Bromodomain-containing protein 4 (BRD4) is an important epigenetic reader implicated in the pathogenesis of a number of different cancers and other diseases. Brd4-null mouse embryos die shortly after implantation and are compromised in their ability to maintain the inner cell mass, which gives rise to embryonic stem cells (ESCs). Here we report that BRD4 regulates expression of the pluripotency factor Nanog in mouse ESCs and preimplantation embryos, as well as in human ESCs and embryonic cancer stem cells. Inhibition of BRD4 function using a chemical inhibitor, small interfering RNAs, or a dominantnegative approach suppresses Nanog expression, and abolishes the self-renewal ability of ESCs. We also find that BRD4 associates with BRG1 (brahma-related gene 1, aka Smarca4 (SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 4)), a key regulator of ESC self-renewal and pluripotency, in the Nanog regulatory regions to regulate Nanog expression. Our study identifies Nanog as a novel BRD4 target gene, providing new insights for the biological function of BRD4 in stem cells and mouse embryos. Knowledge gained from these non-cancerous systems will facilitate future investigations of how Brd4 dysfunction leads to cancers. Cell Death and Differentiation (2014) 21, 1950-1960 doi:10.1038/cdd.2014; published online 22 August 2014Bromodomain-containing protein 4 (BRD4) belongs to the bromodomain and extraterminal (BET) protein family.1 BRD4 functions as an epigenetic reader by binding to acetylated histones on chromatin through its two bromodomains, and has a central role in transcriptional regulation, cellular growth control and cell cycle progression.2 BRD4 supports transcriptional activation by actively recruiting the positive transcription elongation factor b, mediators and several other transcriptional activators. 2,3 BRD4 is implicated in the pathogenesis of a number of cancers and other diseases. [3][4][5][6][7][8] In some cancers, BRD4 regulates expression of c-Myc and other oncogenes.3,4 It also selectively binds to the 'super-enhancers' of tumor oncogenes, which are large clusters of enhancers that control expression of these genes. 3,9 Although these recent studies have shed light on the gene-specific activity of BRD4, how alterations in BRD4 function contribute to the development of cancers and other diseases is not well understood. This lack of knowledge reflects the need to better understand the normal function of BRD4 in noncancerous cells, as most of the previous studies of BRD4 function were performed in cancer cells.In Brd4 knockout mice, the homozygous embryos die shortly after implantation.10 Cells derived from the inner cell mass (ICM) of these homozygous embryos are completely degenerated, although the rest of the embryo appears morphologically normal. 10 These observations suggest that Brd4 is required for the development and/or maintenance of the ICM, 10 which gives rise to embryonic stem cells (ESCs) in culture. ESCs with homozygous Brd4 deletion are nonviabl...

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Introduction Parkinson’s disease (PD) is a common neurodegenerative disease affecting millions of people world-wide. Fatigue is a prevalent and debilitating non-motor symptom in PD. However, the neural correlates underlying cognitive fatigue are poorly understood. Our previous studies suggested that continuous performance of a simple but mentally demanding psychomotor vigilance task (PVT) induced cognitive fatigue, operationalized as subjective exhaustion and time-on-task performance decline. Here we used arterial spin labeling (ASL) perfusion fMRI to investigate regional cerebral blood flow (CBF) changes in PD patients during cognitive fatigue induced by continuous performance of 20-min PVT. Methods Twenty-one PD patients completed a 20-min PVT during the ASL scan and two additional 4-min resting-state ASL scans before and after PVT. Reaction times (RTs) and regional CBF changes throughout the PVT as well as during pre- and post-task resting baselines were measured. Cognitive fatigue was analyzed by dividing the entire PVT performance into five quintiles in addition to the immediate measurement of self-rated fatigue before and after PVT. Results PD patients demonstrated significantly increased self-reported fatigue ratings after the task (p < 0.05) and progressively slower RTs across quintiles (p < 0.05). Perfusion data showed that the PVT activates the right middle frontal cortex, right inferior parietal lobe, right insula, bilateral occipital cortex, and right cerebellum (FDR corrected). Moreover, the bilateral middle frontal gyri were less active during the post-task rest compared to the pre-task rest. Conclusion These results demonstrated that cognitive fatigue has an ongoing effect on brain activity after a period of continuous mental effort and supported the critical role of prefrontal cortex in mediating cognitive fatigue in PD. The findings also suggest the utility of continuous PVT as an appropriate paradigm to induce and examine cognitive fatigue in PD. Support Supported in part by Parkinson’s Foundation Translational Research Grant and NIH grants R01-MH107571, R21-AG051981, and P30-NS045839.

