DNA methylation of the oxytocin receptor gene predicts neural response to ambiguous social stimuli

“The following pre-statistics processing was then applied: motion correction using MCFLIRT (Jenkinson et al., 2002); non-brain removal using BET (Smith, 2002); spatial smoothing using a Gaussian kernel of FWHM 5 mm; grand-mean intensity normalization of the entire 4D dataset by a single multiplicative factor; highpass…”

“The following pre-statistics processing was then applied: motion correction using MCFLIRT (Jenkinson et al., 2002); non-brain removal using BET (Smith, 2002); spatial smoothing using a Gaussian kernel of FWHM 5 mm; grand-mean intensity normalization of the entire 4D dataset by a single multiplicative factor; highpass…”

“The following pre-statistics processing was then applied: motion correction using MCFLIRT (Jenkinson et al., 2002); non-brain removal using BET (Smith, 2002); spatial smoothing using a Gaussian kernel of FWHM 5 mm; grand-mean intensity normalization of the entire 4D dataset by a single multiplicative factor; highpass…”

“Despite direct anatomical connections between dACC and the limbic system, most traditional neurocognitive models posit a role for dACC in executive functions such as modulation of attention and cognitive control (Bush et al., 2000).”

“, 2009)], and more recent evidence indicates its involvement in activation of transcription (Lister et al., 2009; as reviewed in Suzuki and Bird, 2008).”

“…widely discussed role in sexual behavior (Carter, 1992), maternal behavior (Feldman et al., 2007), and affiliative functions like promoting trust (Kosfeld et al., 2005), pair bonding (Ross and Young, 2009), and encoding of positive social stimuli (Guastella et al., 2008a), OXT also appears to be…”

“To assess this prediction, we utilized a classic social perception task first developed by Heider and Simmel (1944) and later adapted by Castelli et al. (2000) in which simple geometric shapes either move and interact in ways that imply animacy [the Animate condition (ANIM)] or display an identical…”

“However, more recent evidence suggests that this region plays a critical role in social and affective appraisals of motivationally salient stimuli (Etkin et al., 2011).”

“…NEUROSCIENCE in differentiation and development, is commonly associated with transcriptional silencing [in some cases heritable (Kaminsky et al., 2009)], and more recent evidence indicates its involvement in activation of transcription (Lister et al., 2009; as reviewed in Suzuki and Bird, 2008).”

“At first level, time-series statistical analysis was carried out using fMRIB’s software library improved linear model (FILM) with local autocorrelation correction (Woolrich et al., 2001).”

“This brain region is adjacent to the temporal parietal junction and near the anterior portion of the superior temporal sulcus (STS), which has been strongly implicated in social cognition by virtue of its role in biological motion perception (Allison et al., 2000; Pelphrey and Morris, 2006) and mentalizing (Frith and…”

“, 2000; Pelphrey and Morris, 2006) and mentalizing (Frith and Frith, 2003; Gallagher and Frith, 2003) functions.”

“, 2000; Pelphrey and Morris, 2006) and mentalizing (Frith and Frith, 2003; Gallagher and Frith, 2003) functions.”

“Higher-level analysis of the data was conducted with a mixed-effects approach using fMRIB’s local analysis of mixed effects (FLAME) stage 1 (Beckmann et al., 2003; Woolrich et al., 2004; Mark, 2008), which allows for the carrying up of first-level fixed effects variances to the higher-level analysis and forces the RAN…”

“…OXT administration increases time spent looking at the eyes when viewing human faces (Guastella et al., 2008b), improves recognition of emotions conveyed by the eyes (Domes et al., 2007), and increases sensitivity to biological motion embedded in a noisy background (Kéri and Benedek, 2009).”

“This task reliably recruits a network of brain structures believed to be important for social perception and mentalizing abilities, including the temporal parietal junction (Castelli et al., 2000; Ross and Olson, 2010).”

“In general, although participants are given no further instructions than to watch the stimuli, this task appears to elicit not just perception of animacy, but also attribution of intentionality and social-emotional content to the interactions between the geometric shapes (Castelli et al., 2000; Scholl and Tremoulet, 2…”

“, 2005), pair bonding (Ross and Young, 2009), and encoding of positive social stimuli (Guastella et al.”

“A whole-brain analysis was conducted; Z (Gaussianised T/F) statistic images were thresholded using clusters determined by Z > 2.3 and a (corrected) cluster significance threshold of p < 0.05 (Worsley, 2001).”

