiment, the method comprises administering to a subject an effective amount of a composition comprising an inhibitor of ACSS2. In one embodiment, the inhibitor of ACSS2 decreases histone acetylation. 
     In one embodiment, the method comprises administering to the subject an effective amount of a composition that reduces or inhibits the expression or activity of ACSS2. 
     One of skill in the art will appreciate that the inhibitors of the invention can be administered singly or in any combination. Further, the inhibitors of the invention can be administered singly or in any combination in a temporal sense, in that they may be administered concurrently, or before, and/or after each other. One of ordinary skill in the art will appreciate, based on the disclosure provided herein, that the inhibitor compositions of the invention can be used to prevent or to treat an autoimmune disease or disorder, and that an inhibitor composition can be used alone or in any combination with another modulator to affect a therapeutic result. In various embodiments, any of the inhibitor compositions of the invention described herein can be administered alone or in combination with other modulators of other molecules associated with autoimmune diseases. 
     In one embodiment, the invention includes a method comprising administering a combination of inhibitors described herein. In certain embodiments, the method has an additive effect, wherein the overall effect of the administering a combination of inhibitors is approximately equal to the sum of the effects of administering each individual inhibitor. In other embodiments, the method has a synergistic effect, wherein the overall effect of administering a combination of inhibitors is greater than the sum of the effects of administering each individual inhibitor. 
     The method comprises administering a combination of inhibitors in any suitable ratio. For example, in one embodiment, the method comprises administering two individual inhibitors at a 1:1 ratio. However, the method is not limited to any particular ratio. Rather any ratio that is shown to be effective is encompassed. 
     Pharmaceutical Compositions and Formulations 
     The invention also encompasses the use of pharmaceutical compositions of the invention or salts thereof to practice the methods of the invention. Such a pharmaceutical composition may consist of at least one modulator (e.g., inhibitor) composition of the invention or a salt thereof in a form suitable for administration to a subject, or the pharmaceutical composition may comprise at least one modulator (e.g., inhibitor) composition of the invention or a salt thereof, and one or more pharmaceutically acceptable carriers, one or more additional ingredients, or some combination of these. The compound of the invention may be present in the pharmaceutical composition in the form of a physiologically acceptable salt, such as in combination with a physiologically acceptable cation or anion, as is well known in the art. 
     In an embodiment, the pharmaceutical compositions useful for practicing the methods of the invention may be administered to deliver a dose of between 1 ng/kg/day and 100 mg/kg/day. In another embodiment, the pharmaceutical compositions useful for practicing the invention may be administered to deliver a dose of between 1 ng/kg/day and 500 mg/kg/day. 
     The relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w/w) active ingredient. 
     Pharmaceutical compositions that are useful in the methods of the invention may be suitably developed for oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, ophthalmic, or another route of administration. A composition useful within the methods of the invention may be directly administered to the skin, or any other tissue of a mammal. Other contemplated formulations include liposomal preparations, resealed erythrocytes containing the active ingredient, and immunologically-based formulations. The route(s) of administration will be readily apparent to the skilled artisan and will depend upon any number of factors including the type and severity of the disease being treated, the type and age of the veterinary or human subject being treated, and the like. 
     The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit. 
     As used herein, a “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient that would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage. The unit dosage form may be for a single daily dose or one of multiple daily doses (e.g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose. 
     In one embodiment, the compositions of the invention are formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of a compound or conjugate of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers that are useful, include, but are not limited to, glycerol, water, saline, ethanol and other pharmaceutically acceptable salt solutions such as phosphates and salts of organic acids. Examples of these and other pharmaceutically acceptable carriers are described in Remington&#39;s Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey). 
     The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, are included in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate or gelatin. In one embodiment, the pharmaceutically acceptable carrier is not DMSO alone. 
     Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, vaginal, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration, known to the art. The pharmaceutical preparations may be sterilized and if desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and/or aromatic substances and the like. They may also be combined where desired with other active agents, e.g., other analgesic agents. 
     As used herein, “additional ingredients” include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials. Other “additional ingredients” that may be included in the pharmaceutical compositions of the invention are known in the art and described, for example in Genaro, ed. (1985, Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.), which is incorporated herein by reference. 
     The composition of the invention may comprise a preservative from about 0.005% to 2.0% by total weight of the composition. The preservative is used to prevent spoilage in the case of exposure to contaminants in the environment. Examples of preservatives useful in accordance with the invention included but are not limited to those selected from the group consisting of benzyl alcohol, sorbic acid, parabens, imidurea and combinations thereof. An exemplary preservative is a combination of about 0.5% to 2.0% benzyl alcohol and 0.05% to 0.5% sorbic acid. 
     In one embodiment, the composition includes an anti-oxidant and a chelating agent that inhibits the degradation of the compound. Exemplary antioxidants for some compounds include BHT, BHA, alpha-tocopherol and ascorbic acid in the preferred range of about 0.01% to 0.3%. In one embodiment, the antioxidant is BHT in the range of 0.03% to 0.1% by weight by total weight of the composition. In one embodiment, the chelating agent is present in an amount of from 0.01% to 0.5% by weight by total weight of the composition. Exemplary chelating agents include edetate salts (e.g. disodium edetate) and citric acid in the weight range of about 0.01% to 0.20%. In one embodiment, the chelating agent is in the range of 0.02% to 0.10% by weight by total weight of the composition. The chelating agent is useful for chelating metal ions in the composition that may be detrimental to the shelf life of the formulation. While BHT and disodium edetate are exemplary antioxidant and chelating agents, respectively, for some compounds, other suitable and equivalent antioxidants and chelating agents may be substituted therefore as would be known to those skilled in the art. 
     Liquid suspensions may be prepared using conventional methods to achieve suspension of the active ingredient in an aqueous or oily vehicle. Aqueous vehicles include, for example, water, and isotonic saline. Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as  arachis , olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Liquid suspensions may further comprise one or more additional ingredients including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavorings, coloring agents, and sweetening agents. Oily suspensions may further comprise a thickening agent. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum acacia, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose. Known dispersing or wetting agents include, but are not limited to, naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with a fatty acid, with a long chain aliphatic alcohol, with a partial ester derived from a fatty acid and a hexitol, or with a partial ester derived from a fatty acid and a hexitol anhydride (e.g., polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively). Known emulsifying agents include, but are not limited to, lecithin, and acacia. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl-para-hydroxybenzoates, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin. Known thickening agents for oily suspensions include, for example, beeswax, hard paraffin, and cetyl alcohol. 
     Liquid solutions of the active ingredient in aqueous or oily solvents may be prepared in substantially the same manner as liquid suspensions, the primary difference being that the active ingredient is dissolved, rather than suspended in the solvent. As used herein, an “oily” liquid is one which comprises a carbon-containing liquid molecule and which exhibits a less polar character than water. Liquid solutions of the pharmaceutical composition of the invention may comprise each of the components described with regard to liquid suspensions, it being understood that suspending agents will not necessarily aid dissolution of the active ingredient in the solvent. Aqueous solvents include, for example, water, and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as  arachis , olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. 
     Powdered and granular formulations of a pharmaceutical preparation of the invention may be prepared using known methods. Such formulations may be administered directly to a subject, used, for example, to form tablets, to fill capsules, or to prepare an aqueous or oily suspension or solution by addition of an aqueous or oily vehicle thereto. Each of these formulations may further comprise one or more of dispersing or wetting agent, a suspending agent, and a preservative. Additional excipients, such as fillers and sweetening, flavoring, or coloring agents, may also be included in these formulations. 
     A pharmaceutical composition of the invention may also be prepared, packaged, or sold in the form of oil-in-water emulsion or a water-in-oil emulsion. The oily phase may be a vegetable oil such as olive or  arachis  oil, a mineral oil such as liquid paraffin, or a combination of these. Such compositions may further comprise one or more emulsifying agents such as naturally occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soybean or lecithin phosphatide, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. These emulsions may also contain additional ingredients including, for example, sweetening or flavoring agents. 
     Methods for impregnating or coating a material with a chemical composition are known in the art, and include, but are not limited to methods of depositing or binding a chemical composition onto a surface, methods of incorporating a chemical composition into the structure of a material during the synthesis of the material (i.e., such as with a physiologically degradable material), and methods of absorbing an aqueous or oily solution or suspension into an absorbent material, with or without subsequent drying. 
     The regimen of administration may affect what constitutes an effective amount. The therapeutic formulations may be administered to the subject either prior to or after a diagnosis of disease. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation. 
     Administration of the compositions of the present invention to a subject, for example, a mammal, including a human, may be carried out using known procedures, at dosages and for periods of time effective to prevent or treat disease. An effective amount of the therapeutic compound necessary to achieve a therapeutic effect may vary according to factors such as the activity of the particular compound employed; the time of administration; the rate of excretion of the compound; the duration of the treatment; other drugs, compounds or materials used in combination with the compound; the state of the disease or disorder, age, sex, weight, condition, general health and prior medical history of the subject being treated, and like factors well-known in the medical arts. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dose range for a therapeutic compound of the invention is from about 1 and 5,000 mg/kg of body weight/per day. One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective amount of the therapeutic compound without undue experimentation. 
     The compound may be administered to a subject as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. It is understood that the amount of compound dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days. For example, with every other day administration, a 5 mg per day dose may be initiated on Monday with a first subsequent 5 mg per day dose administered on Wednesday, a second subsequent 5 mg per day dose administered on Friday, and so on. The frequency of the dose will be readily apparent to the skilled artisan and will depend upon any number of factors, such as, but not limited to, the type and severity of the disease being treated, the type and age of the animal, etc. 
     Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular subject, composition, and mode of administration, without being toxic to the subject. 
     A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. 
     In particular embodiments, it is especially advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding/formulating such a therapeutic compound for the treatment of a disease in a subject. 
     In one embodiment, the compositions of the invention are administered to the subject in dosages that range from one to five times per day or more. In another embodiment, the compositions of the invention are administered to the subject in range of dosages that include, but are not limited to, once every day, every two, days, every three days to once a week, and once every two weeks. It will be readily apparent to one skilled in the art that the frequency of administration of the various combination compositions of the invention will vary from subject to subject depending on many factors including, but not limited to, age, disease or disorder to be treated, gender, overall health, and other factors. Thus, the invention should not be construed to be limited to any particular dosage regime and the precise dosage and composition to be administered to any subject will be determined by the attending physical taking all other factors about the subject into account. 
     Compounds of the invention for administration may be in the range of from about 1 mg to about 10,000 mg, about 20 mg to about 9,500 mg, about 40 mg to about 9,000 mg, about 75 mg to about 8,500 mg, about 150 mg to about 7,500 mg, about 200 mg to about 7,000 mg, about 3050 mg to about 6,000 mg, about 500 mg to about 5,000 mg, about 750 mg to about 4,000 mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about 20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 50 mg to about 1,000 mg, about 75 mg to about 900 mg, about 100 mg to about 800 mg, about 250 mg to about 750 mg, about 300 mg to about 600 mg, about 400 mg to about 500 mg, and any and all whole or partial increments there between. 
     In some embodiments, the dose of a compound of the invention is from about 1 mg and about 2,500 mg. In some embodiments, a dose of a compound of the invention used in compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, a dose of a second compound (i.e., a drug used for treating the same or another disease as that treated by the compositions of the invention) as described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all whole or partial increments thereof. 
     In one embodiment, the present invention is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound or conjugate of the invention, alone or in combination with a second pharmaceutical agent; and instructions for using the compound or conjugate to treat, prevent, or reduce one or more symptoms of a disease in a subject. 
     The term “container” includes any receptacle for holding the pharmaceutical composition. For example, in one embodiment, the container is the packaging that contains the pharmaceutical composition. In other embodiments, the container is not the packaging that contains the pharmaceutical composition, i.e., the container is a receptacle, such as a box or vial that contains the packaged pharmaceutical composition or unpackaged pharmaceutical composition and the instructions for use of the pharmaceutical composition. Moreover, packaging techniques are well known in the art. It should be understood that the instructions for use of the pharmaceutical composition may be contained on the packaging containing the pharmaceutical composition, and as such the instructions form an increased functional relationship to the packaged product. However, it should be understood that the instructions may contain information pertaining to the compound&#39;s ability to perform its intended function, e.g., treating or preventing a disease in a subject, or delivering an imaging or diagnostic agent to a subject. 
     Routes of administration of any of the compositions of the invention include oral, nasal, parenteral, sublingual, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, and (intra)nasal,), intravesical, intraduodenal, intragastrical, rectal, intra-peritoneal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, or administration. 
     Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions that would be useful in the present invention are not limited to the particular formulations and compositions that are described herein. 
     EXPERIMENTAL EXAMPLES 
     The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein. 
     Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the present invention and practice the claimed methods. The following working examples therefore are not to be construed as limiting in any way the remainder of the disclosure. 
     Example 1: Acetyl-CoA Synthetase Regulates Histone Acetylation and Hippocampal Memory 
     Metabolic production of acetyl coenzyme A (acetyl-CoA) is linked to histone acetylation and gene regulation, but the precise mechanisms of this process are largely unknown. The data presented herein demonstrates that the metabolic enzyme acetyl-CoA synthetase 2 (ACSS2) directly regulates histone acetylation in neurons and spatial memory in mammals. In a neuronal cell culture model, ACSS2 increases in the nuclei of differentiating neurons and localizes to upregulated neuronal genes near sites of elevated histone acetylation. A decrease in ACSS2 lowers nuclear acetyl-CoA levels, histone acetylation, and responsive expression of the cohort of neuronal genes. In adult mice, attenuation of hippocampal ACSS2 expression impairs long-term spatial memory, a cognitive process that relies on histone acetylation. A decrease in ACSS2 in the hippocampus also leads to defective upregulation of memory-related neuronal genes that are pre-bound by ACSS2. These results reveal a connection between cellular metabolism, gene regulation, and neural plasticity and establish a link between acetyl-CoA generation ‘on-site’ at chromatin for histone acetylation and the transcription of key neuronal genes. 
