Biological Response to Brief Psychological Challenge

Transduction of Psychological Stress Into Systematic Inflammation by Mitochondrial DNA Signaling

The investigators plan to conduct a crossover experimental trial examining physiological responses to a socio-evaluative speech task under laboratory conditions. Participants will attend two laboratory sessions. At one session participants will take part in a brief laboratory stress task and at the other participants will rest for the same period. Measures of cardiovascular response will be assessed at both sessions. In addition, blood will be drawn at multiple time points across a 125 minute period to assess changes in circulating levels of cortisol, catecholamines, markers of inflammation and cell free mitochondrial DNA in response to the task. The investigators expect that the stress task will induce a specific increase in ccf-mtDNA, which will statistically mediate subsequent peak circulating Interleukin-6 and Tumor Necrosis Factor-α levels. In secondary analyses, the investigators will examine whether stress-induced increases in circulating cortisol, epinephrine, and norepinephrine levels correlate with increases in ccf-mtDNA. These studies will establish the kinetics and magnitude of psychological stress-induced ccf-mtDNA release, the association with early stress mediators, and whether ccf-mtDNA mediates the inflammatory response to acute stress in humans.

Study Overview

• Status: Completed
• Conditions: Acute Inflammatory Response to Psychological Stress
• Intervention / Treatment: Behavioral: Socio-evaluative speech task; Behavioral: Control, Quiet Rest

Detailed Description

The proposed study will examine physiologic responses to acute psychological challenge in the laboratory among healthy adults. It is widely accepted that there is an increase in circulating markers of inflammation following a single bout of laboratory stress. This increase in systemic inflammation is believed to contribute to the damaging health effect of psychological stress. However, to date, the biological mechanisms by which psychological stress is transduced into inflammation are unclear. The investigators' preliminary evidence suggests that mitochondrion may play a role, with stress-induced increases in circulating levels of mitochondria- derived signaling molecules that are known to modulate immune cell function and the production of pro-inflammatory cytokines.

To test this possibility, the investigators plan to conduct a crossover experimental trial examining physiological responses to an evaluative speech task under laboratory conditions. The investigators have previously used this task to induce physiological arousal. The investigators plan to recruit 60 non-smoking volunteers (50% female, aged 20-50 years) and test these participants on two occasions separated by at least a month. On one occasion the participants will be exposed to the speech task. On the other occasion, the participants will rest quietly for the same period. Conditions will be counterbalanced. At both visits cardiovascular responses (heart rate, blood pressure, and heart rate variability) will be assessed as measures of autonomic activation before, during and after the task period. Participants will also have an intravenous catheter inserted and blood drawn at ten time points over the two hour testing period on each occasion. Blood samples will be sent to laboratories at the University of Pittsburgh and at Columbia University for the assessment of mitochondria-derived signalling molecules, inflammatory markers, and cortisol levels.

• Study Type: Interventional
• Enrollment (Actual): 72
• Phase: Not Applicable

Contacts and Locations

Study Locations

• United States
  • Pennsylvania
    • Pittsburgh, Pennsylvania, United States, 15213
      • University of Pittsburgh

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 16 years to 46 years (Adult)
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Generally healthy
• Non-smokers/illicit drug users
• Blood pressure below 140/90
• Weight > 110 lbs
• BMI < 30
• Fluent in English
• Women -- regular menstrual cycles over the past 12 months (defined as 21- 35 days in length)
• Able and willing to give informed consent
• Willing to abstain from alcohol and vigorous exercise for 24 hours, from food and drinks (other than water) for 3 hours and from non-prescription medications (other than oral contraception) for 2 days before testing.
• Willing to attend two laboratory stress testing sessions, give blood though an intravenous catheter, undergo medical evaluation and complete psychosocial questionnaires.

Exclusion Criteria:

• Reported history of chronic systemic immune, metabolic or mitochondrial diseases, or chronic diseases that influence the central nervous, autonomic nervous or neuroendocrine systems, e.g., autoimmune disease, chronic infections, cardiovascular disease, diabetes, chronic kidney or liver disease, cancer treatment.
• Reported psychiatric history of schizophrenia or other psychotic illness, or mood disorder.
• Resting blood pressure > 140/90 mmHg at baseline testing.
• Weight < 110 lbs
• BMI equal to or greater than 30
• Report currently taking glucocorticoid, anti-inflammatory, anti-retroviral, immunosuppressant, insulin, antiarrhythmic, antihypertensive, oral hypoglycemic, antidepressant, benzodiazepine or prescription weight loss medications or other medications known to influence the immune, autonomic or neuroendocrine systems.
• For women - Post-menopausal or irregular menstrual cycles over the past 12 months. Report current pregnancy or lactation.
• Current smokers (defined as having smoked a cigarette in the previous 3 months).
• Current illicit drug use (defined as reported use of illicit drugs such as marijuana, cocaine or heroin in the previous 3 months).
• Not fluent in English (have used English in everyday speaking and reading for at least 10 years)
• Unable or unwilling to give informed consent
• Unwilling to abstain from alcohol and vigorous exercise for 24 hours, from food and drinks (other than water) for 3 hours and from non-prescription medications (other than oral contraception) for 2 days prior to testing.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Single

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Socio-evaluative Speech Stress, then Control; Participants will attend two laboratory sessions. At the first session, participants will complete a socio-evaluative speech task, which is a widely used, highly effective way to investigate stress responses in a laboratory setting. Participants will prepare and deliver a brief, 3-minute speech defending themselves against an alleged transgression (e.g., running a stop sign). The speech will be delivered in front of a video camera, a mirror and an audience (the interviewer and another staff member). Participants will be told that their non-verbal behaviors are being evaluated. At the second session, participants will rest quietly for the same period as the speech task, in the absence of the stressor.	Behavioral: Socio-evaluative speech task; 5-minute speech task designed to induce physiological arousal in a laboratory setting.; Behavioral: Control, Quiet Rest; 5-minute quiet rest period.
Experimental: Control, then Socio-Evaluative Speech Stress; Participants will attend two laboratory sessions. At the first session, participants will rest quietly for 5 minutes. At the second session, participants will complete a socio-evaluative speech task, which is a widely used, highly effective way to investigate stress responses in a laboratory setting. Participants will prepare and deliver a brief, 3-minute speech defending themselves against an alleged transgression (e.g., running a stop sign). The speech will be delivered in front of a video camera, a mirror and an audience (the interviewer and another staff member). Participants will be told that their non-verbal behaviors are being evaluated.	Behavioral: Socio-evaluative speech task; 5-minute speech task designed to induce physiological arousal in a laboratory setting.; Behavioral: Control, Quiet Rest; 5-minute quiet rest period.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Cell-free Mitochondrial DNA	Serum levels of mitochondrial DNA assessed from blood samples	5 minutes before task, and 5, 10, 20, 30, 45, 60, 75, 90 and 120 minutes after the task.
Interleukin-6	Plasma levels of interleukin-6	5 minutes before to 5, 10, 20, 30, 45, 60, 75, 90, 120 post-task periods
Tumor Necrosis Factor-alpha	Plasma levels of tumor necrosis factor-alpha	5 minutes before to 5, 10, 20, 30, 45, 60, 75, 90, 120 post-task periods