“…resources such as optimism and self-esteem (Saphire-Bernstein et al., 2011), non-verbal intelligence (Lucht et al., 2009), behavioral and dispositional empathy (Rodrigues et al., 2009), positive affect (Lucht et al., 2009), and parental sensitivity (BakermansKranenburg and van IJzendoorn, 2008).”

“ORIGINAL RESEARCH ARTICLE published: 10 October 2012 doi: 10.3389/fnhum.2012.00280 DNA methylation of the oxytocin receptor gene predicts neural response to ambiguous social stimuli”

“Twin studies have indicated that methylation level at specific CpG sites are more commonly shared by twins when compared to unrelated individuals, are correlated between tissues and can be stable across the life span (Kaminsky et al., 2009; Bell et al., 2012).”

“While traditional models of epigenetic inheritance proposed erasure of DNA methylation in non-imprinted regions between generations, contemporary reports in model systems and humans suggest that in some cases this epigenetic modification may survive reprogramming (Daxinger and Whitelaw, 2012; Labrie et al., 2012).”

“in differentiation and development, is commonly associated with transcriptional silencing [in some cases heritable (Kaminsky et al., 2009)], and more recent evidence indicates its involvement in activation of transcription (Lister et al.”

“In humans, the actions of OXT have been reported to be mediated by a receptor that is encoded by the oxytocin receptor gene (OXTR) (Kimura et al., 1992).”

“This SNP is also associated with structural differences in oxytonergic brain regions including amygdala and hypothalamus, as well as functional differences during emotional face processing (Tost et al., 2010).”

“…significantly greater activity in the ANIM versus RAND condition, were generally consistent with previous literature (Castelli et al., 2000; Schultz et al., 2003): the posterior STS bilaterally, the right precentral gyrus, the left inferior frontal gyrus, the dorsomedial prefrontal cortex,…”

“Activity in pSTS, for example, has previously been found to increase in situations in which more elaborated processing of moving social stimuli is required, such as when a character’s actions are incongruent with expectations (Pelphrey et al., 2003, 2004).”

“Such limitations may be addressed, at least in part, by influential new models of human disease that suggest that expression of risk-related genes could be controlled by epigenetic processes (Bjornsson et al., 2004).”

“Moreover, abnormal function of dACC has been associated with emotional and social perceptual deficits in anxiety disorders (McClure et al., 2007) and ASD (Dichter et al., 2009).”

“The first variant, rs2254298 (G>A), is associated with autism spectrum disorders (ASD) (Jacob et al., 2007) and unipolar depression (Costa et al., 2009).”

“In addition to its more widely discussed role in sexual behavior (Carter, 1992), maternal behavior (Feldman et al., 2007), and affiliative functions like promoting trust (Kosfeld et al., 2005), pair bonding (Ross and Young, 2009), and encoding of positive social stimuli (Guastella et al., 2008a),…”

“There have been documented differences in global levels of DNA methylation as a function of ethnicity and gender (Zhang et al., 2011).”

“ORIGINAL RESEARCH ARTICLE published: 10 October 2012 doi: 10.3389/fnhum.2012.00280 DNA methylation of the oxytocin receptor gene predicts neural response to ambiguous social stimuli”

“Activity in pSTS, for example, has previously been found to increase in situations in which more elaborated processing of moving social stimuli is required, such as when a character’s actions are incongruent with expectations (Pelphrey et al., 2003, 2004).”

“ORIGINAL RESEARCH ARTICLE published: 10 October 2012 doi: 10.3389/fnhum.2012.00280 DNA methylation of the oxytocin receptor gene predicts neural response to ambiguous social stimuli”

“The second variant, rs53676 (G > A), is associated with decreases in psychological resources such as optimism and self-esteem (Saphire-Bernstein et al., 2011), non-verbal intelligence (Lucht et al.”

“This task reliably recruits a network of brain structures believed to be important for social perception and mentalizing abilities, including the temporal parietal junction (Castelli et al., 2000; Ross and Olson, 2010).”

“While tissue-specific variation in DNA methylation has been observed, recent work indicates that, on the whole, methylation patterns are relatively conserved across tissue types within individuals (Byun et al., 2009).”

“Additionally, the degree to which areas that are involved in mentalizing processes are activated by these displays is dependent upon the degree to which participants attend to the intentional, contingent relationship relative to mechanical trajectories of the shapes (Blakemore et al., 2003).”

“If higher degrees of DNA methylation lead to decreased transcription of OXTR in the brain (Kusui et al., 2001; Gregory et al., 2009) one might expect processes mediated by OXT to be more perturbed in individuals high in DNA methylation.”

“Carriers of the risk allele of OXTR SNP rs2254298 have increased dACC volume relative to non-risk allele carriers (Furman et al., 2011).”

