Identifying Biological Markers for Altitude Exposure and Use of Recombinant Human Erythropoietin (rHuEPO)

Identification of Biological Markers for Hypoxic Exposure and Use of Recombinant Human Erythropoietin

The project represents a well powered study of the human response to altitude exposure combined with rHuEPO treatment. A total of 20 male and 20 female non-competing athletes will participate. The participants will be exposed to a period of hypobaric hypoxia at 2.320 m for four weeks and a sea-level intervention period of four weeks. The participants will be blinded and randomly allocated to intra venous injection with 20 IU per kg body weight of recombinant human erythropoietin or placebo every second day for the initial three weeks of each intervention. Included subjects will be runners due to the good possibility of recruitment and conduction of training at altitude. Samples are collected weekly for four weeks prior to each intervention period, during the four week intervention and four weeks after. This approach yields a highly valuable biobank for identification of markers sensitive to initiation of rHuEPO injections as well as termination of injections.

Study Overview

• Status: Active, not recruiting
• Conditions: Healthy
• Intervention / Treatment: Other: Sea level; Other: Altitude

Detailed Description

The main hypothesis for this study is that metabolomics and proteomics methods can identify biomarkers that are sensitive to altitude exposure and recombinant human erythropoietin treatment, respectively. Secondarily, the hypothesis is that the identified biomarkers can distinguish between altitude exposure and abuse of rHuEPO.

We will include 20 Danish men and 20 Danish women aged 18-35 years. The inclusion criteria are a maximum oxygen uptake rate (VO2max) > 48 ml / kg / min for men and > 43 ml / kg / min for women. All trial participants must have > 2 years of history in regular exercise including running (> 2 times a week; > 30 min per session) and capable of performing a 5 km run in 22 min for men and 25 min for women at sea level on a level surface in dry conditions and a temperature between 10 and 20 degrees C. Furthermore, the participants must not have experienced running related injuries within the past two years.

The experimental protocol consists of two experimental periods, both of which must be performed by all participants. Both periods contain a baseline period of four weeks followed by four weeks training camp and four weeks follow up. The training camp in one period of experiments is performed at sea level while the training camp in the other period is performed at 2320 m above sea level. Each period is separated by a minimum of two months to ensure that the participants have returned to baseline values. During the two intervention periods, the participants will be transported by air and car to the destination of the respective training camp. It is expected that the participants train for 1-2 hours a day on average and that the time spent for the measurements described is ~ 5-7 hours a week.

To investigate whether the described methods can identify sensitive biomarkers for the physiological response due to rHuEPO treatment, the participants will be treated with rHuEPO.

The distribution of participants allows the following:

• 28 participants receive rHuEPO at sea level, thereby identifying biomarkers sensitive for use of rHuEPO
• 28 participants receive placebo under altitude exposure, whereby biomarkers sensitive to altitude exposure can be identified
• 12 participants receive placebo at sea level, whereby random fluctuations of the identified biomarkers can be determined.
• 12 participants receive rHuEPO under altitude exposure, whereby the identified rhuEPO biomarkers and altitude exposure can be tested for whether they can be used to separate individuals receiving rHuEPO under altitude exposure from individuals receiving placebo under altitude exposure.
• 8 participants receive placebo at sea level and altitude exposure, whereby natural and random individual fluctuations can be determined over a longer period (> 6 months). Since random individual fluctuations can be determined at n = 1, n = 8 will be sufficient to determine this.