     The observation that ACSS2 is highly expressed in the mouse hippocampus (Lein, E. S. et al., 2007, Nature, 445:168-176) led us to investigate the role of ACSS2 in neuronal histone acetylation and gene expression. These findings support the hypothesis that neuronal ACSS2 has a critical function in linking acetate metabolism to neuronal gene regulation through direct binding of chromatin by ACSS2, and identify a prominent role of this mechanism in hippocampal memory consolidation. 
     The materials and methods employed in these experiments are now described. 
     Mouse Experiments. 
     No Statistical Methods were Used to Predetermine Sample Size; Prior experiments using the relevant behavioral assays with pharmacological or genetic manipulations determined that effects are achieved when group sizes are at least 7-9 animals. The experiments were randomized and the investigators were blinded to allocation during experiments and outcome assessment. 
     Cell Culture 
     CAD cells (Cath.-a-differentiated) were grown in Dulbecco&#39;s modified Eagle&#39;s medium (DMEM):Ham&#39;s F12 (1:1), supplemented with 2 mM glutamine, 1% penicillin/streptomycin, and 10% fetal bovine serum (FBS). To induce neuronal differentiation, sub-confluent CAD cell cultures (50-60%) were transferred to serum-free medium (DMEM:Ham&#39;s F12 (1:1) supplemented with 2 mM glutamine) and maintained in 15-cm 2  culture dishes for 5 days. Upon differentiation, CAD neurons exhibit morphological changes that are characteristic of neurons. For knockdown experiments, CAD cells were infected with lentiviral hairpin constructs (TRC collection) designed against ACL (# TRCN0000055217) or ACSS2 (# TRCN0000076124, # TRCN0000076125) in medium containing 8 mg/mL polybrene and 10% FBS for 24 hours. Cells then underwent selection in culture medium supplemented with 0.5 mg/mL puromycin for 5 days to obtain a stably infected population. Cell treatment with ACSS2i (1-(2,3-di(thiophen-2-yl)quinoxalin-6-yl)-3-(2-methoxyethyl)urea (DMSO)) was carried out for 24 hours at a final concentration of 20 μM (treatment with DMSO alone served as control). 
     RNA-Seq 
     To generate libraries for RNA-seq, poly(A) +  RNA was extracted using the Dynabeads mRNA Direct kit (Ambion) according to the manufacturer&#39;s instructions. RNA-seq libraries for scrambled control (referred to as wild-type), shACL, and shACSS2 were made using a ScriptSeq v2 RNA-seq Library Preparation Kit (Illumina). The quantity and quality of the libraries were assessed by BioAnalyzer (Agilent) and qPCR (Kapa Biosystems). The multiplexed libraries were pooled and sequenced on a single lane on the Illumina NextSeq 500 platform (50 bp, single-end reads). All RNA-seq data were prepared for analysis as follows: NextSeq sequencing data was demultiplexed using bcl2fastq2-v02.14.01.07. Demultiplexed FASTQs were aligned by RNA-STAR 2.3.0.e using the genome index mm10 generated from iGenome UCSC mm10 FASTQ genome sequence. The aligned reads were mapped to genomic features using cufflinks-2.2.1, (-G parameter to quantify only known features), and iGenomes mml0 UCSC genomic transcript loci. The rRNA, mRNA, and tRNA of the mouse genome were downloaded from the goldenPath UCSC FTP and were masked from the transcript quantification. After quantification, all data processing was done using python pandas library v.0.14.0. Differential expression in CAD neurons was defined as the top 10% of genes by fold-change, corresponding roughly to 1.6-fold upregulation or higher. Differential expression in the inhibitor and the hippocampal ACSS2 knockdown in vivo were defined using Cuffdiff. The relationship between CAD cell differentiation and inhibitor function ( FIG. 1G ) was inferred by assessing standardized scores over two RNA-seq replicates each of untreated and ACSS2i-treated cells. These were averaged and genes with |z|&lt;0.5 in either condition were dropped. Scores for remaining genes were plotted in order of increasing CAD differentiation fold-change. The statistical significance of the trend and reproducibility were assessed by taking the top 20% of genes by loss of expression in the knockdown and comparing the expression of these genes in inhibitor-treated cells to a random sample of genes outside this set (Mann-Whitney test). 
     ChIP-Seq 
     CAD cells were fixed in 1% formaldehyde for 10 min and fixation was quenched with the addition of glycine to 125 mM for an additional 5 min. Cells were harvested by scraping from plates, and washed twice in 1×PBS before storage at −80° C. ChIP was performed as previously described (Shah, P. P. et al., 2013, Genes Dev., 27:1787-1799), except that chromatin was sheared to an average size of &lt;500 bp using the Covaris S220 Ultrasonicator. Equal aliquots of sonicated chromatin from undifferentiated and differentiated CAD neurons were used for each immunoprecipitation reaction, and 10% of the amount was saved as input. ACSS2 ChIPs were performed using 2,000 μg extract and 4 μg antibody per sample; all other ChIPs were performed using 500 μg extract and 4 μg antibody per sample. Immunoprecipitation was performed using protein A Dynabeads (Life Technologies). Sequencing libraries were prepared using NEBNext Ultra library preparation procedure, and then assessed for quality and quantity by BioAnalyzer (Agilent) and qPCR (Kapa Biosystems). Sequencing was performed on the Illumina NextSeq 500 platform. All ChIP-seq data were prepared for analysis as follows: NextSeq sequencing data was demultiplexed using bcl2fastq. All reads were aligned to the mm9 or the mml0 reference genome using bowtie2.2.1. One alignment was allowed per read and one mismatch was allowed in the seed region (-N1 -k1). Reads were tabulated in fixed windows or to genes provided in the iGenome mml0 UCSC annotations using featureCounts from the subread 1.4.6 software package. CAD cell ACSS2 ChIP-seq data were normalized to input controls, while all histone acetylation ChIP-seq data were H3-subtracted. The plot in  FIG. 9I  is the result of performing a multiple linear regression to determine the relationship between expression in undifferentiated and differentiated CAD cells (regressors) and enrichment of ACSS2 in differentiated CAD cells (target). The relationship was used to color the negative space in the plot by propensity for ACSS2 binding. For the in vivo ChIP, hippocampal tissue pooled from two animals was finely minced and cross-linked with formaldehyde (1% final concentration) for 15 min at room temperature, followed by glycine quenching for an additional 10 min at 4° C. To create a single-cell suspension, samples were washed once with ice-cold PBS and homogenized by passing through a 22 G needle 10 times. Subsequent steps were performed in the same way as described for the in vitro ChIP. In vivo ChIP peaks (ACSS2 (T), H3K9ac, CBP-GSM1629373, and H3K27ac-GSM1629397) were called using MACS v2.1.0 with the false discover rate (FDR) controlled at 1%. Peak scores were assessed by adjusting to millions of aligned tags and subtracting background. Tracks were similarly normalized and are visualized using the UCSC genome browser with a maximum value windowing function and smoothing at 5 pixels (in vitro ChIP-seq) or using default parameters (in vivo ChIP-seq). The Venn diagram in  FIG. 3B  shows overlap of genes (official gene symbol) that feature ChIP-seq peaks within 1 kb of their nearest TSS. Venn diagrams in  FIG. 3E  and  FIG. 11B  display overlapping target gene sets (RefSeq transcripts) that were assigned to the entire set of ChIP-seq peaks, because CBP binds to few genes within 1 kb of the TSS. The in vivo mRNA expression versus ChIP analysis ( FIG. 3C ) was created by sorting genes on log 2-transformed expression values (DESeq library-size adjusted, replicate-averaged values) in home cage control mice then displaying ChIP-seq AUCs (RPM-adjusted ChIP minus RPM-adjusted background, length adjusted) for each gene for the peak closest to its TSS within a distance of 1 kb (all genes with more distant peaks were rendered as having a score of zero). Gene targets were inferred by the presence of peaks proximal to the TSS (within 1 kb). To identify enriched motifs ( FIG. 3F ), in vivo ACSS2 peaks targeting genes that were upregulated in differentiated CAD cells (without respect for distance to the nearest TSS) were compared to a background set of equal-sized regions selected from gene-rich regions using HOMER (peak sizes were fixed at 300 bp). Discovered motifs were filtered for those present at one-third or more of the targeting peaks and with a tenfold or higher enrichment over the gene-rich background. To assess the overlap of ACSS2 and histone acetylation ( FIG. 7F ,  FIG. 7G ), ACSS2 peaks were filtered to include only those upstream of their nearest target genes. Downstream acetylation was assessed for similarly filtered peaks of H3K9ac, H4K5ac, and H4K12ac from the same cells, as well as cortical H3K27ac. For the in vivo analysis ( FIG. 3B ), gene targets of ACSS2 or H3K9ac peaks within 1 kb of their nearest TSS were examined for overlap. The acetylation pattern due to differentiation at induced or inhibitor-sensitive genes ( FIG. 3D ,  FIG. 7P ) was assessed by taking a 20-kb window around the TSS and measuring the input-adjusted ChIP-seq signal. H3K9ac data were validated ( FIG. 11A ) by comparing to ENCODE&#39;s common-replicate peaks for H3K9ac in mouse forebrain (accession ENCSR369RBO) using CEAS (default parameters, with a 1-10 kb window around the TSS and TES). Additional comparisons were made to H3K9ac (NCBI GEO: GSE82643) and H3K27ac (GSE82428), contrasting to input (GSE82659) to control for sonication efficiency. Cortical H3K27ac (NCBI GEO: GSM1629397) and CBP (GSM1629373) were aligned along with the corresponding input (GSM1629381) using bowtie2 (parameters -k 1 -N 1-local) and peaks were called using MACS2 (input control, FDR controlled at 1%) ( FIG. 3E ,  FIG. 11B ). The combined effect of ACSS2 and histone acetylation targeting on gene expression in vivo ( FIG. 3D ) was demonstrated by box-plotting expression in home cage control mice at genes targeted by ACSS2 by itself, H3K9ac by itself, ACSS2+H3K9ac, or neither. Only genes bound at the promoter (1 kb distance) by ACSS2 were considered. 
     Acetyl-CoA Quantification 
     To extract and quantify acetyl-CoA from differentiated CAD neurons, 4×10 6  cells were washed and incubated in lysis buffer for 30 min (10 mM Tris pH 8, 1 mM KCl, 1.5 mM MgCl 2 , 1 mM DTT). The nuclei were pelleted at 3,000 g for 5 min, and immediately re-suspended in Acetyl CoA Assay Buffer provided in the PicoProbe Acetyl CoA Assay Kit (Abcam, ab87546). The acetyl-CoA assay, including the deproteinization step, was prepared according to the manufacturer&#39;s instructions. The PicoProbe assay was performed in 96-well clear-bottom plates, and the resulting fluorescence was quantified using the Synergy HTX Multi-Mode Microplate Reader (BioTek Instruments). 
     Western Blots 
     Cells were lysed in buffer containing 50 mM Tris pH 8.0, 0.5 mM EDTA, 150 mM NaCl, 1% NP40, 1% SDS, supplemented with protease inhibitor cocktail (Life Technologies, number 78446). For subcellular fractionation experiments, the cells were processed using the subcellular fractionation kit for cultured cells (Thermo Scientific, number 78840) according to the manufacturer&#39;s instructions. Protein concentration was determined by BCA protein assay (Life Technologies, number 23227), and equal amounts of protein were used in co-immunoprecipitation experiments or directly loaded onto polyacrylamide gels. The endogenous co-immunoprecipitation experiments were performed using antibody-conjugated protein A Dynabeads (Life Technologies) in buffer containing: 20 mM Tris, pH 8.0, 137 mM NaCl, 1 mM MgCl 2 , 1 mM CaCl 2 , 1% NP-40, 10% glycerol, with protease and phosphatase inhibitors, and 12.5 U/mL benzonase (Novagen, 70746). Proteins or co-immunoprecipitation eluates were loaded and separated on 4-12% Bis-Tris polyacrylamide gels (NuPAGE). After transfer to nitrocellulose membrane, 3% BSA in TBS supplemented with 0.1% Tween 20 (TBST) was used to block the membrane at room temperature for 1 hour. Primary antibodies were diluted in TBST, and incubated at 4° C. overnight. Primary antibodies are listed below. The membrane was washed three times with TBST, each for 10 min, followed by incubation with HRP-conjugated secondary antibodies at room temperature for 1 hour, in TBST. The membrane was washed again three times, and imaged with a Fujifilm LAS-4000 imager. Original gel blots are provided as  FIGS. 14-16 . 
     Immunofluorescence 
     Cells were fixed in 4% PFA in PBS for 20 min at room temperature. Cells were washed twice with PBS and permeabilized with 0.5% Triton X-100 in PBS for 10 min. After being washed twice, cells were blocked in 10% BSA in PBS for 1 hour at room temperature. Cells were incubated with primary antibodies in 5% BSA in PBS supplemented with 0.1% Tween 20 (PBST) overnight at 4° C. Antibodies are listed below. Then cells were washed four times with PBST, each for 10 min, followed by incubation with fluorophore-conjugated secondary antibody in 5% BSA in PBST for 1 hour at room temperature. F-actin was labelled using Alexa Fluor 488 Phalloidin (Thermo A12379). Cells were then washed three times in PBST, once with PBS, and incubated with 1 μg/mL DAPI for 5 min. The cells were then washed twice with PBS and mounted with ProLong Gold (Invitrogen). The slides were observed and imaged using a Nikon Eclipse microscope. Microscopy settings were unchanged between samples. 
     Antibodies 
     The antibodies used were anti-H3 (Abcam ab1791), anti-H3K9ac (Abcam ab4441), anti-H3K27ac (Abcam ab4729), anti-H3K122ac (Abcam ab33308), anti-H4 (Millipore 05-858), anti-H4K5ac (Millipore 39-584), anti-H4K12ac (Abcam ab1761), anti-ACSS2 (T) (Thermo MA5-145810), anti-ACSS2 (CS) (Cell Signaling 3658), anti-ACL (Proteintech 15421-1-AP), anti-α-tubulin (Sigma T8328), anti-GAPDH (Fitzgerald Industries 10R-G109A), anti-KAT3A/CBP (Abcam ab2832), anti-SNAP25 (Abcam ab5666), anti-synaptophysin (Millipore MAB368), anti-MAP2 C/D (Cell Signaling 8707), anti-NR4A2 (Santa Cruz sc-991) and anti-NeuN (Millipore ABN78). 
     Intracranial Injection of Viral Vector 
     Adult mice (8+ weeks of age) were anaesthetized with isoflurane gas (1-5% to maintain surgical plane) and placed in a sterile field within a stereotaxic device. Animals received an injection of bupivacaine (2.5 mg/kg) for local anaesthesia before the skin was disinfected with betadine solution and the skull exposed with a short incision using sterile surgical equipment. Artificial tears were applied to eyes to ensure sufficient lubrication. A small hole (about 0.5 mm) was drilled in the skull over the target area using a stereotax and a stereotactic drill. A micro-syringe filled with viral vector was inserted into the dorsal hippocampus and slowly removed following injection (AP, −2.0 mm; DV, −1.4 mm; ML, +1.5 mm from bregma). ACSS2 knockdown vector, AAV2/9.U6.shACSS2.CMV.EGFP; eGFP control vector, AAV2/9.CMV.EGFP.polyA. All animals received a single dose of subcutaneous meloxicam (5 mg/kg) as analgesia at induction and one dose per day for two days postoperatively as needed. 
     Object Location Memory Task 
     The object location memory procedure is used to test spatial memory. The procedure consists of a training phase and a testing phase. Prior to training, each mouse was handled for 3 min a day for 3 days. On the training day, mice are placed in an arena (approx. 1 square foot) containing three different objects. The objects used were a glass bottle, a metal tower (h×w×l, 5×2×2 inches), and a plastic cylinder. Mice were habituated to an empty arena with a black and white striped spatial cue on the wall, followed by object exposure in three 6-min trials with an interval of 3 min. The arena and objects were cleaned with 70% EtOH between trials. To diminish biases, the memory test was performed on control and knockdown mice on the same day in the same arena, using every combination of object location (n=10 mice per study group). After 24 hours, the individual mice were placed back in the arena used in the testing phase. For testing, one of the objects was moved to different location in the arena. Mice were allowed to explore freely for 5 min. Each session was recorded using a video camera and time spent exploring (approaches and sniffing) each object was assessed offline. All animals were randomized and preassigned to arena and object the day before testing to ensure that every treatment group explored every object configuration. 
     Contextual Fear Conditioning 
     The mouse was placed in the conditioning chamber (Med Associates) for 5 min before the onset of the unconditioned stimulus (US), a 1.5-mA continuous foot shock. A mild 2-sec, 1.5-mA foot shock is used as an aversive stimulus; this does not injure the mice but provides the transient, yet startling and aversive, stimulus that is necessary for conditioning. After an additional 30 sec in the chamber, the mouse was returned to its home cage. Twenty-four hours later, the mouse was tested for a freezing response to the chamber (contextual) where training occurred. Time spent freezing in the chamber (motionless except for respiratory movements) was assessed for 5 consecutive minutes. 
     Data Availability 
     The ChIP-seq and RNA-seq data have been made available at the Gene Expression Omnibus (GEO) repository under the SuperSeries accession code GSE76854. 
     The results of the experiments are now described. 
     ACSS2 Regulates Neuronal Gene Expression 
     The function of ACSS2 in neurons was investigated using the Cath.-a-differentiated (CAD) cell line derived from mouse catecholaminergic cells. Upon serum deprivation, CAD cells differentiate to form neuronal processes and become excitable, similar to functional neurons (Qi, Y. et al., 1997, J. Neurosci., 17:1217-1225). Immunofluorescence showed that endogenous ACSS2 was primarily cytoplasmic in undifferentiated CAD cells ( FIG. 1A ), but shifted primarily to the nucleus upon differentiation ( FIG. 1B ,  FIG. 5A ). Whole-cell and nuclear levels of ACSS2 increased upon differentiation of CAD cells into neurons, whereas cytoplasmic ACL expression remained constant ( FIG. 1C ). In primary hippocampal and cortical neurons from mouse brain, even 14 days after isolation, ACSS2 was chiefly nuclear and ACL was primarily cytoplasmic ( FIG. 5C ,  FIG. 5D ,  FIG. 5E ,  FIG. 5F ). It was concluded that ACSS2, unlike ACL, is localized to nuclei during neuronal differentiation. 
     The role of ACSS2 in upregulation of canonical neuron-specific protein markers in differentiated CAD neurons was investigated. Pre-differentiation knockdown of ACSS2 reduced differentiation-linked expression of nuclear NeuN, activity-regulated Nr4a2, and the cytoplasmic markers synaptophysin, Map2 and Snap25, without an associated decrease in ACL ( FIG. 5G ), indicating that ACSS2 has a key role in neuronal differentiation. 
     Transcriptome analysis by mRNA sequencing (mRNA-seq) upon CAD neuronal differentiation identified 894 upregulated genes ( FIG. 7A ,  FIG. 7B ,  FIG. 7C ; Table 1). Gene ontology analysis revealed that these differentiation-linked genes were neuron-specific; gene ontology terms included neuron differentiation, synaptic transmission, ion transport, and neuron projection morphogenesis ( FIG. 7E ). A protein interaction framework that produced a neuronal network centered on activity-dependent signaling and synaptic plasticity was developed: calmodulin 1 (Calm1), glutamate ionotropic receptor NMDA type subunit 1 (Grin1), and inositol 1,4,5-trisphosphate receptor type 1 (Itpr1) ( FIG. 7D ). Calm1 mediates the control of neuronal proteins by Ca 2+  during synaptic plasticity, including Ca 2+ /calmodulin-dependent protein kinase II (CaMKII). Such Ca 2+  signaling is regulated by Grin1, an NMDA receptor subtype of glutamate-gated ion channels, and also by the ion channel Itpr1, which mobilizes intracellular Ca 2+  stores, an important process in activity-dependent signaling that underlies synaptic plasticity during learning. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 A list of genes upregulated 1.6-fold or higher upon CAD neuronal differentiation, 
               