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Heart Rate	Continuous measurement of heart rate was averaged across 4 periods: last 5 minutes of baseline, 5-min task-period, first two 5 minutes post-task.	last 5 minutes of baseline, 5-min task-period, first two 5 minutes post-task.
Systolic Blood Pressure	Blood pressure was assessed twice on 10 occasions across the protocol. The two readings on each occasion were averaged.	5 min pre-task and 5, 10, 20, 30, 45, 60, 75, 90, 120
Diastolic Blood Pressure	Diastolic blood pressure was assessed two times on 10 occasions across the protocol. On each occasion, the two measures were averaged.	5 min pre-task and 5, 10, 20, 30, 45, 60, 75, 90, 120 post task onset
Cortisol	Circulating levels of cortisol assessed by ELISA	5 minutes before to 10, 20, 30, 45, 60 minutes post-task periods
Epinephrine	Levels of epinephrine in plasma	5 minutes before to 5, 10, 20, 30, & 60 minutes post-task periods
Norepinephrine	Levels of norepinephrine in plasma	5 minutes before to 5, 10, 20, 30, & 60 minutes post-task periods
Heart Rate Variability	Interbeat intervals of heart rate assessed by 3-lead EKG. Measures were taken continuously from 5 minutes before the task to 10 minutes after the task. Rsults were then averaged across 4 periods: 5 minutes prior to the task, the 5-minute task period, and 5-, and 10-minutes post task	Pre-task, task, and 1-5 and 6-10 minutes post task
Fatigue	Momentary assessment of fatigue, measured as score on the fatigue subscale on the brief Profile of Mood States questionnaire. Scores range from 0 - 20, with higher scores reflecting more fatigue.	2 minutes before and 2, 60, and 120 minutes post-task periods
Anger	Momentary assessment of anger, measured as score on the anger subscale on the brief Profile of Mood States questionnaire in response to the task periods. Scores range from 0 - 12, with higher scores reflecting more anger.	2 minutes before and 2-, 60-, and 120-minutes post-task periods
Anxious Mood	Momentary assessment of anxious mood, measured as score on the anxiety subscale on the brief Profile of Mood States questionnaire in response to the task periods. Scores range from 0 - 16, with higher scores reflecting more anxious mood.	2 minutes before and 2-, 60- and 120-minutes post-task periods
Depressed Mood	Momentary assessment of depressed mood, measured as score on the depression subscale on the brief Profile of Mood States questionnaire in response to the task periods. Scores range from 0 - 12, with higher scores reflecting more depressed mood.	2 minutes before and 2-, 60- and 120-minutes post-task periods
Vigor	Momentary assessment of vigor, measured as score on the vigor subscale on the brief Profile of Mood States questionnaire in response to the task periods. Scores range from 0 - 12, with higher scores reflecting more vigor.	2 minutes before and 2-, 60-, and 120-minutes post-task periods
Wellbeing	Momentary assessment of wellbeing, measured as score on the wellbeing subscale on the brief Profile of Mood States questionnaire in response to the task periods. Scores range from 0 - 12, with higher scores reflecting more wellbeing.	2 minutes before and 2-, 60- and 120-minutes post-task periods
Calm Mood	Momentary assessment of calm mood, measured as score on the calm subscale on the brief Profile of Mood States questionnaire in response to the task periods. Scores range from 0 - 16, with higher scores reflecting more calm mood.	2 minutes before and 2-, 60-, and 120-minutes post-task periods

Collaborators and Investigators

• Sponsor: University of Pittsburgh
• Collaborators: National Institute of Mental Health (NIMH)
• Investigators: Principal Investigator: Anna L Marsland, Ph.D., University of Pittsburgh

Publications and helpful links

General Publications

• Trumpff C, Marsland AL, Basualto-Alarcon C, Martin JL, Carroll JE, Sturm G, Vincent AE, Mosharov EV, Gu Z, Kaufman BA, Picard M. Acute psychological stress increases serum circulating cell-free mitochondrial DNA. Psychoneuroendocrinology. 2019 Aug;106:268-276. doi: 10.1016/j.psyneuen.2019.03.026. Epub 2019 Mar 28.
• Trumpff C, Marsland AL, Sloan RP, Kaufman BA, Picard M. Predictors of ccf-mtDNA reactivity to acute psychological stress identified using machine learning classifiers: A proof-of-concept. Psychoneuroendocrinology. 2019 Sep;107:82-92. doi: 10.1016/j.psyneuen.2019.05.001. Epub 2019 May 7.

Study record dates

Study Major Dates

• Study Start (Actual): July 23, 2020
• Primary Completion (Actual): January 31, 2022
• Study Completion (Actual): January 31, 2022

Study Registration Dates

• First Submitted: August 28, 2019
• First Submitted That Met QC Criteria: September 3, 2019
• First Posted (Actual): September 4, 2019

Study Record Updates

• Last Update Posted (Actual): March 1, 2024
• Last Update Submitted That Met QC Criteria: February 27, 2024
• Last Verified: February 1, 2024

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Behavioral Symptoms; Stress, Psychological
• Other Study ID Numbers: STUDY19020140; R01MH119336 (U.S. NIH Grant/Contract)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: Individual deidentified participant data (including data dictionaries) will be shared. This includes all of the individual -participant data collected during the trial. In addition, the study protocol will be made available. The Co-Principal Investigators of the study will oversee all matters related to data management and archiving. Final research data will be deposited in the digital repository of the Inter-university Consortium for Political and Social Research (ICPSR), of which the University of Pittsburgh is a member, and which receives partial funding from the NIH.
• IPD Sharing Time Frame: Data will be made available within nine months to one year after the publication of results bearing on the project's specific aims. The Inter-university Consortium for Political and Social Research (ICPSR) will archive the full dataset and its documentation for the long term, supporting the data through changing technologies, new media, and data formats.
• IPD Sharing Access Criteria: All publications and presentations deriving from the final research data will include information on the location of the data and how it can be accessed, well as acknowledgement of the repository and funding source. The ICPSR has policies and procedures in place that will provide data access to qualified researchers, fully consistent with NIH data sharing policies and applicable laws and regulations.
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No