Selected Publications

Is oxytocin receptor signaling really dispensable for social attachment?

Joshua S Danoff, Emma A Whelan, Jessica J Connelly. Comprehensive Psychoneuroendocrinology. May 2023.

Our commentary on Berendzen et al.'s paper examining oxytocin receptor mutants in prairie voles.

An epigenetic mechanism for differential maturation of amygdala-prefrontal connectivity in choldhood socio-emotional development

Amalia M Skyberg, Benjamin T Newman, Andrew J Graves, Alison M Goldstein, Samantha R Brindley, Minah Kim, Jason Druzgal, Jessica J Connelly, James P Morris. Translational Psychiatry. March 13, 2023

Functional connectivity between the amygdala and the medial prefrontal cortex (mPFC) has been identified as a neural substrate of emotion regulation that undergoes changes throughout development, with a mature profile typically emerging at 10 years of age. Maternal bonding in childhood has been shown to buffer amygdala reactivity and to influence the trajectory of amygdala–mPFC coupling. The oxytocinergic system is critical in the development of social behavior and maternal bonding. Early-life parental care influences the methylation status of the oxytocin receptor (OXTRm) in animal models and humans, and higher OXTRm is associated with lower amygdala–PFC functional connectivity in adults. Using a neuroimaging-epigenetic approach, we investigated saliva-derived OXTRm as a biological marker of structural and functional connectivity maturation in 57 typically developing children (P < 0.05). We utilized seed-based connectivity analysis during a novel abstract movie paradigm and find that higher levels of OXTRm are associated with a more adult-like functional connectivity profile. Concurrently, more adult-like functional connectivity was associated with higher reported self-control and more diffusion streamlines between the amygdala and mPFC. OXTRm mediates the association between structural and functional connectivity with higher levels of OXTRm being associated with more streamlines. Lastly, we also find that lower OXTRm blunts the association between amygdala–mPFC connectivity and future internalizing behaviors in early adolescence. These findings implicate OXTRm as a biological marker at the interface of the social environment and amygdala–mPFC connectivity in emotional and behavioral regulation. Ultimately, identification of neurobiological markers may lead to earlier detection of children at risk for socio-emotional dysfunction.

Adolescent peer struggles predict accelerated epigenetic aging in midlife

Joseph P Allen, Joshua S Danoff, Meghan A Costello, Emily L Loeb, Alida A Davis, Gabrielle L Hunt, Simon G Gregory, Stephanie N Giamberardino, Jessica J Connelly. Development and Psychopathology. 2022 April 5.

This study examined struggles to establish autonomy and relatedness with peers in adolescence and early adulthood as predictors of advanced epigenetic aging assessed at age 30. Participants (N = 154; 67 male and 87 female) were observed repeatedly, along with close friends and romantic partners, from ages 13 through 29. Observed difficulty establishing close friendships characterized by mutual autonomy and relatedness from ages 13 to 18, an interview-assessed attachment state of mind lacking autonomy and valuing of attachment at 24, and self-reported difficulties in social integration across adolescence and adulthood were all linked to greater epigenetic age at 30, after accounting for chronological age, gender, race, and income. Analyses assessing the unique and combined effects of these factors, along with lifetime history of cigarette smoking, indicated that each of these factors, except for adult social integration, contributed uniquely to explaining epigenetic age acceleration. Results are interpreted as evidence that the adolescent preoccupation with peer relationships may be highly functional given the relevance of such relationships to long-term physical outcomes.

Lifetime marijuana use and epigenetic age acceleration: A 17-year prospective examination

Joseph P Allen, Joshua S Danoff, Meghan A Costello, Gabrielle L Hunt, Amanda F Hellwig, Kathleen M Krol, Simon G Gregory, Stephanie N Giamberardino, Karen Sugden, Jessica J Connelly. Drugs and Alcohol Dependence. 2022 April 1.

Aims

This study was designed to assess links between lifetime levels of marijuana use and accelerated epigenetic aging.