               
                 corresponding to the top 10% of upregulated genes by fold-change diff vs undiff. 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Fold- 
                   
                 Fold- 
                   
                 Fold- 
               
               
                 Gene 
                 change 
                 Gene 
                 change 
                 Gene 
                 change 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Gm1821 
                 175.6579238 
                 Cd3eap 
                 2.170133884 
                 Eme2 
                 1.823970526 
               
               
                 Bc1 
                 18.89222241 
                 B3galt6 
                 2.169981093 
                 Abcb10 
                 1.823927603 
               
               
                 Chgb 
                 17.14357199 
                 Pxylp1 
                 2.166764625 
                 Coa3 
                 1.822527659 
               
               
                 Gm15127 
                 15.51766099 
                 Plxncl 
                 2.160531802 
                 Dnajb12 
                 1.82210436 
               
               
                 Rnaset2b 
                 12.75361823 
                 Camk1d 
                 2.158710733 
                 Mad111 
                 1.821858573 
               
               
                 Tcte3 
                 9.952430312 
                 Myt11 
                 2.155525888 
                 Nagpa 
                 1.821421835 
               
               
                 Slc7a14 
                 8.513220194 
                 2310040G24Rik 
                 2.154691887 
                 Bag5 
                 1.820414157 
               
               
                 Syt4 
                 7.711544335 
                 Nagk 
                 2.15371853 
                 Scg5 
                 1.820055865 
               
               
                 Chga 
                 6.960140476 
                 Zfp646 
                 2.153449442 
                 Cyb561d2 
                 1.819353638 
               
               
                 Tcte3 
                 6.787934727 
                 Clcn5 
                 2.153194258 
                 Slc7a1 
                 1.818877568 
               
               
                 St18 
                 6.154430487 
                 D430020J02Rik 
                 2.152743565 
                 Dmrtc1c1 
                 1.81721542 
               
               
                 Lrrn3 
                 5.678039797 
                 Ptprr 
                 2.152060138 
                 Dusp16 
                 1.816831604 
               
               
                 Nefh 
                 5.663029385 
                 Neur13 
                 2.151224453 
                 Tfpi 
                 1.815801097 
               
               
                 Sv2c 
                 5.604691346 
                 Rtn3 
                 2.149814381 
                 Smarcad1 
                 1.815095829 
               
               
                 St8sia3 
                 5.516463854 
                 Coro2a 
                 2.148771712 
                 Kidins220 
                 1.815049028 
               
               
                 Tfrc 
                 5.330857918 
                 Syne2 
                 2.146800723 
                 Tex261 
                 1.81297524 
               
               
                 C7 
                 5.088574779 
                 Kcnq2 
                 2.146167325 
                 Itga6 
                 1.812508999 
               
               
                 Ina 
                 5.036193249 
                 Cd151 
                 2.145739959 
                 Jph3 
                 1.812352364 
               
               
                 6030419C18Rik 
                 5.000233138 
                 Trmt5 
                 2.145499 
                 Gm14139 
                 1.810377573 
               
               
                 Lix1 
                 4.924417409 
                 Opcml 
                 2.142896705 
                 Asphd1 
                 1.810011841 
               
               
                 F2rl2 
                 4.868341078 
                 Ptpn13 
                 2.142246807 
                 Smarce1 
                 1.807946088 
               
               
                 Bex2 
                 4.733589022 
                 Vegfc 
                 2.140464179 
                 Rrp7a 
                 1.807062004 
               
               
                 Srp54b 
                 4.688634976 
                 Gm13154 
                 2.140261646 
                 Myh3 
                 1.806367241 
               
               
                 Chrna3 
                 4.611736933 
                 Gm9833 
                 2.140118183 
                 Lin28b 
                 1.805854386 
               
               
                 Bend7 
                 4.585426322 
                 Tmem158 
                 2.138968007 
                 Nup85 
                 1.804665858 
               
               
                 Cp1x1 
                 4.444224944 
                 Fscn1 
                 2.137050753 
                 Scrt1 
                 1.803337363 
               
               
                 Scg2 
                 4.356337617 
                 Rundc3a 
                 2.13434778 
                 Atf2 
                 1.80172488 
               
               
                 Syp 
                 4.327573468 
                 Spint1 
                 2.133736983 
                 Serpina1d 
                 1.801250346 
               
               
                 Itga1 
                 4.278299803 
                 Gng4 
                 2.129537408 
                 Dbn1 
                 1.80099271 
               
               
                 Hmgb3 
                 4.1862105 
                 Ms1312 
                 2.128923538 
                 Ankrd49 
                 1.80079943 
               
               
                 Gpr22 
                 4.1674996 
                 Dnajc6 
                 2.128248925 
                 Celf5 
                 1.799789076 
               
               
                 Gm6644 
                 4.106399081 
                 Arxes2 
                 2.124711148 
                 Pam 
                 1.799761546 
               
               
                 Ngfrap1 
                 4.090845268 
                 Hexim1 
                 2.124625821 
                 Zbtb10 
                 1.799707942 
               
               
                 11-Mar 
                 4.082591127 
                 Cstf2 
                 2.123424958 
                 Magea8 
                 1.799361507 
               
               
                 Lrp11 
                 4.046236641 
                 Apol10b 
                 2.123145491 
                 Kcnb1 
                 1.798921382 
               
               
                 Syne1 
                 3.991300602 
                 Garem 
                 2.121754047 
                 Agpat9 
                 1.797386751 
               
               
                 Grb14 
                 3.945690545 
                 Mmp24 
                 2.116735802 
                 Dfna5 
                 1.796745137 
               
               
                 Myb 
                 3.90824467 
                 Siah1b 
                 2.113360374 
                 Gnao1 
                 1.795924972 
               
               
                 S1c26a4 
                 3.870330962 
                 9330182L06Rik 
                 2.113260274 
                 Onecut2 
                 1.795915729 
               
               
                 Gap43 
                 3.845523746 
                 Klf11 
                 2.111304581 
                 Nop2 
                 1.795704394 
               
               
                 Tubb3 
                 3.839461152 
                 Nrxn2 
                 2.108670556 
                 Cxx1c 
                 1.795202843 
               
               
                 Agtr1a 
                 3.787641152 
                 Ttyh3 
                 2.106168371 
                 Tbkbp1 
                 1.795132889 
               
               
                 Cobll1 
                 3.776724233 
                 Tmem63c 
                 2.104274635 
                 Lamp1 
                 1.793429305 
               
               
                 Manea1 
                 3.74209634 
                 Ppfia2 
                 2.104105162 
                 Uros 
                 1.792154061 
               
               
                 Ablim1 
                 3.696297385 
                 Calm1 
                 2.101694438 
                 Olfml3 
                 1.791288679 
               
               
                 Pyg1 
                 3.66431466 
                 Act16b 
                 2.100910703 
                 Immt 
                 1.79036382 
               
               
                 Ogfod3 
                 3.623862166 
                 Stmn3 
                 2.100007128 
                 N1rp4c 
                 1.789285052 
               
               
                 Zdbf2 
                 3.595183027 
                 Efna5 
                 2.098388029 
                 Tmod2 
                 1.789055398 
               
               
                 Tcte3 
                 3.563270202 
                 Rhbdd2 
                 2.097503081 
                 Gsg2 
                 1.788260344 
               
               
                 Zcchc12 
                 3.555338395 
                 5730409E04Rik 
                 2.096839768 
                 Brpf3 
                 1.788146652 
               
               
                 Insm1 
                 3.51414208 
                 A330076H08Rik 
                 2.09651456 
                 Psme3 
                 1.787606977 
               
               
                 Rtl1 
                 3.445197877 
                 Celf4 
                 2.093422103 
                 B230217O12Rik 
                 1.787068322 
               
               
                 Bhlhb9 
                 3.427368454 
                 Chst11 
                 2.091694127 
                 Dhrs7b 
                 1.786833183 
               
               
                 Rpl26 
                 3.403059425 
                 Trmt6 
                 2.091481094 
                 Mfn2 
                 1.786829419 
               
               
                 Fmn12 
                 3.390037903 
                 Tle4 
                 2.091242414 
                 Clu 
                 1.786603419 
               
               
                 Diras2 
                 3.344496731 
                 Mid2 
                 2.090168909 
                 Mdn1 
                 1.784727192 
               
               
                 Syt7 
                 3.305395031 
                 Lrif1 
                 2.083927045 
                 Crlf1 
                 1.782967462 
               
               
                 Nrp1 
                 3.254754196 
                 Aplp2 
                 2.080116197 
                 Dusp1 
                 1.78268785 
               
               
                 Crmp1 
                 3.252645244 
                 C1ql1 
                 2.076261716 
                 I123a 
                 1.782195797 
               
               
                 Trp53i11 
                 3.241772978 
                 Ets2 
                 2.076132879 
                 Zfp60 
                 1.780426997 
               
               
                 Elfn2 
                 3.238729769 
                 Gde1 
                 2.07447251 
                 Mcts1 
                 1.779483364 
               
               
                 Acyp2 
                 3.234343027 
                 Mras 
                 2.074382859 
                 Shq1 
                 1.77924886 
               
               
                 Snap91 
                 3.230834555 
                 Nt5c2 
                 2.073514495 
                 Grhl2 
                 1.7791477 
               
               
                 Cpt1a 
                 3.229336895 
                 Acsl5 
                 2.072620341 
                 Trappc2 
                 1.778257911 
               
               
                 Gpatch4 
                 3.222172569 
                 Chrm4 
                 2.072223233 
                 Cd9 
                 1.777982221 
               
               
                 Tceal8 
                 3.217666679 
                 Rom1 
                 2.07128601 
                 Ppp2r3d 
                 1.777014646 
               
               
                 Bex1 
                 3.188074487 
                 Kcnn2 
                 2.068794964 
                 3300005D01Rik 
                 1.776683404 
               
               
                 4933432K03Rik 
                 3.183007869 
                 Dusp22 
                 2.0667913 
                 Trappc21 
                 1.775573052 
               
               
                 Ubxn8 
                 3.180294016 
                 Serinc1 
                 2.065964409 
                 Atp6v1h 
                 1.774994978 
               
               
                 Nme1 
                 3.179596125 
                 Grin1 
                 2.063481712 
                 BC005624 
                 1.774906486 
               
               
                 Gm3448 
                 3.176047756 
                 Ccdc40 
                 2.061884422 
                 Nipa2 
                 1.773384434 
               
               
                 Eid2 
                 3.163737628 
                 Imp4 
                 2.059494693 
                 Armcx4 
                 1.772224038 
               
               
                 Rgs7 
                 3.140441667 
                 Sync 
                 2.058561575 
                 Sptbn1 
                 1.771756217 
               
               
                 Sez612 
                 3.138031104 
                 Itpr1 
                 2.058179448 
                 Acs16 
                 1.771288672 
               
               
                 Nudt1 
                 3.131824273 
                 Dlg2 
                 2.057014709 
                 Exoc8 
                 1.770448693 
               
               
                 Ttc8 
                 3.127218279 
                 Laptm4a 
                 2.055520048 
                 Tmx2 
                 1.770238878 
               
               
                 Ccdc64 
                 3.113624626 
                 Tmx4 
                 2.054452 
                 Apmap 
                 1.769567355 
               
               
                 Soga3 
                 3.106573768 
                 Ccdc711 
                 2.053367712 
                 Dpcd 
                 1.76832124 
               
               
                 Dynlt1f 
                 3.075927839 
                 Tex13 
                 2.052406599 
                 Lactb 
                 1.767193082 
               
               
                 Kcnab1 
                 3.075671599 
                 Ccdc106 
                 2.049420856 
                 BC048403 
                 1.766450957 
               
               
                 Myt1 
                 3.070224982 
                 Ofd1 
                 2.049383373 
                 Mcm10 
                 1.766240198 
               
               
                 Rpp25 
                 3.069442261 
                 Atrnl1 
                 2.048809708 
                 Hipk3 
                 1.766048616 
               
               
                 Capn6 
                 3.06860951 
                 Fbxl16 
                 2.047921237 
                 Gnaz 
                 1.765435041 
               
               
                 Rps6ka6 
                 3.051147613 
                 Mia2 
                 2.04777355 
                 Tnik 
                 1.765229777 
               
               
                 Moap1 
                 3.050801905 
                 Hs3st2 
                 2.047395008 
                 Ywhah 
                 1.765199109 
               
               
                 Sult4a1 
                 3.050032271 
                 Scamp5 
                 2.04672811 
                 Gm4944 
                 1.764668491 
               
               
                 Thsd7a 
                 3.042155585 
                 Cd248 
                 2.042574215 
                 Pik3r1 
                 1.764390036 
               
               
                 Syt1 
                 3.035831847 
                 Myo1b 
                 2.042361194 
                 Mcm2 
                 1.763793482 
               
               
                 Dppa2 
                 3.025823893 
                 Kbtbd8 
                 2.039844229 
                 Pgbd5 
                 1.763785993 
               
               
                 Sobp 
                 3.016460969 
                 Dscaml1 
                 2.035957043 
                 Ahnak 
                 1.762687909 
               
               
                 Glcci1 
                 3.005342056 
                 Arsb 
                 2.034765917 
                 Tubg1 
                 1.762408455 
               
               
                 Lin28a 
                 2.983923999 
                 Glipr1 
                 2.027994233 
                 Upf3b 
                 1.761898167 
               
               
                 Usp51 
                 2.975360783 
                 Smarcc1 
                 2.026414744 
                 Vav2 
                 1.761024602 
               
               
                 Eno1b 
                 2.97263557 
                 Dscc1 
                 2.026254958 
                 Cmpk2 
                 1.760528848 
               
               
                 Ubqln2 
                 2.956614456 
                 Atp6v1c1 
                 2.024670168 
                 Flrt3 
                 1.759648874 
               
               
                 Arrdc3 
                 2.955284729 
                 Tub 
                 2.024311372 
                 2900009J06Rik 
                 1.7594006 
               
               
                 Isg15 
                 2.933241785 
                 Ctxn1 
                 2.021976572 
                 Apon 
                 1.758839365 
               
               
                 Pde4dip 
                 2.906021486 
                 Nudc 
                 2.021960351 
                 A330040F15Rik 
                 1.756818568 
               
               
                 Shisa3 
                 2.881691875 
                 Eif3b 
                 2.020017253 
                 Dpysl3 
                 1.756580466 
               
               
                 Dync1h1 
                 2.873386193 
                 Napb 
                 2.017401664 
                 Wdr6 
                 1.756565798 
               
               
                 Cpeb2 
                 2.854892675 
                 Cdc6 
                 2.016230466 
                 Zfp345 
                 1.756041804 
               
               
                 Syt6 
                 2.84753409 
                 Gprasp1 
                 2.015850089 
                 Gm5801 
                 1.755666178 
               
               
                 Clvs1 
                 2.822978842 
                 Nfasc 
                 2.015721892 
                 Vipas39 
                 1.755094357 
               
               
                 Kcnk3 
                 2.817797535 
                 Nxf3 
                 2.014974572 
                 Pih1d2 
                 1.753692794 
               
               
                 Gm5868 
                 2.812232401 
                 Zhx1 
                 2.012540964 
                 Abcc3 
                 1.752792921 
               
               
                 Arxes1 
                 2.811860001 
                 B3galt1 
                 2.010886035 
                 Mki67 
                 1.751856644 
               
               
                 Stox2 
                 2.780799101 
                 Gba 
                 2.006904267 
                 5033406O09Rik 
                 1.751135413 
               
               
                 Nmnat2 
                 2.779691083 
                 Mafb 
                 2.005616568 
                 Sema6d 
                 1.750218169 
               
               
                 Basp1 
                 2.776507236 
                 Icam2 
                 2.005565245 
                 Mapkap1 
                 1.748722597 
               
               
                 Nxt2 
                 2.765596658 
                 Csrnp3 
                 2.004149481 
                 Rmdn3 
                 1.748669909 
               
               
                 Slco3a1 
                 2.761274885 
                 G530011O06Rik 
                 2.003834543 
                 Fut11 
                 1.747886156 
               
               
                 9330159F19Rik 
                 2.759766806 
                 Ispd 
                 2.00357033 
                 Rufy3 
                 1.747069267 
               
               
                 Bscl2 
                 2.752674021 
                 Trmt61a 
                 1.999997328 
                 Fam21 
                 1.746467678 
               
               
                 Zbtb1 
                 2.750343915 
                 Ccpg1 
                 1.998744702 
                 Angptl7 
                 1.74568611 
               
               
                 Slc25a20 
                 2.732767312 
                 Sox11 
                 1.997046172 
                 Ptgr2 
                 1.744488542 
               
               
                 Lrr1 
                 2.724323049 
                 Upf3a 
                 1.992047853 
                 Parp8 
                 1.741931597 
               
               
                 H2afy2 
                 2.714935834 
                 Dst 
                 1.991416813 
                 Rims4 
                 1.74001918 
               
               
                 Ccdc101 
                 2.688893985 
                 Slc35g2 
                 1.991032922 
                 Mex3b 
                 1.739919034 
               
               
                 Prps2 
                 2.688066633 
                 Dusp10 
                 1.990659306 
                 Hmha1 
                 1.739634453 
               
               
                 Slit2 
                 2.680804093 
                 Zfp672 
                 1.989832275 
                 Sh3kbp1 
                 1.739580052 
               
               
                 Fam73a 
                 2.680489857 
                 Tmem56 
                 1.98880423 
                 Cbll1 
                 1.738638987 
               
               
                 Spock2 
                 2.668728711 
                 Xkr5 
                 1.988679099 
                 Tspan14 
                 1.738560062 
               
               
                 Id4 
                 2.663294856 
                 Chrnb2 
                 1.98824031 
                 Fam136a 
                 1.736003183 
               
               
                 Elavl3 
                 2.661848056 
                 Cisd2 
                 1.988057414 
                 Ampd3 
                 1.735262497 
               
               
                 Gprin1 
                 2.659163353 
                 Alg2 
                 1.984414687 
                 Cdk5r2 
                 1.733761696 
               
               
                 Zfp941 
                 2.653595168 
                 Abcb6 
                 1.979958221 
                 Phox2b 
                 1.7326033 
               
               
                 Arhgef28 
                 2.652052288 
                 Arpp21 
                 1.979082597 
                 Utp20 
                 1.732574299 
               
               
                 Pcdhac2 
                 2.645536483 
                 Eml6 
                 1.97800923 
                 Pigt 
                 1.732092573 
               
               
                 Cited2 
                 2.635020506 
                 Pvrl1 
                 1.977863127 
                 Rab11b 
                 1.73169755 
               
               
                 Gabrb3 
                 2.630267867 
                 Ccne2 
                 1.976366942 
                 Bdh1 
                 1.731670577 
               
               
                 Mmd 
                 2.630198806 
                 Hist2h3c1 
                 1.975040976 
                 Lrrc2 
                 1.731121108 
               
               
                 Trpc7 
                 2.62761764 
                 Hist2h3c1 
                 1.975040506 
                 Mettl24 
                 1.730670243 
               
               
                 Tmem164 
                 2.622650194 
                 Hist2h3c1 
                 1.97439534 
                 9130023H24Rik 
                 1.730622105 
               
               
                 Astn1 
                 2.619175631 
                 Hist2h3c1 
                 1.974394871 
                 Slc35f3 
                 1.730212207 
               
               
                 Lmo2 
                 2.613888268 
                 Pelo 
                 1.973649509 
                 Cyb5r4 
                 1.728975276 
               
               
                 Kpnb1 
                 2.612082447 
                 Pmpca 
                 1.971199074 
                 4933409K07Rik 
                 1.728946459 
               
               
                 Ywhag 
                 2.611362467 
                 D14Ertd670e 
                 1.966194908 
                 Fbll1 
                 1.728705387 
               
               
                 Pcdha9 
                 2.609047858 
                 Acap3 
                 1.964633963 
                 Iba57 
                 1.728152072 
               
               
                 4930412O13Rik 
                 2.59954161 
                 Naa40 
                 1.96318494 
                 Abhd16a 
                 1.724790788 
               
               
                 Sez6 
                 2.594545132 
                 Odc1 
                 1.962296385 
                 C230052I12Rik 
                 1.724519072 
               
               
                 Mthfd1 
                 2.594534783 
                 Nsf 
                 1.962079336 
                 Cep290 
                 1.723738934 
               
               
                 Cygb 
                 2.594202918 
                 Ric3 
                 1.957504324 
                 Disp2 
                 1.721375855 
               
               
                 Mapk8ip2 
                 2.586130338 
                 Cyb561 
                 1.956983455 
                 Ccdc86 
                 1.7206313 
               
               
                 Rims2 
                 2.582500994 
                 Prpf31 
                 1.950424024 
                 Slc24a2 
                 1.718144872 
               
               
                 Zfp105 
                 2.57834921 
                 Atp1a3 
                 1.949704023 
                 Naaa 
                 1.716002932 
               
               
                 Rab3c 
                 2.56905558 
                 Asic1 
                 1.949220769 
                 Myef2 
                 1.71556509 
               
               
                 Tcerg11 
                 2.560508109 
                 Prkar2a 
                 1.947217244 
                 Marcks11 
                 1.714941564 
               
               
                 Lgr5 
                 2.559906391 
                 Gm10516 
                 1.94574251 
                 As1 
                 1.712968358 
               
               
                 Ank2 
                 2.557088209 
                 Gdap111 
                 1.945511639 
                 Pmm2 
                 1.712746409 
               
               
                 AA414768 
                 2.556426952 
                 Shisa4 
                 1.944591761 
                 Rtn4rl1 
                 1.712680069 
               
               
                 Fen1 
                 2.552951046 
                 Jph1 
                 1.943999212 
                 Rltpr 
                 1.712475563 
               
               
                 Atp2b3 
                 2.552525124 
                 2410076I21Rik 
                 1.942836136 
                 Rbm3 
                 1.711669169 
               
               
                 Plcxd3 
                 2.547915161 
                 Zfhx4 
                 1.942750871 
                 Ccdc137 
                 1.711399973 
               
               
                 Map1b 
                 2.546432223 
                 B230216N24Rik 
                 1.942106125 
                 Tubb2a 
                 1.710451318 
               
               
                 Cnga3 
                 2.54463692 
                 Pomt2 
                 1.941883342 
                 Cers6 
                 1.71012081 
               
               
                 Prima1 
                 2.540015798 
                 Ak1 
                 1.940188779 
                 Rc3h2 
                 1.70930883 
               
               
                 Tceal1 
                 2.536928591 
                 Hmgn2 
                 1.939312621 
                 Wfikkn2 
                 1.709206434 
               
               
                 Ptprn 
                 2.530300281 
                 Crb2 
                 1.938608613 
                 D6Wsu163e 
                 1.708994488 
               
               
                 Gm14124 
                 2.528627525 
                 Sncb 
                 1.938186969 
                 Fam171a2 
                 1.708684719 
               
               
                 Nsg1 
                 2.527832329 
                 Mcm7 
                 1.938141062 
                 Slc8a3 
                 1.708154517 
               
               
                 Peg13 
                 2.526460165 
                 Nav3 
                 1.937091743 
                 Tubb2b 
                 1.707297937 
               
               
                 Chrna7 
                 2.521656827 
                 Celsr3 
                 1.937008133 
                 Psmc1 
                 1.706891691 
               
               
                 Frmpd1 
                 2.513212694 
                 Nomo1 
                 1.936027801 
                 Chd4 
                 1.706378216 
               
               
                 Scg3 
                 2.507811675 
                 Ppip5k2 
                 1.935223601 
                 Zdhhc24 
                 1.7057495 
               
               
                 Map7d2 
                 2.497575084 
                 Fads6 
                 1.934377481 
                 Kifap3 
                 1.705719758 
               
               
                 Cltb 
                 2.497372532 
                 Zdhhc2 
                 1.933684461 
                 Cadm1 
                 1.705546607 
               
               
                 Podx1 
                 2.496009524 
                 Prep 
                 1.932286093 
                 Fdxacb1 
                 1.705207276 
               
               
                 Gpr68 
                 2.495164795 
                 Pfdn2 
                 1.929900656 
                 Dcun1d2 
                 1.704817808 
               
               
                 Syn1 
                 2.492408986 
                 Aox4 
                 1.927911942 
                 Gpr19 
                 1.704379411 
               
               
                 Peg3os 
                 2.485925677 
                 2010320M18Rik 
                 1.926539948 
                 Atl1 
                 1.704121204 
               
               
                 Stac 
                 2.485547115 
                 Rrp12 
                 1.926113939 
                 Zfp37 
                 1.702563195 
               
               
                 1700008J07Rik 
                 2.484043438 
                 Tnfsf13b 
                 1.92507446 
                 Galc 
                 1.70230816 
               
               
                 Tmtc1 
                 2.482449248 
                 Pdss1 
                 1.922389127 
                 K1h17 
                 1.701799864 
               
               
                 Akap12 
                 2.481981855 
                 Ddx25 
                 1.92174518 
                 Coro1c 
                 1.701663394 
               
               
                 Gm13889 
                 2.473754193 
                 Ppat 
                 1.920879419 
                 N4bp2 
                 1.70137242 
               
               
                 Aig1 
                 2.471662617 
                 Rad51 
                 1.920722212 
                 Cyth3 
                 1.700454504 
               
               
                 Nap115 
                 2.469349003 
                 A430035B10Rik 
                 1.920377197 
                 Gria2 
                 1.699598134 
               
               
                 Ptprn2 
                 2.467641397 
                 Rimkla 
                 1.920131006 
                 Rnf103 
                 1.699576595 
               
               
                 Il1b 
                 2.467520516 
                 Fkrp 
                 1.918435117 
                 Rybp 
                 1.699136669 
               
               
                 Fstl4 
                 2.461319176 
                 Gng2 
                 1.914720195 
                 Mgat2 
                 1.698388185 
               
               
                 Gdap1 
                 2.459324502 
                 Zbtb6 
                 1.913889619 
                 Cacng3 
                 1.698216152 
               
               
                 Hist1h2bc 
                 2.458592095 
                 Cacna1b 
                 1.91297449 
                 Pou4f2 
                 1.698190252 
               
               
                 Rgs4 
                 2.456820051 
                 Sfxn1 
                 1.912605475 
                 Slc25a16 
                 1.697489578 
               
               
                 Rtn2 
                 2.456045107 
                 Ddx24 
                 1.910286019 
                 Tmem5 
                 1.697359997 
               
               
                 Qsox2 
                 2.443479002 
                 Dapk1 
                 1.909272865 
                 Ankrd45 
                 1.69578789 
               
               
                 Slc10a4 
                 2.443317686 
                 Mfsd1 
                 1.90858969 
                 Med1 
                 1.695016446 
               
               
                 Npcd 
                 2.442844829 
                 Kcnma1 
                 1.906252243 
                 Scn3a 
                 1.693976545 
               
               
                 Psmc5 
                 2.43852755 
                 Gnl31 
                 1.905404777 
                 Knop1 
                 1.693141571 
               
               
                 Spire2 
                 2.43817828 
                 Dapp1 
                 1.905280655 
                 Letm2 
                 1.692996703 
               
               
                 Ppp2r2b 
                 2.436623889 
                 Fig4 
                 1.904074862 
                 Pcna 
                 1.692987099 
               
               
                 Rab27a 
                 2.431185467 
                 Akr1c13 
                 1.903698431 
                 Srm 
                 1.692546245 
               
               
                 Tcte3 
                 2.430285346 
                 Eefla2 
                 1.902891155 
                 H3f3b 
                 1.692540018 
               
               
                 Unc79 
                 2.42896397 
                 Epb4.113 
                 1.902831138 
                 Rhov 
                 1.692258256 
               
               
                 Rere 
                 2.420946998 
                 Ovca2 
                 1.901183855 
                 Cinp 
                 1.692148625 
               
               
                 Nkrf 
                 2.418071053 
                 Ssh2 
                 1.900453866 
                 Nek3 
                 1.690663139 
               
               
                 Tshz2 
                 2.41642778 
                 Prnp 
                 1.899301236 
                 Nol9 
                 1.690524211 
               
               
                 Lhfpl4 
                 2.410185371 
                 Mrps2 
                 1.899212746 
                 Gm8801 
                 1.690275316 
               
               
                 Isl1 
                 2.409248695 
                 Tmtc4 
                 1.898938507 
                 Setbp1 
                 1.690128797 
               
               
                 Srrm3 
                 2.403958838 
                 Taf3 
                 1.897880465 
                 Mtss1 
                 1.689835118 
               
               
                 Slc29a4 
                 2.396866695 
                 Ddx6 
                 1.897322975 
                 Champ1 
                 1.689414384 
               
               
                 Hsp90aa1 
                 2.394800411 
                 Bex4 
                 1.896950555 
                 Secisbp2 
                 1.688464915 
               
               
                 Galnt3 
                 2.387693032 
                 Rpp38 
                 1.896133169 
                 Fus 
                 1.687897894 
               
               
                 Fam46a 
                 2.386772798 
                 Mmp15 
                 1.896113253 
                 Zfp936 
                 1.687393579 
               
               
                 Akap6 
                 2.385151202 
                 Dnajb9 
                 1.896089418 
                 Umps 
                 1.687287148 
               
               
                 Unc5a 
                 2.385077118 
                 6430548M08Rik 
                 1.893669196 
                 Erlin2 
                 1.686403998 
               
               
                 Spa17 
                 2.381932467 
                 Gusb 
                 1.893273972 
                 Ncs1 
                 1.68635252 
               
               
                 Armcx1 
                 2.381036751 
                 Ptprj 
                 1.892506369 
                 Trim67 
                 1.685742571 
               
               
                 Rundc3b 
                 2.377774829 
                 Tro 
                 1.890403492 
                 Ckmt1 
                 1.685073896 
               
               
                 Kif5c 
                 2.374822358 
                 Nos1 
                 1.889456863 
                 Arhgap39 
                 1.684943724 
               
               
                 Polr1a 
                 2.37006057 
                 Ulbp1 
                 1.889367146 
                 Syn2 
                 1.684313334 
               
               
                 Dhrs4 
                 2.365368437 
                 Atp11c 
                 1.888641934 
                 Zdhhc6 
                 1.683756464 
               
               
                 Smarca2 
                 2.365086661 
                 H60b 
                 1.886035789 
                 Morc2a 
                 1.683594931 
               
               
                 Sdc3 
                 2.364573718 
                 Lrrc4b 
                 1.885291767 
                 5830444B04Rik 
                 1.683180352 
               
               
                 Impact 
                 2.358940049 
                 Runx3 
                 1.885204635 
                 Nol10 
                 1.683138348 
               
               
                 Syt14 
                 2.3576276 
                 Tmed1 
                 1.884337236 
                 Cacng4 
                 1.683101168 
               
               
                 Akr1c12 
                 2.35170279 
                 Cpsf2 
                 1.883974673 
                 Dnajc7 
                 1.682611037 
               