• Study Type: Interventional
• Enrollment (Estimated): 40
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Denmark
  • Copenhagen Ø
    • Copenhagen, Copenhagen Ø, Denmark, 2200
      • University of Copenhagen

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 35 years (Adult)
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Male: VO2max > 48 ml/min/kg bodyweight and 5km run < 22 min, Female: VO2max > 43ml/min/kg bodyweight and 5km run < 25 min A minimum of 2 years injury running training

Exclusion Criteria:

• Age, insufficient fitness or injury

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Other
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Quadruple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Sea level; Sea level training camp	Other: Sea level; Sea level exposure / sea level exposure combined with microdoses of rHuEPO
Experimental: Altitude; Altitude training camp	Other: Altitude; Altitude exposure / altitude exposure Iron metabolism

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Identification of metabolites in urine for rHuEPO and altitude exposure	Urine is analyzed by metabolomics and the metabolites for rHuEPO and altitude are verified using an exploratory design. More specifically, the aim is to identify the resulting metabolites from altitude exposure and treatment with rHuEPO as these are not yet known In this WADA-supported research project, the aim is to identify high sensitivity biomarkers for altitude exposure through a metabolomics methods, as 'omics' methods have previously shown promising results in other exposure contexts. However, being the first of its kind in antidoping the resulting metabolites from rHuEPO misuse and altitude have not yet been identified. Thus, an exact description of these is not possible	5 years
Identification of human metabolites in serum for rHuEPO and altitude exposure	Human serum is analyzed by metabolomics and the metabolites for rHuEPO and altitude are verified using an exploratory design.	5 years

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Reticulocyte percentage	Reticulocyte percentage, will be measured to determine variation as a result of either 12 weeks exposure to altitude or sea-level (24 weeks in total).	Up to 24 weeks
Hemoglobin concentration	[hemoglobin] will be measured to determine variation as a result of either 12 weeks exposure to altitude or sea-level.	Up to 24 weeks
OFF-score	OFF-score (computed from reticulocyte perscentage and hemoglobin concentration), will be measured to determine variation as a result of either 12 weeks exposure to altitude or sea-level.	Up to 24 weeks
Maximal oxygen uptake (VO2max)	VO2max (ml/min) measured by incremental work load on a treadmill	Up to 24 weeks
Iron metabolism	Markers of iron metabolism such as hepcidin and erythroferrone are collected analyzed for evaluation of iron metabolism during and after a training camp at 2300m above sea level	Up to 24 weeks
Work economy	Running efficiency (%) measured by energy expenditure on a treadmill	Up to 24 weeks
Sustainable workload	Lactate threshold (mmol/L) measured on a treadmill	Up to 24 weeks
Running performance	5000m time trial performance (time) measured on a 400m running track	Up to 24 weeks
Muscle oxygenation	Muscle oxygenation measured by near-infrared spectroscopy during running on a treadmill	Up to 24 weeks
Total hemoglobin mass	Total hemoglobin mass will be measured by two-minute carbon-monoxide re-breathing to determine variation as a result of either 12 weeks exposure to altitude or sea-level.	Up to 24 weeks

Collaborators and Investigators

• Sponsor: University of Copenhagen
• Collaborators: Oslo University Hospital; Universidad de Granada; German Sport University, Cologne; World Anti-Doping Agency
• Investigators: Principal Investigator: Jacob Bejder, Msc, Universuty of Copenhagen

Publications and helpful links

General Publications

• Arrebola-Moreno AL, Casuso RA, Bejder J, Bonne TC, Breenfeldt Andersen A, Aragon-Vela J, Nordsborg NB, Huertas JR. Does Hypoxia and Stress Erythropoiesis Compromise Cardiac Function in Healthy Adults? A Randomized Trial. Sports Med Open. 2022 Nov 5;8(1):137. doi: 10.1186/s40798-022-00531-x.

Study record dates

Study Major Dates

• Study Start (Actual): March 1, 2018
• Primary Completion (Actual): June 26, 2019
• Study Completion (Estimated): May 30, 2025

Study Registration Dates

• First Submitted: May 7, 2018
• First Submitted That Met QC Criteria: January 9, 2020
• First Posted (Actual): January 14, 2020

Study Record Updates

• Last Update Posted (Actual): November 29, 2023
• Last Update Submitted That Met QC Criteria: November 28, 2023
• Last Verified: November 1, 2023

More Information

Terms related to this study

• Other Study ID Numbers: H-17036662

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: Fully anonymous data will be shared with other researchers.

Drug and device information, study documents

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