Design

Prospective longitudinal study, following participants annually from age 13 to age 30.

Setting and participants

A community sample of 154 participants recruited from a small city in the Southeastern United States.

Measurements

Participants completed annual assessments of marijuana use from age 13 to age 29 and provided blood samples that yielded two indices of epigenetic aging (DNAmGrimAge and DunedinPoAm) at age 30. Additional covariates examined included history of cigarette smoking, anxiety and depressive symptoms, childhood illness, gender, adolescent-era family income, and racial/ethnic minority status.

Findings

Lifetime marijuana use predicted accelerated epigenetic aging, with effects remaining even after covarying cell counts, demographic factors and chronological age (β’s = 0.32 & 0.27, p’s < 0.001, 95% CI’s = 0.21–0.43 & 0.16–0.39 for DNAmGrimAge and DunedinPoAm, respectively). Predictions remained after accounting for cigarette smoking (β’s = 0.25 & 0.21, respectively, p’s < 0.001, 95% CI’s = 0.14–0.37 & 0.09–0.32 for DNAmGrimAge and DunedinPoAm, respectively). A dose-response effect was observed and there was also evidence that effects were dependent upon recency of use. Effects of marijuana use appeared to be fully mediated by hypomethylation of a site linked to effects of hydrocarbon inhalation (cg05575921).

Conclusions

Marijuana use predicted epigenetic changes linked to accelerated aging, with evidence suggesting that effects may be primarily due to hydrocarbon inhalation among marijuana smokers. Further research is warranted to explore mechanisms underlying this linkage.

Neuroepigenetic impact on mentalizing in childhood

Amalia M Skyberg, Stefen Beeler-Duden, Alison M Goldstein, Christina A Gancayco, Angeline S Lillard, Jessica J Connelly, James P Morris. Developmental Cognitive Neuroscience. 2022 Geb 8.

Mentalizing, or the ability to understand the mental states and intentions of others, is an essential social cognitive function that children learn and continue to cultivate into adolescence. While most typically developing children acquire sufficient mentalizing skills, individual differences in mentalizing persist throughout childhood and are likely influenced by a combination of cognitive functioning, the social environment, and biological factors. DNA methylation of the oxytocin receptor gene (OXTRm) impacts gene expression and is associated with increased brain activity in mentalizing regions during displays of animacy in healthy young adults. The establishment, fine-tuning, and implications of such associations in the context of broader social functioning remain unclear. Using a developmental neuroimaging epigenetic approach, we investigated the contributions of OXTRm to individual variability in brain function during animate motion perception in middle childhood. We find that higher levels of OXTRm are associated with increased neural responses in the left temporo-parietal junction and inferior frontal gyrus. We also find a positive association between neural activity in LTPJ and social skills. These findings provide evidence of epigenetic influence on the developing child brain and demonstrate that variability in neural social perception in childhood is multifaceted with contributions from individual social experience and the endogenous oxytocin system.

An epigenetic rheostat of experience: DNA methylation of OXTR as a mechanism of early life allostasis

Joshua S Danoff, Jessica J Connelly, James P Morris, Allison M Perkeybile. Comprehensive Psychoneuroendocrinology. 2021 Nov 14.

Oxytocin is a neuropeptide hormone which is involved in regulation of social behavior, stress response, muscle contraction, and metabolism. Oxytocin signaling is dependent on its binding to the oxytocin receptor, coded for by the OXTR gene. Many studies have examined the role of epigenetic regulation of OXTR in neurological and behavioral outcomes in both humans and animal models. Here, we review these studies, critically analyze their findings in the context of oxytocin’s role as an allostatic hormone, and provide suggestions for future research. We use OXTR as a model for how those in the field of psychoneuroendocrinology should perform epigenetic studies in order to maximize both biological relevance and potential for biomarker development.