               
                 Rsph1 
                 2.351486036 
                 Tmem57 
                 1.882975491 
                 Pdzd2 
                 1.682587463 
               
               
                 Pmp22 
                 2.350035976 
                 2310033P09Rik 
                 1.882250099 
                 Pak6 
                 1.682545384 
               
               
                 Wbp5 
                 2.346786732 
                 Rabl2 
                 1.882158882 
                 Ptplad1 
                 1.679458325 
               
               
                 Prkca 
                 2.344303821 
                 Pitpna 
                 1.881932453 
                 Lipo1 
                 1.679354818 
               
               
                 Rnf113a2 
                 2.34299707 
                 Cdc123 
                 1.881701502 
                 Clcn4-2 
                 1.679179827 
               
               
                 Rtn4 
                 2.341355269 
                 Psmd1 
                 1.881543038 
                 2210016F16Rik 
                 1.678795515 
               
               
                 Dusp3 
                 2.336914633 
                 Gpatch3 
                 1.881016859 
                 Fgd6 
                 1.677856945 
               
               
                 Vgf 
                 2.333281785 
                 Gabarap11 
                 1.879939416 
                 Bend3 
                 1.676649255 
               
               
                 Pja1 
                 2.332545789 
                 Lhfpl5 
                 1.878997447 
                 Pomgnt2 
                 1.676509151 
               
               
                 Lrch2 
                 2.327787274 
                 Zfp386 
                 1.87838411 
                 Stk32c 
                 1.676286848 
               
               
                 Xkr7 
                 2.323195789 
                 Ppp1r13b 
                 1.877446456 
                 Kif3b 
                 1.675950473 
               
               
                 Msrb2 
                 2.321774461 
                 Akap1 
                 1.876716578 
                 Tug1 
                 1.675280054 
               
               
                 Resp18 
                 2.320094281 
                 Clec21 
                 1.876692332 
                 Dcakd 
                 1.675043345 
               
               
                 Ftsj3 
                 2.319925457 
                 Fam3c 
                 1.87512747 
                 Hnrnpu 
                 1.674090524 
               
               
                 Rrm2 
                 2.318816915 
                 Gla 
                 1.874879429 
                 6530402F18Rik 
                 1.673841373 
               
               
                 3-Sep 
                 2.317901491 
                 Fancm 
                 1.872317503 
                 AI507597 
                 1.67311856 
               
               
                 Dusp26 
                 2.315217495 
                 Isoc1 
                 1.87105305 
                 Lrrc3 
                 1.673042164 
               
               
                 Atp6v0a1 
                 2.314441552 
                 Palld 
                 1.870884616 
                 Osbpl10 
                 1.673029921 
               
               
                 Mfsd11 
                 2.306371316 
                 Jph4 
                 1.869670861 
                 6330409D20Rik 
                 1.672704362 
               
               
                 Sstr2 
                 2.305792842 
                 Cnrip1 
                 1.869633707 
                 Fam222a 
                 1.672294928 
               
               
                 Grip1 
                 2.303010673 
                 Dars 
                 1.868613783 
                 4930474H20Rik 
                 1.671611988 
               
               
                 Hnrnph2 
                 2.301444559 
                 Ddx21 
                 1.867198424 
                 Mast1 
                 1.671433547 
               
               
                 Nipal2 
                 2.298812802 
                 Hcn4 
                 1.865788038 
                 Trp53bp1 
                 1.671138201 
               
               
                 Ston2 
                 2.295055047 
                 Fam43a 
                 1.864011078 
                 Baz2b 
                 1.67075683 
               
               
                 Vps53 
                 2.289633653 
                 Vwa5b1 
                 1.863969349 
                 Gm1140 
                 1.669835745 
               
               
                 Gipc2 
                 2.289236832 
                 Arhgdig 
                 1.862404417 
                 Dcaf5 
                 1.669626713 
               
               
                 Lhx5 
                 2.287726502 
                 Trmt2b 
                 1.862289161 
                 Iscu 
                 1.669593253 
               
               
                 Dclk1 
                 2.284738254 
                 Meis3 
                 1.861831561 
                 Thumpd1 
                 1.669127846 
               
               
                 Snap25 
                 2.282197664 
                 Gemin6 
                 1.861333656 
                 Fam171a1 
                 1.668887754 
               
               
                 Bcap29 
                 2.282122018 
                 Gm5512 
                 1.861333198 
                 Tmed10 
                 1.668819621 
               
               
                 Syt11 
                 2.281088449 
                 Ppp1r10 
                 1.861029556 
                 Pi4ka 
                 1.66870687 
               
               
                 Tanc2 
                 2.278569711 
                 Sox4 
                 1.86083136 
                 Ccar1 
                 1.668501964 
               
               
                 Prkar1b 
                 2.277882216 
                 Rnf149 
                 1.859016185 
                 Usp31 
                 1.668019551 
               
               
                 Ttll7 
                 2.276918784 
                 Gm10451 
                 1.858282261 
                 D3Ertd751e 
                 1.667496489 
               
               
                 Ppp2r2c 
                 2.276658072 
                 Stmn4 
                 1.856774327 
                 Pdap1 
                 1.667488844 
               
               
                 Armcx2 
                 2.268644676 
                 Unc13a 
                 1.85671842 
                 Leo1 
                 1.667471581 
               
               
                 Wdr35 
                 2.265314142 
                 Aven 
                 1.853946127 
                 Dut 
                 1.666931066 
               
               
                 Lgals3bp 
                 2.265115028 
                 Stau2 
                 1.85156307 
                 Slc39a6 
                 1.665602627 
               
               
                 Eif4e3 
                 2.264987807 
                 Gamt 
                 1.851189856 
                 Grwd1 
                 1.665476983 
               
               
                 Map2 
                 2.264668982 
                 1700052K11Rik 
                 1.850918723 
                 Bop1 
                 1.665146841 
               
               
                 Rgl1 
                 2.264094249 
                 Wasf1 
                 1.85029807 
                 Baz1a 
                 1.664496169 
               
               
                 Tmx1 
                 2.260736628 
                 Klc1 
                 1.84994572 
                 Ncapg2 
                 1.664133825 
               
               
                 Rhox5 
                 2.260459126 
                 Mapt 
                 1.849853184 
                 Ptpn1 
                 1.663963745 
               
               
                 2010204K13Rik 
                 2.256574437 
                 Sdc1 
                 1.849803409 
                 Dhx9 
                 1.661797652 
               
               
                 Hcn2 
                 2.253035203 
                 Zfp428 
                 1.849336547 
                 A330050F15Rik 
                 1.661663284 
               
               
                 Gm7120 
                 2.249641702 
                 Pigw 
                 1.848547815 
                 Tmem131 
                 1.661052736 
               
               
                 Dynll2 
                 2.247730864 
                 Garnl3 
                 1.848497313 
                 Exosc9 
                 1.661033821 
               
               
                 Nrip1 
                 2.236297154 
                 Mrpl20 
                 1.846810463 
                 Agfg1 
                 1.661002122 
               
               
                 Panx1 
                 2.235271455 
                 Smarca4 
                 1.846436526 
                 Ahsa1 
                 1.660962974 
               
               
                 Slc16a6 
                 2.233992923 
                 Efhc2 
                 1.846083405 
                 Lgmn 
                 1.660328735 
               
               
                 Mt3 
                 2.2331597 
                 Pcdhgb1 
                 1.845930958 
                 Pacsin1 
                 1.660310806 
               
               
                 Celf3 
                 2.233013864 
                 Mycbpap 
                 1.845365822 
                 Gm6787 
                 1.659350326 
               
               
                 Dnajc11 
                 2.219751251 
                 Wdfy3 
                 1.845013089 
                 Gstm7 
                 1.658970823 
               
               
                 Pkia 
                 2.219406778 
                 Nceh1 
                 1.84493714 
                 Fmnl1 
                 1.658654117 
               
               
                 Commd9 
                 2.219393748 
                 Fam134b 
                 1.844817405 
                 Rnf112 
                 1.657680566 
               
               
                 4930526I15Rik 
                 2.218644904 
                 Pstpip1 
                 1.844427603 
                 Ptcd3 
                 1.657152744 
               
               
                 Se1113 
                 2.217920637 
                 Elav14 
                 1.843902306 
                 Hhex 
                 1.656754506 
               
               
                 4930550L24Rik 
                 2.210086462 
                 Tuba8 
                 1.843254105 
                 Ufm1 
                 1.656676294 
               
               
                 Foxp1 
                 2.208954244 
                 Rapgef2 
                 1.842648206 
                 Smarcd1 
                 1.656415594 
               
               
                 LOC100503496 
                 2.208155647 
                 Mcm6 
                 1.842531069 
                 Dyrk3 
                 1.656199454 
               
               
                 Fam111a 
                 2.206805703 
                 Dmrtc1c2 
                 1.842393283 
                 Bid 
                 1.656100175 
               
               
                 Tmie 
                 2.205900235 
                 Sgsh 
                 1.842238312 
                 Dync2li1 
                 1.656027888 
               
               
                 Unc80 
                 2.203512779 
                 Dph6 
                 1.84038813 
                 Zfp11 
                 1.654777396 
               
               
                 Tha1 
                 2.19982455 
                 Meis2 
                 1.839755932 
                 Smoc1 
                 1.654537223 
               
               
                 Prps113 
                 2.199715007 
                 0610040B10Rik 
                 1.839304618 
                 Nfe212 
                 1.654021972 
               
               
                 Stmn2 
                 2.199601839 
                 Mllt11 
                 1.838902842 
                 Atl3 
                 1.652987408 
               
               
                 Ap4s1 
                 2.198216557 
                 Pnmal1 
                 1.838598117 
                 Ubl3 
                 1.652814032 
               
               
                 Smpd3 
                 2.197857241 
                 BC039966 
                 1.836049546 
                 Tmem33 
                 1.651936589 
               
               
                 Smarca1 
                 2.197803474 
                 Nin 
                 1.83534734 
                 Slc4a3 
                 1.651755144 
               
               
                 Tspan7 
                 2.196692952 
                 Tmem74 
                 1.834876695 
                 Tenm3 
                 1.650505091 
               
               
                 Pctp 
                 2.195340118 
                 Ccdc92 
                 1.833747074 
                 Ap1s2 
                 1.650381217 
               
               
                 Mapk10 
                 2.193496445 
                 Plxna4 
                 1.832242996 
                 Vma21 
                 1.650080813 
               
               
                 Ap3b2 
                 2.189548862 
                 Nkain1 
                 1.82998511 
                 Fbxl19 
                 1.648861973 
               
               
                 Gpm6b 
                 2.188414017 
                 Timm8a1 
                 1.82935184 
                 Hist3h2a 
                 1.648031868 
               
               
                 Sumf1 
                 2.184806171 
                 Srrm4 
                 1.828687427 
                 Gm15663 
                 1.647544686 
               
               
                 Kif1b 
                 2.181037857 
                 Atp6v0d1 
                 1.828497312 
                 Apbb1 
                 1.64735093 
               
               
                 Tmem42 
                 2.179489111 
                 Ube4a 
                 1.828181709 
                 Psmc3 
                 1.647297037 
               
               
                 Apex1 
                 2.175711582 
                 Eif4g2 
                 1.828117898 
                 Naip2 
                 1.646832206 
               
               
                 Mblac2 
                 2.174911584 
                 Coq7 
                 1.827888701 
                 Fbxo47 
                 1.646673454 
               
               
                 Cask 
                 2.173206274 
                 Tusc3 
                 1.826480198 
                 Gemin4 
                 1.644644567 
               
               
                 Brsk2 
                 2.172446145 
                 Gtf2a1 
                 1.826336279 
                 Naip6 
                 1.643373664 
               
               
                 Cdkl2 
                 2.171509052 
                 Nolc1 
                 1.82454386 
                 Hk1 
                 1.643066226 
               
               
                   
               
            
           
         
       
     
