Multi-level Molecular Profiling of High Acute Stress: a Clinical Study

Study Overview
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Brief Summary
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Although it is well known that stress plays an important role in the development of neuropsychiatric diseases, the precise role and molecular effects of stress have only been poorly understood. For example, autophagy is essential for energy and cellular homeostasis through protein catabolism, and dysregulation results in compromised proteostasis, stress-coping behavior, and excessive secretion of signaling molecules and inflammatory factors. Therefore, the aim of the project is to analyze the clinical effects of a bungee jump resembling an acute stress event in correlation to autophagy and other underlying, multi-level molecular profiling. Specifically, it is planned to perform multi-level molecular profiling and sleep analysis in a cohort of healthy male individuals before, during, and after a bungee jump compared to a control cohort of healthy males not undergoing a stress event. The resulting findings will advance the role of autophagy during the stress response and hence in the development of psychiatric disorders, and possibly investigate alternative treatment venues on a molecular level, and finally contribute to a better clinical outcome.

Detailed Description
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Although it is well known that stress plays an important role in the development of neuropsychiatric diseases, the molecular effects of stress have only been poorly understood. So far it is known that stress leads to an activation of the stress hormone axis followed by an increased release of the stress hormone and glucocorticoid cortisol. Glucocorticoids bind to glucocorticoid receptors that initiate a cellular signal cascade. However, it can be assumed that other factors are involved but a profound understanding of the stress response at the molecular level has not yet been performed yet. Using a so-called multi-omics approach it is possible to determine changes in a large number of molecular groups, such as proteins or lipids to research the underlying mechanisms of diseases. While multi-omics analyzes have already helped gain elementary knowledge in a large number of somatic diseases, the molecular effects of acute stress have not been addressed yet. This will be the primary focus of this study. To achieve this an acute, concise stress reaction closely resembling a genuine stress response is desired. In previous studies, it was shown that bungee jumping triggers such a short, intense stress reaction and the corresponding activation of the stress hormone axis. To achieve this a cohort of 25-30 healthy male individuals who undergo a bungee jump resembling an acute stress event will be compared to a cohort of 10-20 healthy males who undergo the same experimental design without undertaking a bungee jump or other stress intervention. At different time points (baseline, shortly before and after the intervention, at multiple time points during the intervention as well as around one week follow up after the intervention) serval psychometrical questionnaires will be gathered and blood will be collected. A dexamethasone inhibition test will be performed before the stress intervention. Sleep quality will be additionally assessed during the entire course of the study by actigraphy. On selected days blood will be collected. Following, autophagy activity will be assessed by Western Blot analysis, and mass spectrometry-based proteomics, phosphoproteomics, metabolomics, and lipidomics will be performed. Bioinformatic analysis, statistical evaluation, quality control, and in silico pathway analyses will then specifically identify factors and cascades of relevance. The aim of the project is to analyze the clinical effects of an acute stress event in correlation to the underlying, multi-level molecular profiling. Longitudinal multi-omic profiling including proteome, metabolome, lipidome, and epigenetic changes will reveal time-series analysis of thousands of molecular changes and an orchestrated composition of autophagy depended signaling. The resulting findings will advance the role of autophagy in the development of psychiatric disorders, and possibly investigate alternative treatment venues on a molecular level, and finally contribute to a better clinical outcome.

Official Title
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Multi-level Molecular Profiling of High Acute Stress

Conditions
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Healthy

Intervention / Treatment
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* Behavioral: Bungee jump


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Present written declaration of consent Healthy Male BMI between 18,5 and 24,9 and body weight between 50kg and 120kg Exclusion Criteria: Insufficient linguistic communication Drug abuse or alcohol dependency regular medication except for L-thyroxine or antihistamines known severe eye disease or severely impaired eyesight or hearing a known disease of the cardiovascular system, hypertension higher than 160/90mmHg known pulmonary disease, e.g. bronchial asthma known fractures of the spine or skeletal system of the lower extremity surgery within the last 4 to six months intervention group: fear of heights

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 40 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: 2 groups each undergoing one out of two conditions (high stress (bungee jump) or no stress (control))
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Interventions group: High stress condition (bungee jump)<br>25-30 healthy male volunteers participating in a bungee jump | Behavioral: Bungee jump<br>* Bungee jump from a cran<br>|
| Placebo Comparator: Control group: No stress condition<br>10-15 healthy males serving as a control group without undergoing a stress event/intervention | Behavioral: Bungee jump<br>* Bungee jump from a cran<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Proteomics and autophagy processes | Change in protein levels of autophagy biomarkers (LC3II & p62) of isolated PBMCs (peripheral blood mononuclear cells) by Western Blotting. | change from baseline to the stress intervention and a 5- 7 day follow up | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Proteome patterns | Change in protein levels and protein phosphorylation by untargeted mass spectrometry-based proteomics and phosphoproteomics of isolated PBMCs (peripheral blood mononuclear cells). | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Metabolic processes | Metabolic measurements by mass spectrometry, in order to determine variations in plasma metabolites, including targeted analysis of steroid hormones | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Lipid profiling | Targeted and quantitative analysis by mass spectrometry of change in plasma Lipids. | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Saliva Cortisol Levels | Saliva Cortisol Levels in nmol per Liter (nmol/L) after dexamethasone intake will be evaluated and compared ton the control group | comparison between groups | 
| Sleep Efficiency | Assessment of Sleep Efficiency (total time in bed/time asleep during night) by GenActive Actigraphs | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Overall sleep Quality | Sleep diary to assess overall sleep quality assessed as ratio of the total time spent asleep (in hours) to the total amount of time spent in bed (in hours) per night | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Sleep Quality (PSQI) | Pittsburgh Sleep Quality Index (PSQI): self-report questionnaire to assess sleep quality over a 1-month time interval consisting of 19 individual items. | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Mental well-being (WEMWBS) | Warwick-Edinburgh Mental Well-being Scale (WEMWBS): self-reported 14-item scale to assess Overall mental wellbeing, minimum value 14, maximum value 70, high score indicating high well-being | change from baseline to the stress intervention and a5- 7 day follow up | 
| Resilience behavior (Wagnild &Young) | Resilience scale (Wagnild &Young): self-reported 25-item scale to assess overall resilience, minimum value 25, maximum value 175, high score indicating higher resilience | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Phosphoproteome patterns | Change in protein phosphorylation by untargeted mass spectrometry-based phosphoproteomics of isolated PBMCs (peripheral blood mononuclear cells). | change from baseline to the stress intervention and a 5- 7 day follow up | 
| Ubiquitinome patterns | Change in protein ubiquitination levels by untargeted mass spectrometry-based proteomics of isolated PBMCs (peripheral blood mononuclear cells). | change from baseline to the stress intervention and a 5- 7 day follow up | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Stress, Bungee Jump, Autophagy