Genetic, epigenetic, and environmental factors controlling oxytocin receptor gene expression

Joshua S Danoff, Kelly L Wroblewski, Andrew J Graves, Graham C Quinn, Allison M Perkeybile, William M Kenkel, Travis S Lillard, Hardik I Parikh, Hudson F Golino, Simon G Gregory, C. Sue Carter, Karen L Bales, Jessica J Connelly. Clinical Epigenetics. 2021 Jan 30.

Background
The neuropeptide oxytocin regulates mammalian social behavior. Disruptions in oxytocin signaling are a feature of many psychopathologies. One commonly studied biomarker for oxytocin involvement in psychiatric diseases is DNA methylation at the oxytocin receptor gene (OXTR). Such studies focus on DNA methylation in two regions of OXTR, exon 3 and a region termed MT2 which overlaps exon 1 and intron 1. However, the relative contribution of exon 3 and MT2 in regulating OXTR gene expression in the brain is currently unknown.
Results
Here, we use the prairie vole as a translational animal model to investigate genetic, epigenetic, and environmental factors affecting Oxtr gene expression in a region of the brain that has been shown to drive Oxtr related behavior in the vole, the nucleus accumbens. We show that the genetic structure of Oxtr in prairie voles resembles human OXTR. We then studied the effects of early life experience on DNA methylation in two regions of a CpG island surrounding the Oxtr promoter: MT2 and exon 3. We show that early nurture in the form of parental care results in DNA hypomethylation of Oxtr in both MT2 and exon 3, but only DNA methylation in MT2 is associated with Oxtr gene expression. Network analyses indicate that CpG sites in the 3′ portion of MT2 are most highly associated with Oxtr gene expression. We also identify two novel SNPs in exon 3 of Oxtr in prairie voles and a novel alternative transcript originating from the third intron of the gene. Expression of the novel alternative transcript is associated with genotype at SNP KLW2.
Conclusions
These results identify putative regulatory features of Oxtr in prairie voles which inform future studies examining OXTR in human social behaviors and disorders. These studies indicate that in prairie voles, DNA methylation in MT2, particularly in the 3′ portion, is more predictive of Oxtr gene expression than DNA methylation in exon 3. Similarly, in human temporal cortex, we find that DNA methylation in the 3′ portion of MT2 is associated with OXTR expression. Together, these results suggest that among the CpG sites studied, DNA methylation of MT2 may be the most reliable indicator of OXTR gene expression. We also identify novel features of prairie vole Oxtr, including SNPs and an alternative transcript, which further develop the prairie vole as a translational model for studies of OXTR.

Epigenetic tuning of brain signal entropy in emergent human social behavior

Meghan H Puglia, Kathleen M Krol, Manuela Missana, Cabell L Williams, Travis S Lillard, James P Morris, Jessica J Connelly, Tobias Grossmann. BMC Medicine. 2020 Aug 17.

Background: How the brain develops accurate models of the external world and generates appropriate behavioral responses is a vital question of widespread multidisciplinary interest. It is increasingly understood that brain signal variability-posited to enhance perception, facilitate flexible cognitive representations, and improve behavioral outcomes-plays an important role in neural and cognitive development. The ability to perceive, interpret, and respond to complex and dynamic social information is particularly critical for the development of adaptive learning and behavior. Social perception relies on oxytocin-regulated neural networks that emerge early in development.
Methods: We tested the hypothesis that individual differences in the endogenous oxytocinergic system early in life may influence social behavioral outcomes by regulating variability in brain signaling during social perception. In study 1, 55 infants provided a saliva sample at 5 months of age for analysis of individual differences in the oxytocinergic system and underwent electroencephalography (EEG) while listening to human vocalizations at 8 months of age for the assessment of brain signal variability. Infant behavior was assessed via parental report. In study 2, 60 infants provided a saliva sample and underwent EEG while viewing faces and objects and listening to human speech and water sounds at 4 months of age. Infant behavior was assessed via parental report and eye tracking.
Results: We show in two independent infant samples that increased brain signal entropy during social perception is in part explained by an epigenetic modification to the oxytocin receptor gene (OXTR) and accounts for significant individual differences in social behavior in the first year of life. These results are measure-, context-, and modality-specific: entropy, not standard deviation, links OXTR methylation and infant behavior; entropy evoked during social perception specifically explains social behavior only; and only entropy evoked during social auditory perception predicts infant vocalization behavior.
Conclusions: Demonstrating these associations in infancy is critical for elucidating the neurobiological mechanisms accounting for individual differences in cognition and behavior relevant to neurodevelopmental disorders. Our results suggest that an epigenetic modification to the oxytocin receptor gene and brain signal entropy are useful indicators of social development and may hold potential diagnostic, therapeutic, and prognostic value.