     The metabolite acetyl-CoA is required by HAT enzymes for histone acetylation. To investigate histone acetylation during differentiation of CAD cells into neurons, chromatin immunoprecipitation with high-throughput DNA sequencing (ChIP-seq) was performed for histone H3 lysine 9 acetylation (H3K9ac), H4K5ac, and H4K12ac (see Methods). All marks were enriched upon differentiation at upregulated neuronal genes (for example, at nudix-type motif 1 (Nudt1)) ( FIG. 6F ). Overall, the 894 upregulated neuronal genes displayed higher acetylation than all other genes ( FIG. 6G ). 
     The levels of ACSS2 or ACL were reduced using short hairpin RNAs (shRNA) before cell differentiation and RNA-seq ( FIG. 6H ,  FIG. 6I ,  FIG. 6J ,  FIG. 6K ). The induction of neuronal genes was lost in the ACSS2 knockdown cells (Pearson r=0.15;  FIG. 1D ,  FIG. 1E ), whereas the same genes retained a strong correlation in transcriptional fold-change in ACL knockdown cells (Pearson r=0.53;  FIG. 5L ). The top 10% upregulated genes in differentiated wild-type cells were stratified into quintiles ( FIG. 1I ), and it was found that ACSS2 knockdown strongly lowered upregulation across all quintiles ( FIG. 11 : green bars; P=7.2×10 −252 , Wilcoxon rank-sum test). The ACL-knockdown cells showed the same upward trend as wild-type cells, in contrast to the severe defect in ACSS2-knockdown cells ( FIG. 6M ; P=1.1×10 −25 , Wilcoxon rank-sum test). Notably, ACL-knockdown cells showed lower global transcript levels (P=1.91×10 −7 , Mann-Whitney U-test), unlike ACSS2-knockdown cells, which showed a less severe genome-wide defect ( FIG. 6N ; P=0.04, Mann-Whitney U-test). ACL thus has a broad but non-specific effect on gene expression, whereas ACSS2 is required for upregulation of the neuronal gene expression program upon differentiation of CAD cells into neurons. 
     Further, it was tested whether ACSS2 catalytic activity is required for the neuronal gene expression program using a small-molecule specific inhibitor of ACSS2 (ACSS2i) (Comerford, S. A. et al., 2014, Cell, 159:1591-1602). RNA-seq showed a reduction in differentiation-induced genes ( FIG. 1G ), and the genes whose expression was affected by ACSS2-knockdown were also highly sensitive to the ACSS2i ( FIG. 6O , P=1.62×10 6 ). 
     Recruitment of ACSS2 to Chromatin 
     The direct association of ACSS2 with chromatin was investigated using ChIP-seq of differentiated CAD cells (see Methods). Two ACSS2 antibodies showed a high correlation both for model-based analysis of ChIP-seq (MACS) overlapping peaks (Spearman r=0.82;  FIG. 7A ), and for global enrichment over 1-kb genomic windows (Spearman r=0.73;  FIG. 7B ). Binding of ACSS2 correlated with increases in histone H3K9ac, H4K5ac, and H4K12ac in differentiated relative to undifferentiated CAD cells, for instance at the promoters of Nudt1 and Tab2 (TAK1-binding protein 2;  FIG. 7C ,  FIG. 7D ). Both genes have been linked to neurodegenerative disorders; the Nudt1 hydrolase oxidizes purine nucleoside triphosphates to prevent RNA incorporation, and Tab2 regulates signal transduction pathways in neurons (Sardi, S. P. et al., 2006, Cell, 127:185-197). Gene ontology analysis showed that genes proximal to ACSS2 peaks were linked to neuronal differentiation ( FIG. 7E ). Hence, chromatin-associated, neuronal gene promoter-proximal ACSS2 may provide a local source of acetyl-CoA to HAT enzymes. 
     ACSS2 binding relative to histone acetylation was examined, and it was found that 80% of ACSS2 peaks upstream of the nearest target gene overlapped an acetylation peak or had an acetylation peak downstream towards the targeted transcription start site (TSS;  FIG. 7F ,  FIG. 7G ). A substantial number (13-15% of all ACSS2 peaks genome-wide) directly overlapped peaks of H3 and H4 acetylation ( FIG. 7H ). In addition, the height of ACSS2 peaks correlated overall with intersected histone acetylation peaks ( FIG. 7I ,  FIG. 7J ,  FIG. 7K ). This peak height correlation suggests that H4 acetylation might be most responsive to ACSS2-derived acetyl-CoA, in particular H4K12ac, a mark that has been linked to defective memory formation during aging (Peleg, S. et al., 2010, Science, 328:753-756). In general, the most enriched ACSS2 peaks displayed the strongest histone acetylation enrichment ( FIG. 7L ,  FIG. 7M ,  FIG. 7N ). 
     Putative recruitment of ACSS2 by transcription factor binding was investigated using de novo motif discovery over ACSS2 ChIP-seq peaks, which revealed binding sequences predicted for neuronal transcription factors. The most enriched motif was Yin Yang1 (YY1) ( FIG. 7O ; P=1×10 −599 ), which recruits the two acetyl-CoA-dependent HAT enzymes CREB-binding protein (CBP) and E1A binding protein (p300) (17), consistent with the idea that ACSS2 fuels nearby catalytic HAT activity. 
     Initial peak analysis did not identify all peaks of ACSS2 or acetylation ( FIG. 8A ,  FIG. 8B ), so gene body enrichment was analyzed and there were additional prominent examples, such as Camk2a ( FIG. 2A ), which encodes the CaMKII alpha chain that is required for hippocampal long-term potentiation (LTP) and spatial learning. ACSS2 and acetylation co-occupancy profiles were similar at Camk2a and Nudt1 ( FIG. 7C ). Meta-gene analysis indicated that the top 5% of ACSS2-enriched genes had levels of acetylation up to threefold higher than the mean across all genes ( FIG. 9A ,  FIG. 9B ,  FIG. 9C ,  FIG. 9D ), and genes with the greatest fold-change in differential ACSS2 binding had the highest histone acetylation levels ( FIG. 2C ;  FIG. 9E ,  FIG. 9F ,  FIG. 9G ,  FIG. 9H ). Gene ontology term enrichment showed that the top ACSS2-bound and acetylated genes were neuron-specific ( FIG. 2B ). 
     At all induced genes, ACSS2 binding was concomitant with increased histone acetylation ( FIG. 7P ), and the 299 genes that showed reduced expression upon ACSS2i treatment were those with the greatest differentiation-linked increases in histone acetylation ( FIG. 2D ). In total, about 9% of genes previously linked to neuronal differentiation (ND genes, AmiGO annotation set of 1,315 genes) were induced in differentiated CAD cells, and these induced genes were exceptionally sensitive to ACSS2i treatment ( FIG. 2E , ‘Induced’). Moreover, although the entire ND gene class did not change expression in differentiated CAD cells, their expression was markedly reduced by ACSS2i treatment ( FIG. 2E , ‘ND genes’). The interaction between differentiation-linked gene expression changes and ACSS2 recruitment to chromatin was visualized using multiple linear regression analysis, and a remarkable relationship between higher ACSS2 enrichment (red) and increased gene expression was found ( FIG. 9I ). Overall, the CAD cell genomic data demonstrate dynamic ACSS2 enrichment in differentiated neurons linked to increased histone acetylation and involvement in transcriptional upregulation of neuronal genes. 
     ACSS2 Functions in Neuronal Histone Acetylation 
     Nuclear acetyl-CoA levels was measured in ACSS2 knockdown cells ( FIG. 2F ; mean Δ=−0.19±0.03, P=0.003) and in cells treated with ACSS2i ( FIG. 2F ; mean Δ=−0.25±0.05, P=0.006) and it was found that levels of acetyl-CoA were similarly decreased. This finding supports the theory that ACSS2 enzymatic activity supplies nuclear acetyl-CoA. Global histone acetylation levels of transcription-linked H3K27ac and H3K9ac were reduced in ACSS2 knockdown cells ( FIG. 2G ,  FIG. 10A ), and these marks are key substrates of the transcriptional coactivators CBP and p300 with roles in hippocampal LTP and long-term memory (18). ACSS2 co-immunoprecipitated with acetylated chromatin, specifically H3K9ac, H3K27ac, and H4K12ac ( FIG. 10B ), and also with CBP ( FIG. 2H ), suggesting that ACSS2 binds chromatin at transcriptionally active genes to increase histone acetylation during memory formation in vivo (Wood, M. A. et al., 2005, Learn. Mem., 12:111-119; Korzus, E. et al., 2004, Neuron, 42:961-972; Vecsey, C. G. et al., 2007, J. Neurosci., 27:6128-6140). 
     ACSS2 was examined in primary mouse hippocampal neurons, given their capacity for depolarization and expression of key memory-related neuronal genes. ACSS2 was localized to the nucleus ( FIG. 2I ), and ACSS2i treatment reduced neuronal marker expression and histone acetylation without lowering ACSS2 levels ( FIG. 2J ,  FIG. 10C ). Expression of ACL did not change ( FIG. 2J ), indicating that ACL is less important than ACSS2 in the regulation of histone acetylation in hippocampal neurons. 
     Chromatin association of ACSS2 and H3K9ac was assessed in vivo using ChIP-seq in mouse hippocampus. The hippocampal H3K9ac mapping strongly correlated with ENCODE mouse forebrain H3K9ac ChIP-seq (Spearman coefficient of multiple correlation R=0.67), with similar peak distribution ( FIG. 11A ). Hippocampal ACSS2 and H3K9ac corresponded genome-wide and over three canonical neuronal genes involved in memory (Arc, Egr2 and Nr2f2;  FIG. 3A ,  FIG. 3B ). In addition, ACSS2 promoter binding and H3K9ac correlated with RNA-seq in the hippocampus ( FIG. 3C ). A small number of genes were found that were ACSS2-bound but not H3K9ac enriched that were silent, similar to genes not enriched for ACSS2 or H3K9ac ( FIG. 3D ). By contrast, genes enriched for H3K9ac were actively transcribed, but genes enriched for both ACSS2 and H3K9ac were most highly expressed ( FIG. 3D ). 
     Physical association of ACSS2 and CBP in differentiated CAD cells ( FIG. 2H ) correlated with gene colocalization of ACSS2 and CBP in the hippocampus, together with H3K27ac (from public mouse cortex CBP and H3K27ac ChIP-seq data; ACSS2:CBP overlap P=3.23×10 −16  by hypergeometric test). Overall, 57% of ACSS2-associated genes were co-targeted by H3K27ac ( FIG. 3E ), and ACSS2 and CBP co-targeted genes were enriched for gene ontology terms involved in synaptic membrane potential ( FIG. 11B ,  FIG. 11C ). Motif analysis at hippocampal ACSS2 peaks show that Nrf1—a transcription factor that regulates neurite growth-predicted binding at 45% of ACSS2 sites ( FIG. 3F ), evoking an ACSS2-CBP recruitment mechanism. Moreover, ACSS2i-sensitive genes (50%: 145 of 289) had proximal ACSS2 within 10 kb of the TSS (hypergeometric analysis, P=7.6986×10 8 ), consistent with a direct role for chromatin-bound ACSS2 in transcription. 
     ACSS2 Regulates Long-Term Memory Storage 
     Hippocampus-dependent spatial memory occurs through activity-dependent changes in gene expression that are coordinated, in part, through epigenetic modifications, specifically histone acetylation (Barrett, R. M. et al., 2011, Neuropsychopharmacology, 36:1545-1556; Graff, J. et al., 2012, Nat. Commun., 3:991). ACSS2 is expressed throughout the hippocampus (Lein, E. S. et al., 2007, Nature, 445:168-176; Ariyannur, P. S. et al., 2010, J. Comp. Neurol., 518:2952-2977) ( FIG. 12A ), and thus could mediate histone acetylation to upregulate neuronal gene expression during memory consolidation (Barrett, R. M. et al., 2011, Neuropsychopharmacology, 36:1545-1556; Maren, S. et al., 2000, Behav. Brain Res., 110:97-108). To investigate the role of ACSS2 in the adult hippocampus, ACSS2 expression was attenuated in the dorsal hippocampus by shRNA knockdown using a viral vector ( FIG. 4A ,  FIG. 4B ). Compared to control-injected mice, ACSS2 knockdown mice showed similar levels of locomotion, coordination, body weight, and anxiety-related thigmotaxis during open field exploration (Stanford, S. C., 2007, J. Psychopharmacol., 21:134-135) ( FIG. 12B ,  FIG. 12C ,  FIG. 12D ; not significant, n=10 per group); therefore, ACSS2 knockdown did not cause gross behavioral alterations. 
     To assess hippocampus-dependent spatial memory, an object-location memory paradigm was used (Balderas, I. et al., 2008, Learn. Mem., 15:618-624). Animals explored three different objects during training, and long-term memory was tested by re-exposure 24 hours later with one object moved to a different location ( FIG. 4A , right). In training, control and knockdown mice showed no difference in exploration ( FIG. 4C , left). During memory retrieval, control mice showed increased exploration of the object that had been moved ( FIG. 4C ). By contrast, ACSS2 knockdown mice were impaired in spatial object memory ( FIG. 4C  and  FIG. 12E , mean Δ=−5.01±1.21; P=8.3×10 5 ), and displayed a lower discrimination index ( FIG. 4D ; % ΔDI=−29.5±11.4; P=0.02). ACSS2 knockdown mice showed reduced total object exploration during the test ( FIG. 4C ), suggesting diminished novelty associated with intact recognition of the objects from the training session (mean Δ=−6.13±2.15; P=0.02, n=10 per group). 
     Previous studies have shown that long-term contextual fear memory is mediated by the ventral hippocampus when manipulations of the dorsal hippocampus occur before training (Rogers, J. L. et al., 2006, Neurobiol. Learn. Mem., 86:72-81). Therefore, as a control experiment, mice injected with the ACSS2 knockdown shRNA or eGFP in the dorsal hippocampus were subjected to a contextual fear conditioning paradigm. During the 24-hour test session, there was no significant difference in the amount of freezing behavior between ACSS2-knockdown and eGFP mice ( FIG. 12F ,  FIG. 12G ) suggesting that the ventral hippocampus successfully mediated context-shock association. Overall, it was concluded that ACSS2 has a critical role in dorsal hippocampus-mediated long-term spatial memory. 
     ACSS2 Regulates Upregulation of Immediate Early Genes 
     Long-term spatial memory requires a rapid increase in histone acetylation and immediate early gene transcription to occur in a sensitive time window to enable memory consolidation (Barrett, R. M. et al., 2011, Neuropsychopharmacology, 36:1545-1556; Peixoto, L. L. et al., 2015, BMC Genomics, 16:S5); during memory retrieval, gene transcription also occurs for memory reconsolidation, and this can be prevented by inhibiting mRNA synthesis during the sensitive post-retrieval period (Mamiya, N. et al., 2009, J. Neurosci., 29:402-413). It was tested whether ACSS2 was involved in dynamic gene upregulation for hippocampal memory consolidation and reconsolidation by performing mRNA-seq on the dorsal hippocampus. Global gene expression changes were first identified that were induced by spatial object training, which has not previously been investigated genome-wide. Dorsal hippocampi from control and shACSS2-knockdown mice were removed during the sensitive period of memory consolidation following spatial object training. To control for circadian oscillation, injected animals were included that had been handled but not trained. 
     Genes that were differentially expressed following training were identified by transcriptome comparison of trained control-injected mice to untrained circadian control-injected mice using Cuffdiff. A small number of genes were induced immediately after training, including immediate early genes such as Egr2, Fos, Nr2f2, Sgk1 and Arc ( FIG. 4E ). Importantly, baseline expression of these memory-associated genes in untrained control and ACSS2-knockdown mice was remarkably similar ( FIG. 12H ). By contrast, dynamic upregulation of these immediate early genes by training was greatly reduced by ACSS2 knockdown ( FIG. 4E ). 
     It was further tested whether ACSS2 also regulates immediate-early gene expression in the dorsal hippocampus during memory reconsolidation. Therefore, the hippocampal mRNA-seq analysis was focused on previously identified and validated genes that become upregulated following memory retrieval (Peixoto, L. L. et al., 2015, BMC Genomics, 16:S5; Poplawski, S. G. et al., 2014, Neurobiol. Learn. Mem., 116:90-95). Retrieval-associated induction of immediate early genes during the sensitive reconsolidation period was blocked by ACSS2 knockdown, whereas retrieval-linked downregulation during the same period was not ( FIG. 13A ,  FIG. 13B ,  FIG. 13C ,  FIG. 13D ,  FIG. 13E ,  FIG. 13F ). 
     Summary 
     Metabolic state can regulate chromatin structure, in particular by inducing histone modifications (Gut, P. et al., 2013, Nature, 502:489-498). Here, a connection is established between cellular metabolism and neuronal plasticity, and a neuronal function of ACSS2 as a chromatin-bound transcriptional coactivator that stimulates histone acetylation and gene expression is revealed. 
     Acetyl-CoA metabolism is cell- and tissue-specific, and is frequently disregulated in malignant transformation (Comerford, S. A. et al., 2014, Cell, 159:1591-1602; Mashimo, T. et al., 2014, Cell, 159:1603-1614). In adipose cells, ACSS2 partially localizes to the nucleus and contributes to histone acetylation under conditions of low glucose (Wellen, K. E. et al., 2009, Science, 324:1076-1080; Gao, X. et al., 2016, Nat. Commun., 7:11960), but the principal metabolic determinant of histone acetylation is ACL9. By contrast, it is shown herein that post-mitotic neurons rely on chromatin-recruited ACSS2 to supply acetyl-CoA for histone acetylation. Notably, fasting lowers acetyl-CoA and protein acetylation in most tissues, but acetyl-CoA levels remain unchanged in the brain (Marifio, G. et al., 2014, Mol. Cell, 53:710-725), indicating that neuronal ACSS2 has an important role in the fasted state when acetyl-CoA production by citrate-dependent ACL is reduced. 
     Optimal acetyltransferase activity requires an increased local acetyl-CoA to CoA ratio, which determines the catalytic activity and substrate specificity of HAT enzymes (Cai, L., et al., 2011, Mol. Cell, 42:426-437; Wellen, K. E. et al., 2009, Science, 324:1076-1080; Takahashi, H. et al., 2006, Mol. Cell, 23:207-217). This finding suggests that histone acetylation can be controlled by changing levels of nuclear acetyl-CoA. Thus, chromatin-bound ACSS2 could provide acetyl-CoA to fuel HAT activity locally, instantaneously recycling CoA and could also recapture acetate from deacetylation reactions. The data presented herein demonstrates specific chromatin binding by ACSS2 at neuronal genes and link localization to upregulation of histone acetylation and gene transcription in spatial memory ( FIG. 4F ), which requires increased histone acetylation (Graff, J. et al., 2013, Nat. Rev. Neurosci., 14:97-111; Graff, J. et al., 2012, Nat. Commun., 3:991). A crucial role for ACSS2 in upregulation of immediate early genes with key functions in neuronal plasticity and memory is demonstrated, leading to a critical role for this molecule in long-term memory consolidation. 
     Epigenetic mechanisms continue to be revealed as important regulators of neural function and behavior, and have been implicated in neuropsychiatric diseases, including anxiety disorders and depression (Kandel, E. R. et al., 2014, Cell, 157:163-186; Graiff, J. et al., 2013, Nat. Rev. Neurosci., 14:97-111; Walker, D. M. et al., 2015, Curr. Opin. Neurobiol., 30:112-121). In establishing ACSS2 as a key regulator at the interface of metabolic environment and neuronal chromatin, this example provides a previously unrecognized enzymatic target for the development of therapies to treat neurological and cognitive disorders. 
     Example 2: ACSS2 and Alcohol 
     The physiological sources of acetate include 1) acetylated proteins; 2) bacterial fermentation in colon; and 3) ingested ethanol. However, it is unknown if alcohol is destined for neuronal chromatin ( FIG. 17 ). 
     To study the effects of acute alcohol administration, mice are injected intraperitoneally with EtOH- 13 C 2 , to mimic “binge” drinking. Mass spectrometry is used to determine the acetylation of liver and brain histones. Heavy C acetate is incorporated into liver and brain histones by 1 hour post IP injection ( FIG. 18 ). 
     Next, it was determined if ACSS2 is required for heavy C acetate incorporation into hippocampus acetylated histones. ACSS2 KD in dorsal HPC leads to A loss of incorporation into dorsal HPC (4 h). The radar plot displays the relative abundance of the heavy isotope (as compared to the light one). The heavy isotope is higher in the knock-down as compared to wild type only in hippocampus ventral and in muscle. In hippocampus dorsal it remains at the same low levels as in WT mice (injected with unlabeled EtOH). ( FIGS. 19-20 ). 
     Example 3: Liver Alcohol Metabolism Directly Fuels Histone Acetylation in the Brain 
     In the adult brain, epigenetic control of gene expression has important roles in the processing of neural activity. Emerging evidence suggests that epigenetic regulation is dependent on metabolic state, implicating specific metabolic factors in neural functions that drive behavior. In neurons, histone acetylation is dependent on the metabolite acetyl-CoA that is produced from acetate by chromatin-bound ACSS2 (Mews et al., Nature, 2017, 546:381-386). Here, using in vivo stable isotope labeling, it is shown that liver alcohol metabolism rapidly fuels histone acetylation in the brain by direct deposition of alcohol-derived acetyl groups onto histones in an ACSS2-dependent manner. A similar induction was also observed with heavy labeled acetate injection in vivo. In addition, injection of labeled alcohol into a pregnant mouse results in incorporation of labeled acetyl groups into the brains of the gestating fetuses. In isolated primary hippocampal neurons in vitro, extracellular acetate induced learning and memory-related transcriptional programs that were sensitive to ACSS2 inhibition were discovered. These findings establish a novel and direct link between hepatic alcohol metabolism and neuronal histone acetylation, providing the first evidence for dynamic signaling from liver metabolism directly to epigenetic regulation in neurons. 
