Small RNA Profiling in EBC in Athletes

NCT ID: NCT07324252

Last Updated: 2026-01-07

Study Results

Results pending

The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.

Basic Information

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Recruitment Status

COMPLETED

Total Enrollment

25 participants

Study Classification

OBSERVATIONAL

Study Start Date

2025-01-05

Study Completion Date

2025-12-15

Brief Summary

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Background: Exhaled breath condensate (EBC) is a non-invasively collected biofluid containing volatile and non-volatile compounds, including small RNAs. Small RNA profiling in EBC appears to be a promising approach for identifying and developing minimally invasive biomarkers for lung diseases; however, methodological evidence remains limited and heterogeneous.

Objectives: To evaluate the technical feasibility of small RNA extraction and next-generation sequencing (NGS) from EBC and to characterise small RNA profiles at rest and after maximal exercise, using EBC as a model matrix for dynamic physiological changes.

Methods: In a pilot study conducted as part of statutory research at the Medical University of Białystok (B.SUB.25.512 and B.SUB.25.529), we analysed 22 EBC samples collected with the RTube device from 11 healthy volunteers and 3 patients with lung cancer and co-existing COPD (GOLD 3-4), at rest and after standardised maximal exercise confirmed by cardiopulmonary exercise testing (CPET). Total RNA was extracted using a commercial high-throughput RNA isolation kit designed for biological samples with low nucleic acid content, the miRNeasy Serum/Plasma Advanced Kit (Qiagen), according to the manufacturer's protocol with modifications. Small RNA libraries were generated using the high-sensitivity, low-input TrueQuant Small RNA-Seq Library Preparation Kit (GenXPro GmbH) with Unique Molecular Identifiers (UMIs) and sequenced on an Illumina platform. Reads were processed with Cutadapt and FastQC, mapped iteratively to the hg38 (Homo sapiens) reference genome using Bowtie2, aligned against an array of RNA databases, including miRNA (miRBase), tRNA (GtRNAdb), piRNA (piRNAdb), ncRNA (ENSEMBL), and cDNA (ENSEMBL) and quantified with HTSeq. Differential expression analysis (DEA) and principal component analysis (PCA) were performed with DESeq2.

Detailed Description

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Conditions

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Exhaled Breath Condensate Athlete RNA Profile Cardio-pulmonary Function

Study Design

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Observational Model Type

COHORT

Study Time Perspective

PROSPECTIVE

Study Groups

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Exhaled breath condensate donors

athletes (22) and lung cancer (3) donors of EBC

No interventions assigned to this group

Eligibility Criteria

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Inclusion Criteria

* ability to perform CPET
* ability to perform spirometry

Exclusion Criteria

* lack of informed consent
* absolute contradictions to CPET or spirometry
Minimum Eligible Age

18 Years

Eligible Sex

ALL

Accepts Healthy Volunteers

Yes

Sponsors

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Medical University of Bialystok

OTHER

Sponsor Role lead

Responsible Party

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Lukasz Minarowski

MD PhD

Responsibility Role PRINCIPAL_INVESTIGATOR

Locations

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Department of Respiratory Physiopathology, Medical University of Bialystok

Bialystok, Podlaskie Voivodeship, Poland

Site Status

Countries

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Poland

Other Identifiers

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APK.002.584.2024

Identifier Type: -

Identifier Source: org_study_id

B.SUB.25.512

Identifier Type: OTHER

Identifier Source: secondary_id

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