“The first variant, rs2254298 (G>A), is associated with autism spectrum disorders (ASD) (Jacob et al., 2007) and unipolar depression (Costa et al., 2009).”

“Moreover, in some cases, epigenetic modifications assessed peripherally have been demonstrated to serve as useful predictors of the same modifications in the phenotypically relevant tissue such as the brain (Kaminsky et al., 2011).”

“While traditional models of epigenetic inheritance proposed erasure of DNA methylation in non-imprinted regions between generations, contemporary reports in model systems and humans suggest that in some cases this epigenetic modification may survive reprogramming (Daxinger and Whitelaw, 2012; Labrie et al., 2012).”

“…OXT administration increases time spent looking at the eyes when viewing human faces (Guastella et al., 2008b), improves recognition of emotions conveyed by the eyes (Domes et al., 2007), and increases sensitivity to biological motion embedded in a noisy background (Kéri and Benedek, 2009).”

“Similarly, given the role of dACC in resolving ambiguity in expressed emotion (Nomura et al., 2003), a main effect of methylation in this region could indicate that persons with higher levels of OXTR methylation perceive the presented ANIM interactions as more ambiguous, and consequently recruit dACC more heavily in a…”

“Moreover, activity in the adjacent left STS region is sensitive to attributions of causality of biological motion (Morris et al., 2008).”

“…from computerized games and other forms of behavioral paradigms have implicated oxytocin in trust (Kosfeld et al. 2005), empathy (Carter et al. 2009b), cooperation (Hurlemann et al. 2010, Rilling et al. 2012) and neural activation in brain regions associated with sociality ( Jack et al. 2012).”

“In neurotypical humans, methylation status of the OXTR also has been shown to strongly predict measures of social perception in both college-aged adults and infants (Jack et al., 2012; Puglia et al., 2015, 2018; Krol et al., 2019b).”

“Altered plasma levels of OXT and AVP have been reported in autistic individuals and they are often correlated to aberrant functional connectivity [110,111].”

“OXTR mRNA expression is affected by methylation of promoter, and high levels of methylation have been associated with ASD.(252,256) Consistent with this, a study on siblings and adults with ASD found increased OXTR promoter methylation.”

“Linking DNA methylation with a particular ASD-associated endophenotype, Jack and colleagues found a strong correlation between OXTR promoter methylation in PBMCs and perception of animacy (purposeful movement) of an inanimate object, with correlated neural responses in two brain regions measured by functional magnetic…”

“…the review by Kumsta et al. (in press) shows that changes to methylation levels in the promoter region CpG island are associated with reductions in mRNA signaling of the gene (Kusui et al., 2001) and related to phenotypes of autism (Gregory et al., 2009) and social cognition (Jack et al., 2012).”

“DNA methylation within the promoter region of OXTR is variable within the population (36, 37), and methylation of specific OXTR CpG sites reduces transcription of the gene (38).”

“, anterior cingulate cortex) (123,127) or predominantly left hemisphere (122,125,126,128,130) and right cerebellum effects (Supplement) (127).”

“The first reported that DNA methylation at the OXTR was predictive of an individual’s neural response to biological motion (Jack et al., 2012).”

“We extracted probes located within the OXTR CpG island (n = 12), as this area has been previously investigated by Dadds et al (10; see Figure 1, Panel A) and others (13, 21, 22), and shown to play a key role in modulating the transcriptional activity of OXTR (23).”

“…whole blood has been associated with increased fMRI BOLD signal in the left superior temporal gyrus/supramarginal gyrus and right dorsal anterior cingulate cortex during a task that involved perception of ‘animacy’ and the ability to attribute mental states to geometric shapes (Jack et al, 2012).”

“These include positive associations between OXTR DNAm and callous-unemotional traits in youth [47, 58], social cognitive deficits in ASD [45, 46], rigid thinking in anorexia nervosa [49], affect regulation problems [41, 57] and mood deficits [38–40] as well as limbic regions linked with facial and emotional recognitio…”

“Though definitive data that relate blood and brain methylation at CpG site -934 are not available, we have previously shown the utility of this assay in prediction of brain endophenotypes of social perception (Jack et al., 2012; Puglia et al., 2015).”

“More recently, human experiments support the hypothesis that dynamic changes in components of the OT system (i.e., methylation of the OTR gene; Jack et al., 2012; Unternaehrer et al., 2012) and possibly neurodevelopmental changes in OT sensitive brain structures (see Andari et al.”

“This effect would alter the individual’s vulnerability to the effects of stress (Jack et al., 2012).”

