A Pilot Study of the VosCryoTM Breathalyser in Lung Cancer

Overview

CryoPilot is a single-centre pilot study being conducted at Weston Park Cancer Centre in Sheffield, UK. The goal of this pilot study is to determine whether small-volume exhaled breath condensate sample (EBC) obtained via a VosCryo™ breath sample collection device can provide quantifiable and analysable cellular material, including epithelial cells, leukocytes, or exfoliated tumour cells, in patients with late-stage lung cancer. The main question it aims to answer is whether a breath sample can provide the necessary cellular information that could eventually be used for diagnostic purposes. The secondary objective is to compare the presence and quality of such material with that obtained from healthy relatives who will act as a low-risk control group, to assess whether EBC-based analysis may differentiate between cancerous and non-cancerous states. As such this pilot study will have two arms: * Experimental arm: 12 patients with late-stage lung cancer * Control arm: 12 low-risk relatives of patients with late-stage lung cancer Participants will be approached during their visit to Weston Park Hospital and asked to provide informed consent, followed by provision of a breath sample.

Detailed description

Lung cancer remains one of the leading causes of cancer-related mortality worldwide, with late-stage diagnosis contributing substantially to poor prognosis (Bray et al., 2024). Despite advancements in imaging and molecular diagnostics, early detection strategies remain inadequate for population-wide screening, particularly in resource-limited settings. There is therefore an urgent need for novel, non-invasive, and cost-effective diagnostic approaches suitable for use in at-risk populations, such as individuals with a family history of lung cancer or those with a history of significant exposure to tobacco smoke.

Exhaled breath condensate (EBC) offers a promising, non-invasive source of biomarkers for respiratory diseases including lung cancer (Carpagnano et al., 2005; Palomba et al., 2023). EBC is a biological fluid formed by the cooling of exhaled breath and contains aerosolised droplets from the respiratory tract, including proteins, nucleic acids, volatile organic compounds, and, occasionally, cellular material (Horvath et al., 2005; Mutlu et al., 2001). These components may reflect pathological processes occurring in the lower airways and alveoli, thus offering a window into the disease state without the risks associated with bronchoscopy or biopsy.

Recent technological developments have enabled the reproducible collection of small volumes of EBC in a clinically practical timeframe. The VosCryo™ device, is a commercially available disposable device that collects approximately 100-200 µL of condensate in about one minute. The VosCryo™ which we will use, is a fast, non-invasive, and reliable EBC collector, optimized for small-volume sampling (50-200 µL/min).

Primarily the study aims to determine whether small-volume EBC obtained via VosCryo™ from late-stage lung cancer patients contains quantifiable and analysable cellular material, including epithelial cells, leukocytes, or exfoliated tumour cells. This will be determined from the proportion of participants with lung cancer with analysable material within their exhaled breath condensate. The study also aims to compare the presence and quality of such material with that obtained from healthy relatives (acting as a low-risk control group), to assess whether EBC-based analysis may differentiate between cancer and non-cancer states, and therefore will recruit from two cohorts - patients with late stage lung cancer and relatives of late-stage cancer patients with no evidence/low risk of cancer. This exploratory pilot study will inform the viability of future development of EBC as a low-cost, non-invasive diagnostic in patients with advanced and early-stage lung cancer, and as a potential screening tool for use in at-risk populations, and may lead to larger scale studies.

This will be a single-centre pilot study run at Weston Park Cancer Centre in Sheffield, United Kingdom, with patients and relatives recruited by clinical/medical oncologists and clinical trials assistants during their routine oncology clinic appointments. Patients attending the lung cancer clinic without an accompanying relative can be approached for study participation. Consenting patients with an accompanying relative who declines study participation can be included, as can consenting relatives with a patient who declines.

With patient consent, the study will collect a range of data at the single baseline visit for descriptive statistical analysis, including:

* Sex. * Age. * Smoking status - never smoker, ex-smoker (not smoked \>12 months), recent ex-smoker (smoked ≤12 months), current smoker (smoked within past 14 days), vapes. * TNM tumour stage 9th edition (where appropriate) from most recent staging scans performed as part of NHS standard of care. * Tumour histology (where appropriate) including histological subtype, immunohistochemistry profile, mutation profile and PD-L1 status from results reported as part of NHS standard of care investigations. * History of any concurrent pulmonary disease (e.g. emphysema, interstitial lung disease)

EBC Research Samples will be analysed using immunocytochemistry, indirect RNA extraction, cDNA synthesis and qPCR-based analysis of genes expressed in epithelial and immune cell types. The samples will also be stored for the duration of the project (18 months), and may be re-analysed in future projects using techniques such as ELISA, fluorescence microscopy or mass spectrometry.

It is estimated that participant recruitment and sample collection will be completed over a 6-month period, with subsequent sample and data analysis completing over an additional 12-month period.

Interventions

• Other Expiratory Breath Condensate Collection
  Collection of Expiratory Breath Condensate using the VosCryo device

Primary outcome measures

• Availability of Analysable Sample Material [Time frame: At Collection of EBC Sample]

Secondary outcome measures (1)

• Cross-Cohort Sample Discrepancy [Time frame: Variation in gene expression identified at end of study, from samples collected at baseline visit.]

Eligibility criteria

Inclusion criteria

• Age ≥16 years old
• Ability to provide written informed consent
• Either a history of advanced non-small cell or small cell lung cancer with intra-thoracic disease (pulmonary primary or pulmonary metastases), previously untreated by systemic anti-cancer treatment or radiotherapy

• Patients with previously treated early-stage lung cancer are permitted
• Patients with advanced disease with extra-thoracic disease previously treated by local therapy (e.g., palliative radiotherapy to a bone metastasis, or post brain metastectomy), which in the opinion of the Principal Investigator is unlikely to impact the quality of the EBC research sample are eligible
• OR an attending relative or friend with no history of lung cancer or any other primary cancer, except for curatively treated non-melanomatous skin cancer or carcinoma in situ, unless the participant has been free of malignancy and off treatment for a period of at least 2 years prior to study enrolment
• No intercurrent upper or lower respiratory tract infection
• Able to complete the EBC sample collection

Exclusion criteria

• Participants with lung metastases from non-lung primary tumours
• Participants unable to consent or unable to comply with the requirements for EBC sample collection

Criteria are shown verbatim from the registry (in English). Final eligibility is always assessed by the study center.

Healthy volunteers: Yes

Study design

Observational model

Case-control

Study locations

United Kingdom · 1 center

• Weston Park Cancer Centre — Sheffield

Publications

• Phillips M, Cataneo RN, Cummin AR, Gagliardi AJ, Gleeson K, Greenberg J, Maxfield RA, Rom WN. Detection of lung cancer with volatile markers in the breath. Chest. 2003 Jun;123(6):2115-23. doi: 10.1378/chest.123.6.2115. PMID 12796197
• Palomba L, Paris D, Tramice A, Ambrosino P, Maniscalco M, Motta A. Detection of Cells in Exhaled Breath Condensate Holds Potential for Pathophysiological Insights in Pulmonary Diseases. Am J Respir Cell Mol Biol. 2023 Jul;69(1):113-115. doi: 10.1165/rcmb.2023-0022LE. No abstract available. PMID 37387613
• Mutlu GM, Garey KW, Robbins RA, Danziger LH, Rubinstein I. Collection and analysis of exhaled breath condensate in humans. Am J Respir Crit Care Med. 2001 Sep 1;164(5):731-7. doi: 10.1164/ajrccm.164.5.2101032. No abstract available. PMID 11549524
• Kharitonov SA, Barnes PJ. Exhaled biomarkers. Chest. 2006 Nov;130(5):1541-6. doi: 10.1378/chest.130.5.1541. PMID 17099035
• Horvath I, Hunt J, Barnes PJ, Alving K, Antczak A, Baraldi E, Becher G, van Beurden WJ, Corradi M, Dekhuijzen R, Dweik RA, Dwyer T, Effros R, Erzurum S, Gaston B, Gessner C, Greening A, Ho LP, Hohlfeld J, Jobsis Q, Laskowski D, Loukides S, Marlin D, Montuschi P, Olin AC, Redington AE, Reinhold P, van Rensen EL, Rubinstein I, Silkoff P, Toren K, Vass G, Vogelberg C, Wirtz H; ATS/ERS Task Force on E PMID 16135737
• Department of Health and Social Care. (2017). UK Policy Framework for Health and Social Care Research. (accessed at https://www.hra.nhs.uk/planning-and-improving-research/policies-standards- legislation/uk-policy-framework-health-social-care-research/)
• Carpagnano GE, Foschino-Barbaro MP, Mule G, Resta O, Tommasi S, Mangia A, Carpagnano F, Stea G, Susca A, Di Gioia G, De Lena M, Paradiso A. 3p microsatellite alterations in exhaled breath condensate from patients with non-small cell lung cancer. Am J Respir Crit Care Med. 2005 Sep 15;172(6):738-44. doi: 10.1164/rccm.200503-439OC. Epub 2005 Jun 9. PMID 15947287
• Bray F, Laversanne M, Sung H, Ferlay J, Siegel RL, Soerjomataram I, Jemal A. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024 May-Jun;74(3):229-263. doi: 10.3322/caac.21834. Epub 2024 Apr 4. PMID 38572751

Identifiers

NCT: NCT07689968 · STH23461 · IRAS

Primary sources (government registries)

• ClinicalTrials.gov (US NIH)

View this study on ClinicalTrials.gov ↗