Clinical trial NCT06690866: Negative Emotionality and Epigenetics During Puberty
Official title: Negative Emotionality in Relation to Epigenetics of Estrogen Signaling During Puberty
Study type: OBSERVATIONAL
Overall status: RECRUITING
Conditions: Puberty
Healthy
Stress
Keywords: Stress reactivity
DNA methylation
fMRI
Lead sponsor: International Research Training Group 2804
Sponsor class: OTHER
Collaborators: name: University Hospital Tuebingen
name: Uppsala University
name: German Research Foundation
First posted: 2024-11-15
Enrollment: 100
Sex: FEMALE
Minimum age (years): 8
Maximum age (years): 17
Standard ages: CHILD
Accepts healthy volunteers: yes
Observational model: COHORT
Time perspective: CROSS_SECTIONAL
Sampling method: NON_PROBABILITY_SAMPLE
Study population: Pubescent girls between 8 and 17 years of age living and schooling in Tübingen and surrounding area

Brief summary: Pubertal transition leads to enduring neuroendocrine changes along with changes in the epigenome. The prevalence of psychiatric disorders significantly increases in females compared to males after puberty. There is likely to be an interaction between epigenetics, hormones and neurophysiological processes during puberty, leading to the increased prevalence of mental disorders in females. This study aims to shed light on these interactions underlying the emerging sex differences after puberty. Specifically, it seeks to investigate the epigenetic modifications and subsequent changes in gene expression during the pubertal transition and their association with negative emotionality (e.g., acute stress response and depressive symptoms) at molecular, neuronal, subjective and physiological levels.

Detailed description: In order to investigate epigenetic and neuroimaging correlates of negative emotionality in pubertal girls, the study employs a cross-sectional design, enrolling 50 pre-pubescent girls (8-10 years) and 50 post-pubescent girls (15-17 years) as healthy participants. Comprehensive data will be collected using self- and parent-report questionnaires on pubertal status, prenatal complications, adverse life events, stress and coping mechanisms, mood and anxiety symptoms, gender identity, and reproductive health. To delve into the neuronal correlates of stress, participants will undergo (f)MRI and MIST. Additionally, physiological stress responses will be assessed through heart rate and skin conductance measurements. Participants will provide blood, saliva, and hair samples for hormone, methylation and expression analysis. DNA methylation analysis will be performed on whole blood and saliva samples using pyrosequencing, focusing on genes that are responsive to estrogen and genes involved in estrogen signaling, based on the previous literature reporting an overrepresentation of estrogen-responsive genes among the differentially methylated sites across the entire genome in girls during puberty. The effects of methylation on gene expression will be measured using reverse transcription real-time PCR. Cortisol levels will be assessed from saliva collected six times during MIST to evaluate the acute stress response and from hair to assess chronic stress. Sex steroids such as estrogen and progesterone from blood will be analyzed using LC-MS/MS. The study aims to provide insights into the intricate relationship between epigenetic modifications, gene expression and negative emotionality (e.g., stress reactivity and internalizing symptoms) during puberty in girls.

Eligibility criteria: Inclusion Criteria:

* healthy girls
* aged between 8-10 and having pubertal stage 1 or between 15-17 and having pubertal stage 5
* normal body mass index according to age (between 5th and 85th percentile)
* non-smoking
* German language fluency
* Attending age-appropriate schools

Exclusion Criteria:

* neurological or psychiatric disease
* medical problems such as hormonal, metabolic, developmental or chronic diseases (e.g., congenital disorders, precocious puberty, polycystic ovarian syndrome, diabetes or congestive heart failure)
* any kind of hormonal, pharmacological or psychotropic treatment in the last three months
* engaging in competitive/extreme sports

Additional exclusion criteria for MRI:

* People with non-removable metal objects on or in the body
* Tattoos (if not MRI-incompatible according to expert guidelines)
* Pathological hearing or increased sensitivity to loud noises
* Claustrophobia
* Surgery less than three months ago
* Moderate or severe head injury

Primary outcomes:
measure: DNA methylation differences between pre- and post-pubertal girls in candidate genes
timeFrame: Measured once after MRI measurement (approximately 30 minutes).
measure: Correlation between DNA methylation and neuronal activity during acute stress
timeFrame: Measured once: 3 runs of MIST lasting 20 minutes in total.
measure: The mediating role of sex steroids in DNA methlation and negative emotionality
timeFrame: Measured once for each part: 20 minutes for MIST, 30 minutes for blood and saliva collection, and 1 hour for questionnaires
Secondary outcomes:
measure: Correlation between DNA methylation and gene expression in candidate genes
timeFrame: Measured once after MRI measurement (approximately 30 minutes)
measure: Correlation between DNA methylation and HPA-axis response
timeFrame: Measured six times: 1 hour before MIST(1), just before(1) and after(1) MIST, and three times more in 20-minute intervals after MIST. Each saliva collection lasts approx. 2 minutes.
measure: Correlation between DNA methylation and subjective stress response
timeFrame: Measured six times: 1 hour before MIST(1), just before(1) and after(1) MIST, and three times more in 20-minute intervals after MIST. Each rating takes approx. 10 minutes.
measure: Correlation between DNA methylation and phsiological stress response
timeFrame: Measured once: 20 minutes for phsiological data during MRI measurement
measure: Correlation between DNA methylation and functional connectivity
timeFrame: Measured once with resting-state functional MRI, approximately 10 mintes
measure: Correlation between DNA methylation and brain gray matter structure
timeFrame: Measured once with MPRAGE MRI, approximately 10 minutes. Analyzed and reported over the course of the study, in average of 1 year
Other outcomes:
Locations:
city: Tuebingen
country: Germany
facility: University of Tuebingen; Department of Psychiatry & Psychotherapy; Tuebingen
state: BW
Study officials:
affiliation: University Hospital Tuebingen
name: Vanessa Nieratschker, Prof.
role: PRINCIPAL_INVESTIGATOR
affiliation: University Hospital Tuebingen
name: Mirac Nur Musaoglu, MD
role: STUDY_DIRECTOR
References: