Multi-level Molecular Profiling of Peak Performance in Endurance Sports (MSSA)

A Pilot-study on Multi-level Molecular Profiling of Peak Performance in Endurance Sports

Physical activity triggers complex molecular responses, including changes in immune-, stress-, and metabolic pathways. For example, autophagy is essential for energy and cellular homeostasis through protein catabolism, and dysregulation results in compromised proteostasis, reduced exercise performance, and excessive secretion of signaling molecules and inflammatory proteins. However, previous research has been limited by the extend of molecules measured and biological processes covered. A better understanding of these processes through multi-omic analysis can improve knowledge of molecular changes in response to exercise. The main purpose of the investigators study is to analyze the effects of acute exercise in correlation to autophagy and other signaling cascades. Specifically, the investigators plan to perform multi-level molecular profiling in a cohort of healthy male elite cyclists and male and female recreational athletes, before, during, and after a bicycle ergometer test. The results will be compared to a control cohort without intervention.

Study Overview

• Status: Recruiting
• Conditions: Healthy; Exercise; Physical Fitness; Exercise Test
• Intervention / Treatment: Diagnostic test: Performance testing

Detailed Description

This is a non-randomized controlled trial performed at the Paracelsus Medical University, Salzburg, Austria. The study will recruit 80 healthy men and women. Subjects who meet the inclusion criteria will be allocated to four arms (n = 20 in all groups): 1. elite cyclists, 2. male recreational athletes, 3. female recreational athletes, 4. male control group.

After overnight fasting and medical check-up, groups 1-3 will undergo a bicycle ergometer-based exercise protocol designed to span low (aerobic) to severe (anaerobic) domains of exercise. The protocol consists of a 15 min aerobic warm-up phase followed by a ramp-bicycle ergometer protocol.

During exercise, performance-relevant data will be continuously monitored. Venous blood specimens will be collected before exercise (baseline), at the end of the warm-up as well as 2 min, 10 min, and 30 min in recovery.

• Study Type: Interventional
• Enrollment (Anticipated): 80
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Jens Stepan, MD, PhD; Phone Number: +43(0)5725555625; Email: j.stepan@salk.at
• Study Contact Backup: Name: Nils Gassen, PhD; Phone Number: +49(0)22828715793; Email: nils.gassen@ukbonn.de

Study Locations

• Austria
  • Salzburg, Austria, 5020
    • Recruiting
    • Paracelsus Medical University
    • Contact: Jens Stepan, MD, PhD; Phone Number: +43(0)5725555625; Email: j.stepan@salk.at

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 40 years (Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• All arms: ability to give written informed consent
• Arms 1- 3: inconspicuous medical examination (medical history, resting ECG, echocardiography)
• Elite athletes (arm1, cyclists): maximum oxygen uptake > 65 ml/kg/KG
• Elite athletes (arm1, cyclists): participation in cycling competitions on a regular basis
• Recreational athletes (arms 2 and 3): maximum oxygen uptake < 65 ml/kg/KG for male subjects and < 55 ml/kg/KG for female subjects

Exclusion Criteria:

• All arms: unable to communicate adequately by language
• All arms: regular use of prescription drugs other than thyroxine or antihistamines
• All arms: alcohol consumption as equivalent doses averaging more than 40 g of pure alcohol per day
• All arms: use of illicit drugs
• All arms: known diseases of the cardiovascular system
• All arms: arterial hypertension over 160/90 mmHg at rest
• All arms: known pulmonary diseases, especially bronchial asthma
• All arms: surgery less than 4-6 months ago.
• All arms: abnormalities in the medical examination (medical history, resting ECG, echocardiography)
• Arms 1-3: orthopaedic diseases that preclude maximum exercise on a bicycle ergometer
• Elite athletes (arm1, cyclists): maximum oxygen uptake < 65 ml/kg/KG
• Recreational athletes (arms 2 and 3): maximum oxygen uptake > 65 ml/kg/KG for male subjects and > 55 ml/kg/KG for female subjects
• Arm 3: positive urine ß-HCG

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Elite cyclists, male; Bicycle ergometer-based exercise testing. 15 min aerobic warm-up phase (2 W/kg) followed by a standardized but individualized ramp-bicycle ergometer protocol to reach maximal exercise capacity after 8 - 12 minutes. The exclusively aerobic energy supply during warm-up will be assessed by constant respiratory quotient and constant arterial lactate concentration (<0.5 mmol in the last 5 min). During exercise, respiratory gas exchange, heart rate, blood pressure, ECG and ratings of perceived exertion will be continuously monitored. Venous blood specimens will be collected before exercise (baseline), at the end of the warm-up as well as 2 min, 10 min, and 30 min in recovery.	Diagnostic test: Performance testing; Performance testing on a bicycle ergometer.
Experimental: Recreational athletes, male; Bicycle ergometer-based exercise testing. 15 min aerobic warm-up phase (1 W/kg) followed by a standardized but individualized ramp-bicycle ergometer protocol to reach maximal exercise capacity after 8 - 12 minutes. The exclusively aerobic energy supply during warm-up will be assessed by constant respiratory quotient and constant arterial lactate concentration (<0.5 mmol in the last 5 min). During exercise, respiratory gas exchange, heart rate, blood pressure, ECG and ratings of perceived exertion will be continuously monitored. Venous blood specimens will be collected before exercise (baseline), at the end of the warm-up as well as 2 min, 10 min, and 30 min in recovery.	Diagnostic test: Performance testing; Performance testing on a bicycle ergometer.
Experimental: Recreational athletes, female; Bicycle ergometer-based exercise testing. 15 min aerobic warm-up phase (1 W/kg) followed by a standardized but individualized ramp-bicycle ergometer protocol to reach maximal exercise capacity after 8 - 12 minutes. The exclusively aerobic energy supply during warm-up will be assessed by constant respiratory quotient and constant arterial lactate concentration (<0.5 mmol in the last 5 min). During exercise, respiratory gas exchange, heart rate, blood pressure, ECG and ratings of perceived exertion will be continuously monitored. Venous blood specimens will be collected before exercise (baseline), at the end of the warm-up as well as 2 min, 10 min, and 30 min in recovery.	Diagnostic test: Performance testing; Performance testing on a bicycle ergometer.
No Intervention: Control, male; Venous blood specimens will be collected at the same time points in the absence of exercise.

What is the study measuring?

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.	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Salivary cortisol levels	Salivary cortisol levels in nmol per Liter (nmol/L) after will be evaluated and compared to the control group	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Proteome patterns	Change in protein levels and protein phosphorylation by untargeted mass spectrometry-based proteomics and phosphoproteomics of isolated PBMCs (peripheral blood mononuclear cells).	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Metabolic processes	Targeted and quantitative analysis by mass spectrometry of change in metabolites of Plasma.	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Lipid profiling	Targeted and quantitative analysis by mass spectrometry of change in plasma Lipids.	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Phosphoproteome patterns	Change in protein phosphorylation by untargeted mass spectrometry-based phosphoproteomics of isolated PBMCs (peripheral blood mononuclear cells).	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Ubiquitinome patterns	Change in protein ubiquitination levels by untargeted mass spectrometry-based proteomics of isolated PBMCs (peripheral blood mononuclear cells).	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Epigenetic patterns	Evaluate epigentic methylation patterns through blood based epigenome analysis	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.
Exosomal protein patterns	Evaluate exosomal protein content through mass spectrometry based analysis	Before the exercise test (baseline), at the end of the 15 minute warm-up phase and 2 minutes, 10 minutes and 30 minutes after termination of the exercise test.

Collaborators and Investigators

• Sponsor: Paracelsus Medical University
• Collaborators: Department of Psychiatry University of Bonn
• Investigators: Principal Investigator: Jens Stepan, MD, PhD, Department of Obstetrics and Gynecology, University Hospital Salzburg, Austria

Publications and helpful links

General Publications

• Hasin Y, Seldin M, Lusis A. Multi-omics approaches to disease. Genome Biol. 2017 May 5;18(1):83. doi: 10.1186/s13059-017-1215-1.
• Nicora G, Vitali F, Dagliati A, Geifman N, Bellazzi R. Integrated Multi-Omics Analyses in Oncology: A Review of Machine Learning Methods and Tools. Front Oncol. 2020 Jun 30;10:1030. doi: 10.3389/fonc.2020.01030. eCollection 2020.
• Contrepois K, Wu S, Moneghetti KJ, Hornburg D, Ahadi S, Tsai MS, Metwally AA, Wei E, Lee-McMullen B, Quijada JV, Chen S, Christle JW, Ellenberger M, Balliu B, Taylor S, Durrant MG, Knowles DA, Choudhry H, Ashland M, Bahmani A, Enslen B, Amsallem M, Kobayashi Y, Avina M, Perelman D, Schussler-Fiorenza Rose SM, Zhou W, Ashley EA, Montgomery SB, Chaib H, Haddad F, Snyder MP. Molecular Choreography of Acute Exercise. Cell. 2020 May 28;181(5):1112-1130.e16. doi: 10.1016/j.cell.2020.04.043.
• Mukherjee K, Edgett BA, Burrows HW, Castro C, Griffin JL, Schwertani AG, Gurd BJ, Funk CD. Whole blood transcriptomics and urinary metabolomics to define adaptive biochemical pathways of high-intensity exercise in 50-60 year old masters athletes. PLoS One. 2014 Mar 18;9(3):e92031. doi: 10.1371/journal.pone.0092031. eCollection 2014.
• De Pauw K, Roelands B, Cheung SS, de Geus B, Rietjens G, Meeusen R. Guidelines to classify subject groups in sport-science research. Int J Sports Physiol Perform. 2013 Mar;8(2):111-22. doi: 10.1123/ijspp.8.2.111.
• Martin DT, McLean B, Trewin C, Lee H, Victor J, Hahn AG. Physiological characteristics of nationally competitive female road cyclists and demands of competition. Sports Med. 2001;31(7):469-77. doi: 10.2165/00007256-200131070-00002.
• Hoffman NJ. Omics and Exercise: Global Approaches for Mapping Exercise Biological Networks. Cold Spring Harb Perspect Med. 2017 Oct 3;7(10):a029884. doi: 10.1101/cshperspect.a029884.

Study record dates

Study Major Dates

• Study Start (Actual): October 15, 2021
• Primary Completion (Anticipated): October 15, 2022
• Study Completion (Anticipated): October 15, 2022

Study Registration Dates

• First Submitted: April 25, 2022
• First Submitted That Met QC Criteria: April 28, 2022
• First Posted (Actual): May 4, 2022

Study Record Updates

• Last Update Posted (Actual): May 4, 2022
• Last Update Submitted That Met QC Criteria: April 28, 2022
• Last Verified: April 1, 2022

More Information

Terms related to this study

• Keywords: Autophagy; Exercise Test; Multi-omic; Fitness level
• Other Study ID Numbers: jestep032022

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

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