Hybrid-sensor Breath Analysis for Colorectal Cancer Screening

Study Overview
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Brief Summary
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The aim of this project is to promote the breath volatile marker concept for colorectal cancer (CRC) screening by advancing developing the application of a novel hybrid analyzer for the purpose. The hybrid analyzer concept is expected to benefit of combining metal-oxide (MOX) and infrared spectrum (IR) sensor acquired data. The current study will be the first globally to address this concept in CRC detection. In addition, traditional methods, in particular, gas chromatography coupled to mass spectrometry (GC-MS) will be used to address the biological relevance of the VOCs emission from cancer tissue and will assist in further advances of the hybrid-sensing approach.

Detailed Description
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For addressing the aims of the project, four specific research objectives have been set: To identify cancer-related VOCs emitted by the CRC tissue via the comparison of VOCs emitted from cancer tissue with VOCs emitted by non-cancerous tissue (ex vivo surgery material) by GC-MS. To identify the VOCs differentiating human breath from CRC patients and controls (by GC-MS) as well as compare the chemical signature of CRC patients' breath to the chemical signature of cancer tissue. To evaluate the performance of the set of sensors in the hybrid analyzer and the performance of particular sensors for detecting CRC; to develop and validate a mathematical model for CRC detection. To validate the hybrid analyzer in real-life CRC screening settings, i.e. versus the generally accepted CRC screening approach of faecal occult blood detection. To compare faecal microbiome between CRC group and control. The scientific results to be obtained during the current project are expected to elucidate the origin and metabolism of volatile biomarkers of CRC. This achievement, in turn, will facilitate the implementation of a new screening test based on the newly developed hybrid analyser into medical practice. Identification of the VOCs patterns by the sensor array for CRC patients when compared to controls. Addressing these objectives will allow an in-depth understanding of the physiological background for exhaled VOCs in CRC patients and facilitate the development of technologies able to identify the disease and its precursors from an exhaled breath sample.

Official Title
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Hybrid-sensor Breath Analysis for Colorectal Cancer Screening (HYCOR)

Conditions
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Colorectal Cancer, Polyp of Colon

Intervention / Treatment
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* Procedure: Identification of specific VOCs in CRC tissue surgery material
* Other: Secondary validation study in general CRC screening settings
* Device: Breath sampling for VOC detection
* Other: Blood sample collection
* Diagnostic Test: Microbiota testing
* Diagnostic Test: Colonoscopy


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Adult individuals (>18 years of age) Having signed the consent form Willingness to collaborate Able to provide a breath sample For the cancer group: colorectal adenocarcinoma has to be documented histologically (histological diagnosis following gastric surgery is also accepted) or patients being confirmed adenocarcinoma during the course of the study. For the non-cancer group: control group - any patient who have medical indications for a colonoscopy Exclusion Criteria: The patient has not signed the consent form Patients who have had a complete bowel cleansing Other active malignancies Neoadjuvant chemotherapy, radiation therapy is currently underway Acute conditions (emergency surgery for the patient) Small bowel resection in the past Terminal renal failure (Chronic renal failure stage 4) Type I diabetes Bronchial asthma (active)