Introduction The locus coeruleus (LC) plays a key role in the regulation of arousal and autonomic function. Homeostatic sleep pressure refers to the drive for sleep that increases as a saturating exponential when we stay awake and decreases exponentially when we sleep. The current study used arterial spin labeling (ASL) functional magnetic resonance imaging (fMRI) to investigate the relationship between homeostatic sleep pressure (sleepiness) and LC activity during the psychomotor vigilance test (PVT). Methods We analyzed sleepiness and ASL imaging data from N=70 health adults (40 males, age range 21–50 years) who participated in a controlled in-laboratory sleep study. All participants were scanned at rest and during the PVT on the morning between 0700h-1000h after 9 hour time-in-bed (TIB) baseline sleep. LC regions-of-interest (ROI) were defined by standard templates from Keren et al. (2009). Sleepiness was assessed by the Karolinska Sleepiness Scale (KSS) every two hours from 10:30 am to 10:30 pm. Results Sleepiness scores gradually increased over wakefulness time and reached its peak in the evening at about 10:20pm. PVT-induced CBF changes did not correlate with sleepiness scores on the morning (p > 0.05), but showed significant negative correlations with sleepiness scores on later day when sleep pressure became higher, especially during the night-time (r = -0.41, p < 0.001). Specifically, LC CBF showed significant increases during the PVT scan as compared to the resting scan (p = 0.04) in individuals with less nigh-time sleepiness (KSS < 4), but no differences (p > 0.1) in individuals with greater nigh-time sleepiness (KSS ≥ 5). After controlling for age, gender, and total sleep time, PVT-induced regional CBF difference in the LC still negatively predicted sleepiness (β = -0.325, p = 0.005). Conclusion Our findings showed that individuals with greater LC CBF increases during the PVT were less sleepy during the night, supporting the key role of LC activity in promoting wakefulness and maintaining sleep homeostasis. PVT-induced LC activation may provide a non-invasive bio-marker of homeostatic sleep pressure in healthy adults. Support Supported in part by NIH grants R01-HL102119, R01-MH107571, R21-AG051981. CTRC UL1RR024134, and P30-NS045839.

Introduction Locus coeruleus (LC) is the major source of norepinephrine (NE) in the brain, which plays a key role in maintaining arousal and alertness. Sleep loss significantly impairs arousal and alertness. However, it is unknown whether sleep loss disrupts LC integrity, which can be measured non-invasively by diffusion tensor imaging (DTI). In the current study, we used DTI to examine the effects of one night of acute total sleep deprivation (TSD) on fractional anisotropy (FA), an index reflecting fiber density, axonal diameter and myelination. Methods We analyzed DTI and psychomotor vigilance test (PVT) data from N=54 health adults (23 females, age range 21–50 years) from a well controlled in-laboratory sleep deprivation study. Participants were randomized to either a TSD condition (n=40) without sleep on night 2, or a control condition (n=14) with no sleep loss. Standard DTI scans were conducted on the morning of day 2 and day 3 between 0700h-1000h. The PVT reciprocal response time (RRT) was used to measure individual’s response speed at baseline without sleep loss. LC regions-of-interest (ROI) were defined by standard templates from Keren et al. (2009). Imaging data were analyzed using FSL toolbox. Results For the whole TSD group, no differences were found in the LC FA values before and after sleep deprivation (p > .2). However, when dividing the TSD group to a slow group and a fast group based on their baseline PVT response speed, significantly increased LC FA were found in the slow group (p = .007) but not in the fast group (p > .4). The PVT RRT negatively correlated with LC FA value changes after TSD (r = -.44, p = .004). No correlations were found between the PVT RRT and LC FA changes in the control group. Conclusion Our results showed that baseline vigilance response speed correlated with LC integrity change after sleep deprivation, with slower response exhibiting greater changes in LC integrity. These findings support the key role of LC-NE system in the regulation of alertness and arousal. Support Supported in part by NIH grants R01-HL102119, R01-MH107571, R21-AG051981. CTRC UL1RR024134, and P30-NS045839.