New Non-invasive Biomarkers of Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD)

Completed

• Conditions: Healthy; Simple Steatosis (SS); Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD); Metabolic Dysfunction-associated Steatohepatitis (MASH)

• Registration Number: NCT06647095
• Lead Sponsor: General University Hospital, Prague

Brief Summary

The goal of this observational study is to learn if some components of blood or exhaled breath can diagnose people having more fat in their livers than is normal, because of their poorer metabolic health (for example, because of obesity and diabetes). The main questions it aims to answer are:

1. Can a method find participants with higher liver fat than healthy participants?

2. Can a method find participants in whom higher liver fat was a cause of liver inflammation or stiffness?

Participants will:

* fast overnight

* have a routine blood draw

* easily exhale a few times into a special device or a plastic bag and fill in a short dietary questionnaire (if participating in a breath test)

* optionally swallow capsules with an orange peel extract and fish oil before exhaling, which can help get better results from breath (capsules will be medically safe and approved)

Detailed Description

Patients with MASLD and healthy volunteers will be offered an observational study evaluating the diagnostic role of new non-invasive experimental methods (serum bile acids, breath VOC, and plasmatic spectroscopic patterns) assessing the presence and severity of liver fibrosis and steatosis.

First, this study aims to differentiate patients with metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis (MASH) from healthy controls (or simple steatosis) using three experimental methods.

Second, the investigators aim to stratify patients with MASLD according to the presence/grade of steatosis and fibrosis (any, significant F2-3, advanced F3, cirrhosis F4) using the same methods.

Experimental methods tested in this study include:

1. analyzing the fasting spectrum of serum bile acids using liquid chromatography-mass spectrometry

2. analyzing the disease-specific spectroscopy patterns given by vibrational and chiroptical spectroscopy of blood plasma

3. analyzing trace concentrations of volatile organic compounds (VOC) in exhaled human breath using the Selected ion flow tube mass spectrometry. This contains the so-called stress test to monitor breath VOC (d-Limonene and triethylamine) after their regulated ingestion in the form of capsules

These parameters may eventually be combined with anthropometric and laboratory parameters (such as age or BMI).

Clinical examinations, blood sampling, ultrasound examinations of the liver, and liver elastography will be performed as part of routine care.

The results of the blood parameters can be retrospectively evaluated.

Approximately 60-80 participants in total were anticipated for each method. For patients who participate in breath tests, adequate insurance must be guaranteed.

Statistical processing: individual parameters will be evaluated in an exploratory and a validation group or by the PLS-DA algorithm with repeated cross-validation. A difference of p \<0.05 will be considered a statistically significant change.

Study Timeline

• Study Start: 2021-09-01
• Primary Completion: 2022-09-01
• Study Completion: 2024-09-01
• Status Verified: 2024-10
• Study First Submitted: 2024-10-15
• Study First Submitted QC: 2024-10-15
• Study First Posted: 2024-10-17
• Last Update Submitted: 2024-10-30
• Last Update Posted: 2024-11-01

Recruitment & Eligibility

• Status: COMPLETED
• Sex: All
• Target Recruitment: 172

Inclusion Criteria

Not provided

Exclusion Criteria

• Non-compliance with the investigation program
• Failure to sign the informed consent form
• Liver biopsy/clinic discrepancy
• For LMN a TMA "stress test": fish, see fruit and citrus fruit allergy
• Pregnancy
• For bile acid analysis: Treatment with BA or BA sequestrants
• For bile acid analysis: Portal hypertension (does not apply for MASH patients)
• For bile acid analysis: Cirrhosis (does not apply for MASH patients)
• For plasma spectroscopy: Cirrhosis
• Anamnesis of alcohol abuse (based on GGT, carbohydrate-deficient transferrin, urinary ethyl-glucuronide, and patient's history)

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Primary Outcome Measures

Name	Time	Method
Serum bile acids concentrations in the MASH/ Steatosis/ Healthy controls	A single day during within the data collection period = between September 2021 and September 2024	Individual bile acid concentrations in blood serum accessed by LC-MS/MS, measured in µmol/L or expressed as detectable or undetectable
The effect of serum bile acids in the MASH/ Steatosis/ Healthy controls discrimination	A single day during within the data collection period = between September 2021 to September 2024	To access the effect of serum bile acid concentrations on intergroup discrimination in a discriminant model as measured by model parameters and ROC
Native breath concentrations of volatile organic compounds in the aim groups	Single measurement on one day within 6 weeks (from mid-February 2023 to the end of March 2023)	Concentrations of VOC in fasting exhaled breath expressed in ppbv
A change in d-Limonene and TMA concentrations before and after the ingestion of capsules containing d-LMN, TMA, ppbv	Baseline, at 150min after capsules ingestion - both on one day within 6 weeks (from mid-February 2023 to the end of March 2023)	A change in concentration of d-Limonene and TMA in exhaled breath after an overnight fast/ post-ingestion of d-Limonene and TMA-containing capsules, expressed in ppbv
The effect of concentrations of volatile organic compounds in exhaled breath, native and post-ingestion, on the discrimination of the aim groups	On one day within 6 weeks (from mid-February 2023 to the end of March 2023)	To access the effect of VOC concentrations in exhaled breath, native and after the ingestion of d-Limonene and TMA-containing capsules, on intergroup discrimination in a discriminant model as measured by model parameters and ROC
Peak of d-LMN, and TMA concentration after the ingestion of capsules in the aim groups	At an individually specific time point for each participant that occurs within 4 hours after ingestion of d-Limonene and TMA-containing capsules on a single day within 6 weeks (from mid-February 2023 to the end of March 2023)	A maximum concentration of d-Limonene and TMA in exhaled breath post-ingestion of d-Limonene and TMA-containing capsules, expressed in ppbv
Spectroscopic patterns of blood plasma	Based on single peripheral blood uptake within the data collection period = between September 2021 and September 2024	Disease-specific patterns of blood plasma expressed as differences in spectra (their normalized intensity (a.u.)) and regions between the aim groups

Trial Locations

Locations (2)

Regional Hospital Liberec; 🇨🇿 Liberec, Czech Republic

General University Hospital in Prague; 🇨🇿 Praha 2, Czech Republic