OXTR DNA methylation moderates the developmental calibration of neural reward sensitivity

Marlen Z Gonzalez, Kelly L Wroblewski, Joseph P Allen, James A Coan, Jessica J Connelly. Developmental Psychobiology. 2020 August 17.

The Adaptive Calibration Model of Stress Responsivity (ACM) suggests that developmental experiences predictably tune biological systems to meet the demands of the environment. Particularly important is the calibration of reward systems. Using a longitudinal sample (N = 184) followed since adolescence, this study models the dimensions of early life stress and their effects on epigenetic modification of the oxytocin receptor gene (OXTR) and individual differences in neural response to reward anticipation. We first created a latent variable model of developmental context using measures collected when participants were 13 years old. As adults, two subsets of participants completed a reward anticipation fMRI paradigm (N = 82) and agreed to have their blood assayed for (OXTR) DNA methylation (N = 112) at two CpG sites. Three latent constructs of developmental context emerged: Neighborhood Harshness, Family Harshness, and Abuse and Disorder. Greater OXTR DNA methylation at CpG sites -924 and -934 blunted the association between greater Neighborhood Harshness and increased neural activation in caudate in anticipation of rewards. Interaction effects were also found outside of reward-related areas for all three latent constructs. Results indicate an epigenetically derived differential susceptibility model whereby high methylation coincides with decreased association between developmental environment and neural reward anticipation.

Epigenetic dynamics in infancy and the impact of maternal engagement

Krol KM, Moulder RG, Lillard TS, Grossmann T, Connelly JJ. Science Advances. 2019 October 16.

The contribution of nature versus nurture to the development of human behavior has been debated for centuries. Here, we offer a piece to this complex puzzle by identifying the human endogenous oxytocin system-known for its critical role in mammalian sociality-as a system sensitive to its early environment and subject to epigenetic change. Recent animal work suggests that early parental care is associated with changes in DNA methylation of conserved regulatory sites within the oxytocin receptor gene (OXTRm). Through dyadic modeling of behavior and OXTRm status across the first year and a half of life, we translated these findings to 101 human mother-infant dyads. We show that OXTRm is dynamic in infancy and its change is predicted by maternal engagement and reflective of behavioral temperament. We provide evidence for an early window of environmental epigenetic regulation of the oxytocin system, facilitating the emergence of individual differences in human behavior.

Behavioral and epigenetic consequences of oxytocin treatment at birth

Kenkel WM, Perkeybile AM, Yee JR, Pournajafi-Nazarloo H, Lillard TS, Ferguson EF, Wroblewski KL, Ferris CF, Carter CS, Connelly JJ. Science Advances. 2019 May 1.

Oxytocin is used in approximately half of all births in the United States during labor induction and/or augmentation. However, the effects of maternal oxytocin administration on offspring development have not been fully characterized. Here, we used the socially monogamous prairie vole to examine the hypothesis that oxytocin exposure at birth can have long-term developmental consequences. Maternally administered oxytocin increased methylation of the oxytocin receptor (Oxtr) in the fetal brain. As adults, oxytocin-exposed voles were more gregarious, with increased alloparental caregiving toward pups and increased close social contact with other adults. Cross-fostering indicated that these effects were the result of direct action on the offspring, rather than indirect effects via postnatal changes in maternal behavior. Male oxytocin-exposed offspring had increased oxytocin receptor density and expression in the brain as adults. These results show that long-term effects of perinatal oxytocin may be mediated by an epigenetic mechanism.

Epigenetic modification of the oxytocin receptor gene is associated with emotion processing in infant brain.

Krol KM, Puglia MH, Morris JP, Connelly JJ, Grossmann T. Developmental Cognitive Neuroscience. 2019 June.

The neural capacity to discriminate between emotions emerges early in development, though little is known about specific factors that contribute to variability in this vital skill during infancy. In adults, DNA methylation of the oxytocin receptor gene (OXTRm) is an epigenetic modification that is variable, predictive of gene expression, and has been linked to autism spectrum disorder and the neural response to social cues. It is unknown whether OXTRm is variable in infants, and whether it is predictive of early social function. Implementing a developmental-neuroimaging-epigenetics approach in a large sample of infants (N=98), we examined whether OXTRm is associated withneural responses to emotional expressions. OXTRm was assessed at 5 months of age. At 7 months of age, infants viewed happy, angry, and fearful faces while functional near-infrared spectroscopy was recorded. We observed that OXTRm shows considerable variability among infants. Critically, infants with higher OXTRm show enhanced responses to anger and fear and attenuated responses to happiness in right inferior frontal cortex, a region implicated in emotion processing through action-perception coupling. Findings support models emphasizing oxytocin’s role in modulating neural response to emotion and identify OXTRm as an epigenetic mark contributing to early brain function.

Early nurture epigenetically tunes the oxytocin receptor.

Perkeybile AM, Carter CS, Wroblewski KL, Puglia MH, Kenkel WM, Lillard TS, Karaoli T, Gregory SG, Mohammadi N, Epstein L, Bales KL, Connelly JJ. Psychoneuroendocrinology. 2018 Aug 31;99:128-136

Mammalian sociality is regulated in part by the neuropeptide oxytocin. In prairie voles, subtle variation in early life experience changes oxytocin receptor-mediated social behaviors. We report that low levels of early care in voles leads to de novo DNA methylation at specific regulatory sites in the oxytocin receptor gene (Oxtr), impacting gene expression and protein distribution in the nucleus accumbens. DNA methylation state of the blood predicts expression in the brain indicating the utility of the blood as a biomarker for the transcription state of the brain. These experience-sensitive CpG sites are conserved in humans, are related to gene expression in the brain, and have been associated with psychiatric disorders and individual differences in neural response to social stimuli. These results identify a mechanism by which early care regulates later displays of typical prairie vole social behavior and suggest the potential for nurture driven epigenetic tuning of OXTR in humans.

Epigenetic regulation of the oxytocin receptor is associated with neural response during selective social attention

Puglia MH, Connelly JJ, Morris JP. Translational Psychiatry. 2018 Jun.

Aberrant attentional biases to social stimuli have been implicated in a number of disorders including autism and social anxiety disorder. Oxytocin, a naturally-occurring mammalian hormone and neuromodulator involved in regulating social behavior, has been proposed to impact basic biological systems that facilitate the detection of and orientation to social information. Here, we investigate a role for naturally-occurring variability in the endogenous oxytocinergic system in regulating neural response during attention to social information. Participants performed a selective social attention task while undergoing fMRI, provided a blood sample for epigenetic analysis, and completed self-report measures of social functioning. We find that a functional epigenetic modification to the oxytocin receptor, OXTR methylation, is associated with increased neural response within and decreased functional coupling between regions of the salience and attentional control networks during selective social attention. We also show that subclinical variability in autistic and social anxiety traits moderates this epigenetic regulation of neural response. These data offer a mechanistic explanation to a growing literature associating social behavior and disorder with epigenetic modification to OXTR by suggesting that OXTR methylation reflects a decrease in the extent to which social information automatically captures attention. We highlight the importance that treatment efficacy be considered in relation to individual differences in molecular makeup, and that future studies aimed at uncovering biomarkers of disorder carefully consider measurement at both the biological and phenotypic level.