HealthyNCT06647095
New Non-invasive Biomarkers of Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD)
An observational study of Healthy and Simple Steatosis (SS), run by General University Hospital, Prague at 2 sites in Czechia.
Trial 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. Read more about this study
- Who is it for?
- Ages 18 to 80 yearsHealthy volunteers can take part
- Conditions the study is for
- Healthy, Simple Steatosis (SS), Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD)
- What is being tested?
- D-Limonene Gelcaps and Fish Oil Concentrate, 1000 Mg Oral Capsule
- Type of treatment
- Pill or oral medicine
- How many people
- 172 participants
- Where
- 2 sitesCzechia
- Study dates
- 1 Sep 2021to 1 Sep 2024
- Phase
- Observational
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 80 years
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
Study population: Patients with MASLD of primary care clinic, healthy volunteers
Who may be able to join
- Presence of liver steatosis (detected either by hepatic ultrasonography/CAP/histologically) or MASH/simple steatosis confirmed by liver biopsy
- Absence of secondary causes of fat accumulation in the liver and other liver disease: ruled out other etiologies of liver disease, such as viral hepatitis, drug-induced liver disease, autoimmune liver disease, biliary tract disease, and hereditary metabolic diseases
- Absence of liver disease
- For breath analysis: absence of liver steatosis (normal liver ultrasound image and CAP less than 248 dB/m)
- For plasma spectroscopy and bile acid analysis: Body Mass Index ≤ 25 and waist-hip ratio ≤ 0.95 plus normal liver ultrasound
- Absence of diabetes mellitus and metabolic syndrome
- Normal liver function tests, lipid spectrum, fasting glycemia
- Alcohol intake less than 20 g/ day (women) or 30 g/ day (men)
Who cannot join
- 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
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- 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)
Trial locations and contacts
2 sites in 1 country.
Czechia2 sites
Regional Hospital Liberec
Liberec, Czech Republic, 46001
General University Hospital in Prague
Prague, Czech Republic, 12800
About this study
Official title: Identification of New Non-invasive Biomarkers of Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD) in Patients by Blood Plasma Spectroscopy, Breath Volatile Organic Compounds Analysis and Serum Bile Acids Analysis
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:
- Can a method find participants with higher liver fat than healthy participants?
- Can a method find participants in whom higher liver fat was a cause of liver inflammation or stiffness?
Read more
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)
Read the 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:
- analyzing the fasting spectrum of serum bile acids using liquid chromatography-mass spectrometry
- analyzing the disease-specific spectroscopy patterns given by vibrational and chiroptical spectroscopy of blood plasma
- 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.
- Conditions
- Healthy, Simple Steatosis (SS), Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD), Metabolic Dysfunction-associated Steatohepatitis (MASH)
- Being tested
- D-Limonene Gelcaps, Fish Oil Concentrate, 1000 Mg Oral Capsule
- Keywords
- Liver steatosis, Liver fibrosis, MASLD, MASH, Blood spectroscopy, Breath tests, d-Limonene, SYFT-MS, Bile acids, metabolic dysfunction-associated steatotic liver disease, metabolic dysfunction-associated steatohepatitis
- Medical subject headings
- Fatty Liver, Liver Cirrhosis
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
D-Limonene GelcapsDietary supplement
Capsules containing d-Limonene will be optionally given during the breath test to all participants of all groups
Fish Oil Concentrate, 1000 Mg Oral CapsuleDietary supplement
Capsules containing fish oil will be optionally given during the breath test to all participants of all groups
| Study group | Role | What they receive |
|---|---|---|
| Healthy controls | Not described Receives: Dietary Supplement: D-Limonene Gelcaps; Dietary Supplement: Fish Oil Concentrate, 1000 Mg Oral Capsule | |
| MASH | Not described Receives: Dietary Supplement: D-Limonene Gelcaps; Dietary Supplement: Fish Oil Concentrate, 1000 Mg Oral Capsule | |
| Steatosis | Not described Receives: Dietary Supplement: D-Limonene Gelcaps; Dietary Supplement: Fish Oil Concentrate, 1000 Mg Oral Capsule |
What is being measured
The primary outcome is the main question the study is designed to answer.
