Small RNA Profiling in EBC in Athletes

NCT07324252 | Completed

• 2 other identifiers: APK.002.584.2024B.SUB.25.512
• Study Type: observational
• Target: 25
• Locations: 1 country
• Sites: 1

Brief Summary

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.

Conditions

Athlete; RNA Profile; Cardio-pulmonary Function; Exhaled Breath Condensate

Outcome Measures

Primary Outcomes (2)

• smallRNA profile

  From August 2025 to December 2025

• EBC collection

  From January 2025 to August 2025

Study Arms (1)

Exhaled breath condensate donors

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

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

healthy volouteers, amateur athletes (runners) and lung cancer patients with stage IIIb or VI disease

You may qualify if:

• ability to perform CPET
• ability to perform spirometry

You may not qualify if:

• lack of informed consent
• absolute contradictions to CPET or spirometry

Sponsors & Collaborators

• Medical University of Bialystok: lead

Study Sites (1)

Department of Respiratory Physiopathology, Medical University of Bialystok

Bialystok, Podlaskie Voivodeship, 15-540, Poland

Location

Biospecimen

Retention: SAMPLES WITHOUT DNA

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.

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Head of Department of Respiratory Physiopathology

Study Record Dates

• First Submitted: December 23, 2025
• First Posted: January 7, 2026
• Study Start: January 5, 2025
• Primary Completion: August 30, 2025
• Study Completion: December 15, 2025
• Last Updated: February 6, 2026

Record last verified: 2026-02

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP, ICF, CSR
• Time Frame: January 2026 to January 2027
• Access Criteria: researchers and interested parties after an assessment of the data usage description and appropriate bioethical agreement

Locations

PL (1)