“For OXTr rs53576, it has been demonstrated that A carriers, who are considered more vulnerable compared to G carriers, display reduced hypothalamic volume, that is shown to be related to susceptibility in ASD development (Liu et al., 2010; Tost et al., 2010; Jack et al., 2012; Wang et al., 2013).”

“OXTR hypermethylation was associated with enhanced neural activity to negative emotions in frontal areas of infants [39] and with BOLD activity in temporal areas and ACC when attributing intentionality and socioemotional content to geometric shapes [23].”

“The level of DNA methylation of the OXTR gene is associated with variations in social perception in patients with autistic spectrum disorders [22].”

“However, based on effect sizes reported in studies investigating other oxytocin genes (19, 20, 46, 47), namely OXTR, and behavior or brain associations, we estimated that a sample size of 66 or larger would be adequate to test each of our hypotheses.”

“These findings also lead to an important question: are the social processes under epigenetic control? [26].”

“DNA methylation in MT2 is associated with neural endophenotypes in typical populations: DNA methylation is positively correlated with brain activity during animacy perception [27], emotion perception [28, 29], and social attention [30].”

“The OT receptor gene OXTR has been reported to be associated with ASD and an association between the degree of OXTR methylation and brain activity evoked by the perception of animacy has been reported (Jack et al. 2012; Jacob et al. 2007).”

“of social perception and individual variability in social perception (displays of animacy) (Jack et al., 2012; Puglia et al., 2015).”

“…family with autism (Gregory et al., 2009), and DNA methylation of a single CpG in this promoter region in peripheral blood mononuclear cells (PBMCs) has since been associated with altered neural activity in multiple brain regions in functional MRI scans (Jack et al., 2012; Puglia et al., 2015).”

“Pyrosequencing was performed as previously described [55].”

“First, we provide novel evidence that by five months of age, human infants vary extensively in the epigenetic modification of OXTR , similar to the variability found in adults (Gregory et al., 2009; Puglia et al., 2015; Jack et al., 2012).”

“In line with the previous literature (Bell al., 2014; Dadds et al., 2014; Gregory et al., 2009; Jack et al., 2012; Puglia et al., 2015), our data suggests that low OXTR methylation and high plasma OT levels are associated with better social outcomes, (i.”

“Increased methylation at residue –934 in the promoter of the OXTR gene correlated with an increased BOLD fMRI signal in the temporal parietal junction of human subjects performing behavioral tasks that are related to social perception (Jack et al. 2012).”

“, 2014) and increased brain activity during a social task (Jack et al., 2012).”

“Although we do not know the exact association between blood and brain methylation at CpG site -934, a number of studies to date reflect the utility of this peripheral assay in predicting brain endophenotypes related to social cognition (34, 35).”

“Methylation status in the OXTR CpG island was shown to be functional in healthy humans: Higher levels of methylation were associated with higher neural activation in STG, SMG and dorsal ACC in a social perception task [Jack et al., 2012].”

“In humans, DNA methylation of the OXTR in blood cells predicted the neural response to ambiguous social stimuli and rapidly changed following acute psycho-social stress (Jack et al., 2012; Unternaehrer et al., 2012).”

“Another study showed hypermethylation of the OXTR promoter in fetal membrane from preterm birth, a strongly associated condition with ASD incidence, implying that OXTR methylation in fetus could be a surrogate marker of ASD (Behnia et al., 2015).”

“Epigenetic variation in methylation of the OXTR gene is similarly associated with neural responses to ambiguous social stimuli (Jack et al., 2012), autism and callousunemotional traits (Gregory et al.”

“Finally epigenetic studies show that different methylation patterns in OXTR are involved in the predisposition to classic autism [31] and variability of social perception [32].”

“Thus, OTR methylation differences at the CpG site (-934) in peripheral blood cells have been correlated with different levels of brain activity in the temporal parietal junction and in the dorsal anterior cingulate cortex ( Jack et al. 2012).”

“However, this traditional view has been recently challenged, with more and more studies finding an inverse relationship, highlighting the complexities in predicting behavioral phenotypes from DNA methylation (Jack et al., 2012; Reiner et al., 2015; Ziegler et al., 2015).”

“ASD individuals have a higher level of OXTR methylation and, consequently, a reduced expression of the OXTR gene.(19) Some of the other candidate genes in ASD are the Methyl CpG binding Protein 2 gene, BCL-2, Retinoic Acid-Related Orphan Receptor Alpha (RORA), SHANK3, maternally expressed gene UBE3A, and engrailed-2 (…”

“High methylation of the OXTR promoter has been linked to particular neurological patterns (Jack et al., 2012).”