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Colorectal cancer patients<br>Patients with histologically confirmed colorectal cancer (adenocarcinoma) | Device: Breath sampling for VOC detection<br>* Breath sampling will be performed by using a hybrid sensor device and or GC-MS analysis (by collecting breath samples in adsorbent tubes). Strict requirements for subjects will be imposed prior to the breath sampling to standardise the breath sampling and to limit the influence of confounding factors.<br>Other: Blood sample collection<br>* Serum, plasma sampling for group description and stratification.<br>Diagnostic Test: Microbiota testing<br>* Faecal samples for microbiota testing.<br>|
| Control group patients without colorectal cancer<br>Patients without colorectal malignant disease according to data obtained in colonoscopy | Device: Breath sampling for VOC detection<br>* Breath sampling will be performed by using a hybrid sensor device and or GC-MS analysis (by collecting breath samples in adsorbent tubes). Strict requirements for subjects will be imposed prior to the breath sampling to standardise the breath sampling and to limit the influence of confounding factors.<br>Other: Blood sample collection<br>* Serum, plasma sampling for group description and stratification.<br>Diagnostic Test: Microbiota testing<br>* Faecal samples for microbiota testing.<br>Diagnostic Test: Colonoscopy<br>* Colonoscopy will be used only according to the clinical indications.<br>|
| Average risk population<br>Average risk population of both genders aged 40-64 at the time of inclusion lacking alarm symptoms for gastrointestinal cancer | Other: Secondary validation study in general CRC screening settings<br>* Altogether at least 1000 individuals relatively healthy 40-64 years old population-based collected individuals will get recruited. Breath samples will be collected by asking the study subjects to breath into hybrid breath analyser. To exclude significant colorectal lesions, laboratory-based FIT testing will be offered to the population cohort group for faecal occult blood in faeces. Serum and plasma samples will also be obtained to have them available if additional testing will be required. Individuals with a FIT test value over the cut-off value (>10 microg/g faeces) will be invited to colonoscopy. The data analysis procedures and classification models will be tested in this general population and cross-checked against FIT and colonoscopy results.<br>Other: Blood sample collection<br>* Serum, plasma sampling for group description and stratification.<br>Diagnostic Test: Microbiota testing<br>* Faecal samples for microbiota testing.<br>Diagnostic Test: Colonoscopy<br>* Colonoscopy will be used only according to the clinical indications.<br>|
| Colorectal cancer patients undergoing surgery<br>Patients with histologically confirmed colorectal cancer (adenocarcinoma) planned for surgical management | Procedure: Identification of specific VOCs in CRC tissue surgery material<br>* Paired tissue samples will be taken during surgery for CRC. Tissue material from the same patient will be obtained from the cancerous tissue as well as from normal resected material without malignant infiltration. Minimum of 100 mg of each tissue per sample will be obtained. To compare the emission of VOCs in the CRC tissue surgery material to the emissions from normal tissue by GC-MS in a reasonable number of cancer cases.<br>Other: Blood sample collection<br>* Serum, plasma sampling for group description and stratification.<br>Diagnostic Test: Microbiota testing<br>* Faecal samples for microbiota testing.<br>|
| Patients with polyps undergoing polypectomy<br>Patients with colon polyps that will perform polypectomy | Device: Breath sampling for VOC detection<br>* Breath sampling will be performed by using a hybrid sensor device and or GC-MS analysis (by collecting breath samples in adsorbent tubes). Strict requirements for subjects will be imposed prior to the breath sampling to standardise the breath sampling and to limit the influence of confounding factors.<br>Other: Blood sample collection<br>* Serum, plasma sampling for group description and stratification.<br>Diagnostic Test: Microbiota testing<br>* Faecal samples for microbiota testing.<br>Diagnostic Test: Colonoscopy<br>* Colonoscopy will be used only according to the clinical indications.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Characteristic VOC pattern identification for colorectal cancer detection | The characteristic VOC pattern based on sensor analysis and its performance indicators will be detected. | 2 years following initiation of patient recruitment | 
| Specific chemistry identification in the exhaled breath | Identification of specific chemistries (GC-MS analysis) originating from colorectal cancer. Volatiles will be separated using an Rt-Q-BOND column working in a constant flow of helium. The column temperature program will be optimized toward detection of observed volatiles. The SCAN, will be used for the untargeted analysis and identification of compounds of breath samples as well as for the quantification of more abundant species. Peak integration will be based on extracted ion chromatograms. The identification of compounds will be performed in two steps. The peak spectrum will be checked against the NIST mass spectral library. The NIST identification will be confirmed by comparing the respective retention times with retention times obtained on the basis of standard mixtures prepared from pure compounds. Whenever possible the VOC emission will be quantified using calibration mixtures prepared from pure liquid or gaseous substances. | 2 years following initiation of patient recruitment | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Identification of the best-performing sensors | Decision on the optimal set of breath sensors that potentially will be included in a sensor analyser for CRC detection. Comparative analysis between the performance of different sensor performance in target disease identification. | 3 years following initiation of patient recruitment | 
| Gut microbiota analysis in relation to breath VOCs | Analysis of the role of faecal microbiota in the origin of VOCs in the exhaled breath. | 3 years following initiation of patient recruitment | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Volatile organic compounds, Colorectal cancer, Breath testing