     Existing research into the neurobiology of alcohol addiction has focused on limbic reward circuitry, changes in neurotransmission, and intracellular neuronal signaling cascades. However, the exact mechanisms by which alcohol exerts its psychotropic effects remain incompletely understood. While alcohol directly interacts with neuronal channel proteins, this pathway cannot explain many of the effects of alcohol intoxication on brain function (Ron et al., Nat. Publ. Gr., 2016, 17:576-591). Indeed, recent work converges on the notion that dysregulated gene expression is a key molecular mechanism of alcohol action on target tissues (Ron et al, Nat. Publ. Gr., 2016, 17:576-591; Zakhari, Alcohol Res., 2013, 35:6-16). In the brain, epigenetic regulation of gene expression enables integration of neural activity to continuously adapt circuit connectivity and has critical importance for expression of appropriate or—in the case of alcohol addiction—inappropriate behaviors (Ron, Nat. Publ. Gr., 2016, 17:576-591; Mews et al., Prog. Brain Res., 2017, 235:19-63; Robinson et al., Nat. Rev. Neurosci., 2011, 12:623-637; Egervari et al., Neurosci. Biobehav. Rev., 2018, 85:117-125). Emerging evidence suggests that such epigenetic regulation is dependent on metabolic state. Indeed, it was recently shown that histone acetylation controlling hippocampal memory formation is reliant on the metabolite acetyl-CoA, produced from acetate by chromatin-bound ACSS2 (Mews et al., Nature, 2017, 546:381-386). Notably, a major physiological source of acetate is via breakdown of alcohol in the liver, leading to rapidly increasing blood acetate (Sarkola et al., Alcohol. Clin. Exp. Res., 2002, 26:239-245). Neuronal histone acetylation may thus be under the influence of extracellular levels of acetate (Soliman et al., Mol. Cell. Biochem., 2011, 352:173-180). However, it is unknown whether hepatic alcohol metabolism directly affects histone acetylation in the brain. 
     The methods and materials are now discussed. 
     Histone Extraction 
     The cells were incubated in nuclear isolation buffer (NIB) (15 mM Tris-HCl, 15 mM NaCl, 60 mM KCl, 5 mM MgCl 2 , 1 mM CaCl 2 ), 250 mM sucrose, pH 7.5, and 0.5 mM AEBSF, 10 mM sodium butyrate, 5 nM microcystein, 1 mM DTT added fresh) with 0.2% NP-40 on ice for 5 min. The nuclei were collected by centrifuging at 700×g at 4° C. for 5 min. The resulting nuclear pellet was washed twice with the same volume of nuclear isolation buffer without NP-40. Histones were then acid-extracted with 0.2 M H 2 SO 4  for 3 hours at 4° C. with rotation. The insoluble nuclear debris were pelleted at 3400×g at 4° C. for 5 min, and the supernatant was retained. Next, histone proteins were precipitated by adding 100% trichloroacetic acid (TCA) in the ratio of 1:3 (v/v) for 1 hour at 4° C. The pellet was washed with acetone to remove acid residual. Histones were resuspended in 30 μl of 50 mM NH 4 HCO 3  (pH 8.0). 
     Histone Propionylation and Digestion 
     The sample was mixed with 15 μl of derivatization mix, consisting in propionic anhydride and acetonitrile in a ratio of 1:3 (v/v), immediately followed by 7.5 μl of ammonium hydroxide to maintain pH 8.0. The sample was incubated for 15 minutes at 37° C., dried and the derivatization procedure was repeated one more time. Samples were then resuspended in 50 mM NH 4 HCO 3  and incubated with trypsin (enzyme:sample ratio 1:20) overnight at room temperature. After digestion, the derivatization reaction was performed again twice to derivatize peptide N-termini. Samples were desalted using C18 Stage-tips prior to LC-MS analysis. 
     NanoLC-MS/MS 
     Samples were analyzed by using a nanoLC-MS/MS setup. NanoLC was configured with a 75 μm ID×25 cm Reprosil-Pur C18-AQ (3 m; Dr. Maisch GmbH, Germany) nano-column using an EASY-nLC nano-HPLC (Thermo Scientific, San Jose, Calif., USA), packed in-house. The HPLC gradient was as follows: 5% to 32% solvent B (A=0.1% formic acid; B=80% acetonitrile, 0.1% formic acid) over 45 minutes, from 32% to 90% solvent B in 5 minutes, 90% B for 10 minutes at a flow-rate of 300 nL/min. nanoLC was coupled to an Orbitrap Elite mass spectrometer (Thermo Scientific, San Jose, Calif., USA). The acquisition method was data independent acquisition (DIA) as described. Briefly, two full scan MS spectra (m/z 300-1100) were acquired in the ion trap within a DIA duty cycle, and 16 ms/ms were performed with an isolation window of 50 Da. Normalized collision energy (CE) was set to 35%. 
     Data Analysis 
     Raw MS data were analyzed manually. The 7 most intense peptides of histone H3 and H4 containing acetylations were selected, and the relative abundance of the 4th isotope was extracted (compared to the monoisotopic signal). The other peptides were not considered as, due to their low abundance, the relative abundance of all the isotopes could not be confidently quantified. 
     RNA-Sequencing 
     All RNA-seq data were prepared for analysis as follows: NextSeq sequencing data was demultiplexed using native apps on BaseSpace. Demultiplexed FASTQs were aligned by RNA-STAR 2.5.2 to assembly mml0 (GRCm38). Aligned reads were mapped to genomic features using HTSeq 0.6.1. Quantification, library size adjustment, and differential gene expression analysis was done using DESeq2 and Wald&#39;s test (pairwise contrasts between acetate and DMSO-treatment in the inhibitor-treated or untreated cells, followed by a set overlap of differentially expressed genes). Gene list overlaps were tested for significance using the hypergeometric test. 
     Functional Analysis 
     Gene Ontology analysis was performed using DAVID with an FDR cutoff of 10%, filtering categories with fewer than 10 genes or less than 2.5× fold enrichment over background. Motif analysis was performed using HOMER v4.6 on all ACSS2 peaks from published in vivo data (Mews et al., Nature, 2017, 546:381-386) targeting (by the nearest TSS) a gene sensitive to acetate with H3K9ac at the promoter using a fixed window of 300 bp (Mews et al., Nature, 2017, 546:381-386). 
     Mouse Experiments 
     8-week-old adult male mice or E18.5 pregnant females were used. Ethanol, ethanol-d 6 , and sodium acetate-d 3  (Sigma-Aldrich) were administered via intraperitoneal (i.p.) injection and dosed at 2 g/kg (20% solution in saline, filtered through a Stericup GV filter). Conditioned place preference (CPP) was performed according to Cunningham et al. Briefly, Ugo Basile (Italy) mouse CPP boxes (Model Number: 42553) with external dimensions 35×18×29 cm were used. The apparatus was divided into two chambers (16×15×25 cm) that differed in wall and floor pattern. Striped walls were paired with circle cutouts (1 cm) and solid gray walls were paired with square cutouts (0.5 cm). Sessions were run in a dark room at ambient temperature. Boxes were cleaned with 70% ethanol between animals. The paradigm consisted of 1 habituation day (5 min exploration in neutral environment), 1 pre-training session (20 min), 8 training days (biased subject assignment, alternating sessions of saline or ethanol i.p. immediately prior to 5 min session) and 1 post-training test session (20 min). Percent time spent in conditioned chamber was measured. Shapiro-Wilks test was used to assess normal distribution and Mann-Whitney test to determine learning. 
     Intracranial Injection of Viral Vector 
     Adult mice (8+ weeks of age) were anaesthetized with isoflurane gas (1-5% to maintain surgical plane) and placed in a sterile field within a stereotaxic device. Animals received an injection of bupivacaine (2.5 mg kg−1) for local anaesthesia before the skin was disinfected with betadine solution and the skull exposed with a short incision using sterile surgical equipment. Artificial tears were applied to eyes to ensure sufficient lubrication. A small hole (about 0.5 mm) was drilled in the skull over the target area using a stereotax and a stereotactic drill. A micro-syringe filled with viral vector was inserted into the dorsal hippocampus and slowly removed following injection (AP, −2.0 mm; DV, −1.4 mm; 14 ML, ±1.5 mm from bregma). ACSS2 knockdown vector, AAV2/9; U6.shACSS2.CMV.EGFP. All animals received a single dose of subcutaneous meloxicam (5 mg kg) as analgesia at induction and one dose per day for two days postoperatively as needed. 
     The results are now discussed. 
     To determine whether acetate from hepatic alcohol breakdown fuels dynamic histone acetylation in the brain, in vivo stable isotope labeling of protein acetylation was used and monitored by mass spectrometry (MS) (Mews et al., Methods Enzymol., 2016, 574:311-329). Specifically, mice were injected intraperitoneally with 2 g/kg deuterated ethanol (d6-EtOH) or control saline, and deuterium incorporation into acetylated histones was assessed at baseline, as well as 1 and 4 hours after intraperitoneal injections ( FIG. 21A , bottom). Using advanced quantitative liquid chromatography-MS technology, the relative abundance of isotopically labeled histone acetylation in the brain and in peripheral tissues was quantified ( FIG. 21A , top). EtOH-derived acetyl-groups were rapidly incorporated into histone acetylation in the brain, both in hippocampus and in prefrontal cortex ( FIGS. 21B and 21C ,  FIG. 22A-22D ). Label incorporation into histone acetylation was dynamic and heavy labeling decreased to baseline levels 8 hours following intraperitoneal (i.p.) injection. A similar response also occurred in the liver ( FIG. 21D ), which is the principal site of alcohol metabolism, and which has been previously shown to express high levels of ACSS2 (Zakhari, Alcohol Res., 2013, 35:6-16; Comerford et al., Cell, 2014, 159:1591-1602). In contrast, such rapid labeling of histone acetylation was not observed in skeletal muscle (m. gastrocnemius,  FIG. 21E ), which expresses relatively low levels of ACSS2 (Bonthuis et al., Cell Rep., 2015, 12:979-991). 
     To investigate the direct role of ACSS2 in alcohol-dependent acetylation in the brain, ACSS2 expression in the dorsal hippocampus (dHPC) was attenuated by shRNA knockdown using a previously validated viral vector (Mews et al., Nature, 2017, 546:381-386). In these ACSS2 knockdown (KD) animals, heavy-alcohol-derived histone acetylation was compared separately in the dHPC, where ACSS2 was reduced (ACSS2 KD), and in the ventral hippocampus (vHPC), where ACSS2 was normally expressed. Strikingly, ACSS2 KD prevented the incorporation of alcohol-derived heavy acetyl groups into histone acetylation ( FIG. 23A ). In contrast, in the same animal, vHPC incorporation of the heavy label was not affected ( FIG. 23B ). These in vivo data indicate that acetate derived from hepatic alcohol metabolism is transported to the brain and readily incorporated into histone acetylation. Notably, even though the bulk of alcohol metabolism takes place in the liver, alcohol fractions may also be converted to acetate in the brain via the enzymes catalase and CYP2E1 (Zimatkin et al., Alcohol. Clin. Exp. Res., 2006, 30:1500-1505). Therefore, the contribution of extracellular acetate-derived acetyl groups to histone acetylation in the brain was assessed. In mice intraperitoneally injected with 2 g/kg deuterated acetate (d 3 -acetate), rapid label incorporation into brain histone acetylation was detected, at similar levels in both hippocampus and cortex ( FIGS. 23C and 23D ). Remarkably, relative labeling was highest at 30 minutes and returned to background levels at 4 hours post-injection, indicating rapid incorporation of acetate-derived acetyl groups as well as rapid turnover of brain histone acetylation ( FIGS. 23C and 23D ). Together, these data demonstrate that increased blood acetate from liver alcohol metabolism ( FIG. 23E , left) fuels ACSS2-mediated dynamic histone acetylation in the brain ( FIG. 23E , right). 
     Next, the functional relevance of alcohol-derived acetate for ACSS2-dependent histone acetylation in regulating hippocampal gene expression was examined ( FIG. 23E , right). It was found that alcohol administration in wild type (WT) mice resulted in significant enrichment of H3K9ac and H3K27ac peaks genome-wide. Strikingly, this response was eliminated in ACSS2 KD animals significant levels of H3K9ac peaks and a significant amount of H3K27ac peaks failed to induce upon EtOH treatment in the dHPC. RNA-seq was then performed to characterize the transcriptional response and it was found that H3K9ac and H3K27ac drove gene expression in EtOH-treated WT animals genome-wide. However, in line with the ChIP-seq data, this response was markedly blunted in ACSS2 KD mice. Together, the in vivo findings strongly suggest that alcohol administration leads to increased histone acetylation and transcriptional activity in the dHPC in an ACSS2-dependent manner. Because alcohol and acetate have pleiotropic effects on brain circuitry and metabolism, an in vitro assay was then developed to more closely model the direct effects of exogenous acetate on gene expression. Isolated mouse primary hippocampal neurons were utilized to investigate the transcriptional response to supraphysiological levels of acetate (cells were cultured for one week after isolation and subsequently treated with 10 mM acetate for 24 hours) that mimics exogenous acetate influx during alcohol intoxication. Further, to determine the specific role of ACSS2 in transcriptional responses to acetate, a highly specific small molecule inhibitor of ACSS2 (ACSS2i) was employed (Mews et al., Nature, 2017, 546:381-386; Comerford et al., Cell, 2014, 159:1591-1602). 
     In primary hippocampal neurons, acetate supplementation induced 3613 genes ( FIG. 24A ,  FIG. 25A ) that were, via Gene Ontology (GO) term analysis, involved in nervous system processes, including signal transduction and learning and memory ( FIG. 25B , red). In contrast, acetate treatment resulted in down regulation of genes involved in immune system processes ( FIG. 25B , lower in blue). In the presence of the ACSS2i, 2107 of the acetate-induced genes failed to become upregulated ( FIGS. 24A and 24C ), indicating that acetate-induced transcription relies heavily on the catalytic activity of ACSS2. Importantly, acetate-induced genes were not regulated by ACSS2i treatment in the absence of acetate ( FIG. 24B ). GO analysis of ACSS2i-sensitive upregulated genes showed enrichment for nervous system processes, behavior, and learning and memory ( FIG. 24C , lower). Notably, these acetate-induced genes showed a remarkable overlap with genes upregulated by acute alcohol in the hippocampus in vivo (Mulligan et al., Alcohol. Clin. Exp. Res., 2011, 35:659-670) (38% of 214 alcohol-responsive hippocampal genes,  FIG. 25C ), strongly supporting the translational validity of the in vitro model. 
     Further analysis revealed that 40% of the ACSS2i-sensitive upregulated genes are acetylated in vivo (Mews et al., Nature, 2017, 546:381-386) (H3K9ac ChIP-seq, 908 out of 2107 ACSS2i-sensitive genes), and that direct binding of hippocampal ACSS2 (ChIP-seq) is promotor-proximal at baseline without any direct behavioral stimulation in vivo (Mews et al., Nature, 2017, 546:381-386) ( FIG. 23D ). GO analysis linked these genes to intricate plasticity-related mechanisms involving axonogenesis and voltage-gated ion channel activity ( FIG. 24D ). Correspondingly, motif analysis of ACSS2-targeted, acetate-induced, and ACSS2i-sensitive genes implicated the involvement of neuronal transcription factors—including E2F3 and NR5A2 ( FIG. 24E )—that are linked to neurodifferentiation and the regulation of behavior by drugs of abuse (Cates et al., Biol. Psychiatry, 2018, 84:167-179; Stergiopoulos et al., Nat. Commun., 2016, 7:1-16). 
     Together, these findings strongly suggest that ACSS2 plays an important role in alcohol-related learning by coordinating alcohol-induced histone acetylation and gene expression ( FIG. 23C ). To test this behaviorally, ethanol conditioned place preference (CPP) was performed in WT and ACSS2 KD mice. In this paradigm, animals are exposed to neutral and rewarding stimuli in distinct compartments, differentiated by environmental cues. After conditioning, CPP is measured by allowing the animals access to both environments and measuring time spent in the alcohol-associated chamber. In WT mice, rewarding stimuli such as ethanol led to increased time spent in the drug-associated environment (Mann-Whitney, p=0.022). Importantly, the development of CPP depends on dorsal HPC spatial memory formation, and, accordingly, dorsal HPC lesions disrupt place conditioning. Strikingly, it was found that ethanol CPP was completely abolished in ACSS2 KD (dHPC) mice (Mann-Whitney p=0.184), indicating that alcohol-related associative memory formation depends on ACSS2. Taken together, the in vitro, in vivo, and behavioral findings show that ACSS2 is required for heavy labeled acetate incorporation into acetylated histones in the dorsal HPC, which drives memory-related gene expression and alcohol-related associative learning. These results establish ACSS2 as a promising candidate for therapeutic intervention in alcohol use disorders, in which memory of alcohol-associated environmental cues is a primary driver of craving and relapse even after protracted periods of abstinence. 
     Importantly, alcohol exposure not only disrupts epigenetic and transcriptional processes in the adult brain but is also linked to epigenetic dysregulation in the developing fetus (Veazey et al., Epigenetics Chromatin, 2015, 8:39; Starkman et al., Alcohol Research: Current Reviews, 2011, 34:293-305). In utero, alcohol is an environmental teratogen that affects neuro-developmental gene expression and can elicit numerous alcohol-associated postnatal disease phenotypes that are categorized as fetal alcohol spectrum disorder (FASD) (Mead et al., Front. Genet., 2014, 5:1-10). Recent investigation of alcohol-mediated epigenetic changes in utero implicated altered histone acetylation in FASD, but the underlying mechanisms are still unknown (Kim et al., Alcohol Alcohol., 2006, 41:126-132; Mandal et al., Int. J. Biol. Sci., 2017, 13:1100-1108). 
     Whether gestational alcohol exposure may influence histone acetylation in the developing fetal brain was explored via direct deposition of alcohol-derived acetyl groups onto histones. Using the established paradigm of heavy-labeled alcohol injections (2 mg/kg i.p.) followed by mass spectrometry on isolated histone proteins, the incorporation of alcohol metabolites (4 h post-injection) into the neuronal histone acetylation in gestating female mice ( FIG. 26A ) was confirmed, consistent with the previous results in males ( FIGS. 21B and 21C ). Then whether alcohol similarly affects dynamic histone acetylation in utero in the developing mid- and forebrain was investigated (E18.5) ( FIG. 26B ). 
     The fetal brain MS data show that—parallel to maternal labeling of neuronal histone acetylation—‘binge drinking-like’ alcohol exposure resulted in the deposition of alcohol-derived acetyl-groups onto histones in fetal fore- and midbrain at early neural development ( FIG. 26B ). Taken together, the results indicate that direct incorporation of alcohol-derived acetyl groups drives histone acetylation in the fetal brain, indicating an unanticipated potential mechanism for FASD etiology. 
     In the adult brain, epigenetic mechanisms that control gene expression play a key role in processing neural activity. The data provide the first evidence for dynamic signaling from liver alcohol metabolism directly to epigenetic regulation in neurons in vivo, via metabolite activation by neuronal ACSS2. In the hippocampus, alcohol-derived acetyl group incorporation may be of critical importance for alcohol-related associative learning, which encodes environmental cues associated with alcohol that drive craving, seeking, and consumption even after protracted periods of abstinence. Importantly, the findings suggest that other peripheral sources of physiological acetate—primarily the gut microbiome—may similarly affect neuronal histone acetylation and brain function, which may either control or foster other metabolic syndromes. Translational treatment strategies that target this  nexus  between peripheral metabolic activity and neuro-epigenetic regulation may pave the way for novel therapeutic interventions for alcohol use and other neuropsychiatric disorders. 
     Example 4: Small Molecule Inhibition of H3K3 Acetylation 
     Undifferentiated Ntera2 cells were treated with inhibitor for 24 hours with ADG-204, ADG-205 or ADG-206 ( FIG. 27 ). Western blots were used to determine the levels H3K3ac after treatment with ADG-204 ( FIG. 28 ), ADG-205 ( FIG. 29 ) or ADG-206 ( FIG. 30 ). 
     
       
         
         
             
             
         
       
     
     ADG-I-206: 1-(2,3-di(thiophen-2-yl)quinoxalin-6-yl)-3-methylurea (R=Me) 
     To a stirring solution of 2,3-di(thiophen-2-yl)quinoxalin-6-amine (333 mg, 1.08 mmol, 1 eq) in anhydrous CH 2 Cl 2  (5.4 mL) was added N,N-diisopropylethylamine (0.375 mL, 2.15 mmol, 2 eq) followed by triphosgene (105 mg, 0.36 mmol, 0.33 eq) in anhydrous CH 2 Cl 2  (5.4 mL) to give a red-orange solution. The reaction mixture was allowed to stir for 4 h at room temperature, then methylamine (2M in THF, 0.67 mL, 1.35 mmol, 1.25 eq) was added dropwise. The reaction mixture was then allowed to stir for 16 h at room temperature. A stream of argon was blown over the reaction mixture to remove the solvent and any excess phosgene, and the residue obtained was purified by flash chromatography (80% EtOAc/Hexanes) to afford the title compound as a yellow solid (196 mg, 50%).  1 H NMR (500 MHz, DMSO-d 6 ) δ 9.17 (s, 1H), 8.24 (d, J=2.4 Hz, 1H), 7.90 (d, J=9.0 Hz, 1H), 7.84-7.54 (m, 3H), 7.16 (dd, J=13.9, 3.7 Hz, 2H), 7.12-7.06 (m, 2H), 6.29 (q, J=4.6 Hz, 1H), 2.71 (d, J=4.6 Hz, 3H).  13 C NMR (126 MHz, DMSO) δ 155.51, 146.08, 143.15, 142.62, 141.39, 141.19, 141.10, 135.76, 129.65, 129.00, 128.91, 128.72, 128.66, 127.77, 127.65, 123.80, 111.56, 26.29. HRMS (ESI) m/z calc&#39;d for C 18 H 15 N 4 OS 2  [M+H] +  367.0687, found 367.0689 
     ADG-I-205: 1-(2,3-di(thiophen-2-yl)quinoxalin-6-yl)-3-(2-methoxyethyl)urea (R=MeOCH 2 CH 2 ) 
     To a stirring solution of 2,3-di(thiophen-2-yl)quinoxalin-6-amine (337 mg, 1.09 mmol, 1 eq) in anhydrous CH 2 Cl 2  (5.5 mL) was added N,N-diisopropylethylamine (0.38 mL, 2.18 mmol, 2 eq) followed by triphosgene (107 mg, 0.36 mmol, 0.33 eq) in anhydrous CH 2 Cl 2  (5.5 mL) to give a red-orange solution. The reaction mixture was allowed to stir for 4 h at room temperature, then 2-methoxyethylamine (0.12 mL, 1.36 mmol, 1.25 eq) was added dropwise. The reaction mixture was then allowed to stir for 16 h at room temperature. A stream of argon was blown over the reaction mixture to remove the solvent and any excess phosgene, and the residue obtained was purified by flash chromatography (70% EtOAc/Hexanes) to afford the title compound as a yellow solid (196 mg, 50%).  1 H NMR (500 MHz, DMSO-d 6 ) δ 9.19 (s, 1H), 8.23 (d, J=2.4 Hz, 1H), 7.91 (d, J=9.1 Hz, 1H), 7.76 (ddd, J=9.7, 5.1, 1.1 Hz, 2H), 7.68 (dd, J=9.1, 2.4 Hz, 1H), 7.18 (dd, J=3.7, 1.2 Hz, 1H), 7.15 (dd, J=3.7, 1.1 Hz, 1H), 7.10 (ddd, J=6.9, 5.1, 3.7 Hz, 2H), 6.47 (t, J=5.6 Hz, 1H), 3.43 (t, J=5.4 Hz, 2H), 3.33 (t, J=5.5 Hz, 2H), 3.30 (s, 3H).  13 C NMR (126 MHz, DMSO) δ 154.83, 146.12, 143.21, 142.42, 141.37, 141.17, 141.07, 135.79, 129.68, 129.02, 128.94, 128.80, 128.68, 127.77, 127.65, 123.70, 111.55, 71.06, 57.90, 38.87. HRMS (ESI) m/z calc&#39;d for C 20 H 19 N 4 O 2 S 2  [M+H] +  411.0949, found 411.0926. 
     ADG-I-204: 1-(2,3-di(thiophen-2-yl)quinoxalin-6-yl)-3-penlylurea (R=n-C 5 H 1 ) 
     To a stirring solution of 2,3-di(thiophen-2-yl)quinoxalin-6-amine (1.04 g, 3.36 mmol, 1 eq) in anhydrous CH 2 Cl 2  (34 mL) was added N,N-diisopropylethylamine (1.17 mL, 6.72 mmol, 2 eq) followed by triphosgene (329 mg, 1.11 mmol, 0.33 eq) in anhydrous CH 2 Cl 2  (1 mL, final concentration 0.1M) to give a red-orange solution. The reaction mixture was allowed to stir for 4 h at room temperature, then amylmine (0.49 mL, 4.20 mmol, 1.25 eq) was added dropwise. The reaction mixture was then allowed to stir for 16 h at room temperature. A stream of argon was blown over the reaction mixture to remove the solvent and any excess phosgene, and the residue obtained was purified by flash chromatography (50-60% EtOAc/Hexanes) to afford the title compound as a yellow solid (872 mg, 61%).  1 H NMR (500 MHz, DMSO-d 6 ) δ 9.07 (s, 1H), 8.23 (d, J=2.3 Hz, 1H), 7.90 (d, J=9.0 Hz, 1H), 7.76 (dd, J=9.7, 5.0 Hz, 2H), 7.69 (dd, J=9.1, 2.4 Hz, 1H), 7.16 (dd, J=16.6, 3.6 Hz, 2H), 7.13-7.05 (m, 2H), 6.41 (t, J=5.7 Hz, 1H), 3.14 (q, J=6.5 Hz, 2H), 1.48 (p, J=7.1 Hz, 2H), 1.40-1.11 (m, 4H, overlapping with grease), 0.90 (t, J=6.7 Hz, 3H).  13 C NMR (126 MHz, DMSO) δ 154.87, 146.09, 143.13, 142.59, 141.40, 141.19, 141.08, 135.75, 129.67, 129.00, 128.91, 128.74, 128.66, 127.77, 127.65, 123.77, 111.49, 29.26, 28.55, 21.83, 13.91. HRMS (ESI) m/z calc&#39;d for C 22 H23N40S2 [M+H] +  423.1313, found 423.1336. 
     Example 5: Inhibition of Acetyl-CoA Affects Histone Acetylation and Hippocampal Memory 
     To investigate the role of ACSS2 in the adult hippocampus, ACSS2 expression is attenuated in the dorsal hippocampus by treatment with small molecule ACSS2 inhibitors ADG-204, ADG-205, ADG-206 or ADG-207. 
     
       
         
         
             
             
         
       
     
     Compared to control-treated mice, Mice treated with an ACSS2 inhibitor show similar levels of locomotion, coordination, body weight, and anxiety-related thigmotaxis during open field exploration; therefore, ACSS2 inhibition does not cause gross behavioral alterations. 
     To assess hippocampus-dependent spatial memory, an object-location memory paradigm is used. Animals explore three different objects during training, and long-term memory is tested by re-exposure 24 hours later with one object moved to a different location. In training, control and inhibitor treated mice show no difference in exploration. During memory retrieval, control mice show increased exploration of the object that had been moved. By contrast, mice treated with an ACSS2 inhibitor are impaired in spatial object memory and display a lower discrimination index. Mice treated with an ACSS2 show reduced total object exploration during the test, suggesting diminished novelty associated with intact recognition of the objects from the training session. 
     As a control experiment, control mice or mice treated with an ACSS2 inhibitor are subjected to a contextual fear conditioning paradigm. During the 24-hour test session, there are no significant difference in the amount of freezing behavior between control mice or mice treated with an ACSS2 inhibitor suggesting that the ventral hippocampus successfully mediates context-shock association. Overall, ACSS2 has a critical role in dorsal hippocampus-mediated long-term spatial memory. 
     Example 6: Inhibition of Acetyl-CoA Synthetase Prevents the Incorporation of Alcohol-Derived Heavy Acetyl Groups into Histone Acetylation 
     To investigate the direct role of ACSS2 in alcohol-dependent acetylation in the brain, mice are treated with an ACSS2 inhibitor, ADG-204, ADG-205, ADG-206, or ADG-207. Treatment with an ACSS2 inhibitor prevents the incorporation of alcohol-derived heavy acetyl groups into histone acetylation. In contrast, in control mice, vHPC incorporation of the heavy label is not affected. Thus, acetate derived from hepatic alcohol metabolism is transported to the brain and readily incorporated into histone acetylation. 
     Example 7: Synthesis of ADG-207: 1-((1S,3s)-adamantan-1-yl)-3-(2,3-di(thiophen-2-yl)quinoxalin-6-yl)urea (R=1-adamantyl) 
     To a stirring solution of 2,3-di(thiophen-2-yl)quinoxalin-6-amine (32 mg, 0.1 mmol, 1 eq) in anhydrous CH 2 Cl 2  (0.6 mL) was added N,N-diisopropylethylamine (0.04 mL, 0.2 mmol, 2 eq) followed by triphosgene (10 mg, 0.034 mmol, 0.33 eq) in anhydrous CH 2 Cl 2  (0.6 mL, final concentration 0.08 M) to give a red-orange solution. The reaction mixture was allowed to stir for 4 h at room temperature, then 1-adamantanamine (0.49 mL, 4.20 mmol, 1.25 eq) was added dropwise. The reaction mixture was then allowed to stir for 16 h at room temperature. A stream of argon was blown over the reaction mixture to remove the solvent and any excess phosgene, and the residue obtained was purified by flash chromatography (40% EtOAc/Hexanes) to afford the title compound contaminated with 1,1-di-adamantanylurea. The product was re-purified by flash chromatography twice to afford the analytically pure title compound as a yellow solid (4 mg, 8%) 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.91 (s, 1H), 8.20 (d, J=2.4 Hz, 1H), 7.89 (d, J=9.1 Hz, 1H), 7.76 (ddd, J=9.2, 5.1, 1.2 Hz, 2H), 7.61 (dd, J=9.1, 2.4 Hz, 1H), 7.19 (dd, J=3.7, 1.2 Hz, 1H), 7.14 (dd, J=3.7, 1.2 Hz, 1H), 7.10 (ddd, J=11.0, 5.0, 3.6 Hz, 2H), 6.15 (s, 1H), 2.06 (s, 3H), 1.99 (d, J=2.9 Hz, 6H), 1.66 (t, J=3.1 Hz, 6H).  13 C NMR (126 MHz, DMSO) δ 153.55, 146.09, 143.03, 142.57, 141.48, 141.25, 141.06, 135.67, 129.73, 128.98, 128.87, 128.76, 128.65, 127.77, 127.64, 123.66, 111.24, 50.15, 41.50, 36.00, 28.88. HRMS (ESI) m/z calc&#39;d for C 27 H 27 N 4 OS 2  [M+H] +  487.1626, found. 
     The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.