“To evaluate PCR amplification and interrogation of BSC saliva DNA, we used a well-established assay for the oxytocin receptor (OXTR) (chr3: 8,792,095 to 8,811,300: GRCh37/hg19 build), which has used in studies of psychiatric disorders [52–54] and social behaviors [55,56].”

“Other studies that examined OXTR genotype within typical populations usually implicate areas of the social brain other than the rSMG, such as the amygdala or the ACC [50, 67–69, 90].”

“, 2004) and their receptors (Francis et al., 2000; Kusui et al., 2001; Champagne et al., 2001; Bester-Meredith and Marler, 2001; Kimura et al., 2003; Gregory et al., 2009; Jack et al., 2012).”

“Moreover, DNA methylation of OTR assessed in blood cells correlates with higher BOLD activity in the anterior cingulate cortex and superior temporal gyrus, two brain regions important for social perception (Jack et al., 2012).”

“For the OXTR, methylation differences have been associated with functional changes in the brain (Jack et al., 2012), and are a likely candidate for the differences in brain structure and function that are related to differences in these SNPs.”

“…pSTS, temporo-parietal junction (TPJ), and medial prefrontal cortex (mPFC) (Castelli et al. 2000; Martin and Weisberg 2003; Schultz et al. 2003; Ohnishi et al. 2004; Gobbini et al. 2007; Pavlova et al. 2010; Ross and Olson 2010; Tavares et al. 2011; Jack et al. 2012; Osaka, Ikeda, and Osaka 2012).”

“As a consequence, epigenetic dysregulations in these brain neuropeptide systems could play a role in social dysfunctions in ASDs, suggesting that social processes might be in some measure under epigenetic control [64,65].”

“…data from blood samples, indicates that DNA methylation of the oxytocin receptor gene might predicts neural response to ambiguous social stimuli (Jack et al., 2012) and also, it was recently demonstrated that epigenetic modification of the oxytocin receptor gene could influence social cognition…”

“Recent neuroimaging evidence indicates, for the first time, that degree of methylation of the oxytocin receptor gene predicts neural response within the left superior temporal gyrus and the anterior cingulate gyrus in response to animate social movement compared with random social movement.(59) Hence, not only does so…”

“Specifically, higher OXTR methylation at site −934 is associated with increased activity in brain regions implicated in social processing, and with decreased functional coupling of the amygdala within regions supporting social perception and emotion processing [61,62].”

“Extremely high levels of methylation may predict lower activity in the areas associated with social perception (Jack et al., 2012).”

“For example, methylation of the human MT2 region has been associated with autism,(19) high callousunemotional trait,(20) stress reactivity(21) and altered neural responses to face stimuli.(22,23) Therefore, peripheral MT2 methylation may provide information about OXTR expression in the brain that has functional signif…”

“Three CpG sites in the 5′ UTR of OXTR exon 2 (OXTR1) and eight CpG sites in the protein coding region of OXTR exon 3 (OXTR2) were targeted based on previous studies: OXTR1 (Chr3: 8,810,729–8,810,845; GRCh37/hg19) [30] and OXTR2 (Chr3: 8,809,281–8,809,534; GRCh37/hg19) [11, 40].”

“, 2012 [86] PBMCs MT2 -934 Bisulfitepyrosequencing BOLD response to ambiguous social stimuli DNA methylation was positively associated with BOLD response to ambiguous social stimuli in left superior temporal gyrus and cingulate gyrus.”

“In particular, OXTR methylation has been linked to autism (Gregory et al., 2009), social perception (Jack et al., 2012), callous-unemotional traits (Dadds et al., 2014), anxiety/depression (Chagnon et al., 2015) as well as anger and fear perception (Puglia et al., 2015).”

“Interestingly, DNA methylation signatures were also associated with brain volume (41, 42), brain structure and function (22, 43, 44), as well as social (45), and cognitive functioning (46), even though the studies were performed in peripheral tissues (blood/saliva).”

“The methylation of the oxytocin receptor gene was reported to increase the activity in the temporal parietal junction, which is known to participate in social perception [71].”

“Moreover, a brain imaging study reported higher Oxtr methylation associated with increased brain activity in cortical areas involved in social perception [118].”

“In humans methylation status of the OXTR also has been shown to predict neural responses to ambiguous social stimuli (Jack et al. 2012).”

“However, in humans, OTalso acts as a neuromodulator in neural networks associated with trust (Kosfeld et al. 2005; for replication problems, see Nave et al. 2015), empathy (Decety and Batson 2009), and sociality (Jack et al. 2012).”