Serum bile acids concentrations in the MASH/ Steatosis/ Healthy controls
Individual bile acid concentrations in blood serum accessed by LC-MS/MS, measured in µmol/L or expressed as detectable or undetectable
Measured: A single day during within the data collection period = between September 2021 and September 2024
The effect of serum bile acids in the MASH/ Steatosis/ Healthy controls discrimination
To access the effect of serum bile acid concentrations on intergroup discrimination in a discriminant model as measured by model parameters and ROC
Measured: A single day during within the data collection period = between September 2021 to September 2024
Native breath concentrations of volatile organic compounds in the aim groups
Concentrations of VOC in fasting exhaled breath expressed in ppbv
Measured: Single measurement on one day within 6 weeks (from mid-February 2023 to the end of March 2023)
A change in d-Limonene and TMA concentrations before and after the ingestion of capsules containing d-LMN, TMA, ppbv
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
Measured: Baseline, at 150min after capsules ingestion - both on one day within 6 weeks (from mid-February 2023 to the end of March 2023)
The effect of concentrations of volatile organic compounds in exhaled breath, native and post-ingestion, on the discrimination of the aim groups
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
Measured: On one day within 6 weeks (from mid-February 2023 to the end of March 2023)
Peak of d-LMN, and TMA concentration after the ingestion of capsules in the aim groups
A maximum concentration of d-Limonene and TMA in exhaled breath post-ingestion of d-Limonene and TMA-containing capsules, expressed in ppbv
Measured: 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)
Spectroscopic patterns of blood plasma
Disease-specific patterns of blood plasma expressed as differences in spectra (their normalized intensity (a.u.)) and regions between the aim groups
Measured: Based on single peripheral blood uptake within the data collection period = between September 2021 and September 2024
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted17 Oct 2024
- Study start1 Sep 2021
- Primary completion1 Sep 2022
- Study completion1 Sep 2024
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (7)Papers the sponsor has linked to this study.
- Other trials for HealthyStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Retrospective
- Enrolment
- 172 participants (actual)
Sponsor and investigators
- Lead sponsor
- General University Hospital, Prague
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Barbora Nováková, MD, General University Hospital, Prague
- Investigators
- Barbora Nováková, MD, Principal investigator, General University Hospital, Prague
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT06647095
- Sponsor's protocol ID
- GUHPrague83/21
- Status last verified
- Oct 2024
- First submitted
- 15 Oct 2024
- Last updated
- 1 Nov 2024
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Undecided
Publications
Puri P, Daita K, Joyce A, Mirshahi F, Santhekadur PK, Cazanave S, Luketic VA, Siddiqui MS, Boyett S, Min HK, Kumar DP, Kohli R, Zhou H, Hylemon PB, Contos MJ, Idowu M, Sanyal AJ. The presence and severity of nonalcoholic steatohepatitis is associated with specific changes in circulating bile acids. Hepatology. 2018 Feb;67(2):534-548. doi: 10.1002/hep.29359. Epub 2017 Dec 23.
PubMed 28696585Spanel P, Smith D. Quantification of volatile metabolites in exhaled breath by selected ion flow tube mass spectrometry, SIFT-MS. Clin Mass Spectrom. 2020 Feb 13;16:18-24. doi: 10.1016/j.clinms.2020.02.001. eCollection 2020 Apr.
PubMed 34820516Sinha R, Lockman KA, Homer NZM, Bower E, Brinkman P, Knobel HH, Fallowfield JA, Jaap AJ, Hayes PC, Plevris JN. Volatomic analysis identifies compounds that can stratify non-alcoholic fatty liver disease. JHEP Rep. 2020 Jun 15;2(5):100137. doi: 10.1016/j.jhepr.2020.100137. eCollection 2020 Oct.
PubMed 32775974Ten-Doménech I, Rienda I, Pérez-Rojas J, et al. Progress and challenges of mid-infrared spectroscopy for liver characterization focusing on steatosis, fibrosis and cancer. Applied Spectroscopy Reviews. 2024;59(4):578-599
Targher G, Byrne CD, Tilg H. MASLD: a systemic metabolic disorder with cardiovascular and malignant complications. Gut. 2024 Mar 7;73(4):691-702. doi: 10.1136/gutjnl-2023-330595.
PubMed 38228377Zizalova K, Novakova B, Vecka M, Petrtyl J, Lanska V, Pelinkova K, Smid V, Bruha R, Vitek L, Lenicek M. Serum concentration of taurochenodeoxycholic acid predicts clinically significant portal hypertension. Liver Int. 2023 Apr;43(4):888-895. doi: 10.1111/liv.15481. Epub 2022 Nov 25.
PubMed 36433660WED-280 Spectroscopy of blood plasma has the potential to differentiate metabolic dysfunction-associated steatohepatitis from steatosis Nováková, Barbora et al. Journal of Hepatology, Volume 80, S539 - S540