Use of a Selected Mixture of Probiotic Strains for Degrading Gluten During Digestion

March 22, 2023 updated by: Free University of Bozen-Bolzano

Novel Probiotic Preparation With Gluten Degrading Activity and Gut Microbiota Modulating Effect

Gluten intake spreads worldwide, being the major food protein consumed in the Western diets (up to 20 g gluten/d). But gluten has unique and unusual features. It resists the complete luminal digestion by gastric, pancreatic and intestinal brush border enzymes, and is susceptible to post-translational modification (deamidation) by mucosal transglutaminases. Apart from partial digestion, gluten per se has a negative impact on a consistent part of the worldwide population, which mainly results in the manifestations of celiac disease (CD) or other gluten-related disorders. This study will enable to test in vivo a novel multi-species probiotic that in vitro has proven to degrade gluten to non-immunotoxic peptides.

Study Overview

Status

Status

Indicates the current recruitment status or the expanded access status.
Completed

Conditions

Conditions

The disease, disorder, syndrome, illness, or injury that is being studied. On ClinicalTrials.gov, conditions may also include other health-related issues, such as lifespan, quality of life, and health risks.

Intervention / Treatment

Intervention / Treatment

A process or action that is the focus of a clinical study. Interventions include drugs, medical devices, procedures, vaccines, and other products that are either investigational or already available. Interventions can also include noninvasive approaches, such as education or modifying diet and exercise.

Detailed Description

Hypothesis and Significance:

The project intends to confirm whether the novel multi-species probiotic preparation has the ability to degrade gluten upon digestion at increasing dosages (from 50mg/d up to 10g/d). Further we aim to evaluate the efficiency of the probiotic to persist and colonize the human gut as well as its ability to modulate the human gut microbiome.

Since this is a phase 1 trial, healthy participants will be recruited to avoid any triggers of CD symptoms. Participants will undergo gluten free diet for 10 days to eliminate any traces of gluten from their feces and will be provided probiotic/placebo capsules as long as specific amounts of gluten to be ingested with their meal. Faecal samples will be collected at the end of each period that increasing gluten amounts were ingested. Residual gluten amounts in feces will be evaluated and the fecal microbiome will be studied by 16S metabarcoding analysis. Fecal metabolome will be also assessed as long as the persistence and colonisation ability of the probiotic preparation by qPCR.

Specific Aim 1:

Evaluate the gluten depredating efficiency of the probiotic by ELISA

Specific Aim 2:

Investigate the gut microbiome alterations between the intervention groups and possible modulation

Specific Aim 2:

Investigate the fecal metabolome (Volatiles and short chain fatty acids) between intervention groups

Specific Aim 2:

Monitor the persistence and colonization ability of the probiotic preparation by qPCR

Study Type

Study Type

Describes the nature of a clinical study. Study types include interventional studies (also called clinical trials), observational studies (including patient registries), and expanded access.
Interventional

Enrollment (Actual)

Enrollment

The number of participants in a clinical study. The "anticipated" enrollment is the target number of participants that the researchers need for the study.
70

Phase

Phase

The stage of a clinical trial studying a drug or biological product, based on definitions developed by the U.S. Food and Drug Administration (FDA). The phase is based on the study's objective, the number of participants, and other characteristics. There are five phases: Early Phase 1 (formerly listed as Phase 0), Phase 1, Phase 2, Phase 3, and Phase 4. Not Applicable is used to describe trials without FDA-defined phases, including trials of devices or behavioral interventions.
  • Not Applicable

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Locations

  • Italy
      • Bolzano, Italy, 39100
        • Free University of Bolzano-Bozen

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Eligibility Criteria

The key requirements that people who want to participate in a clinical study must meet or the characteristics they must have. Eligibility criteria consist of both inclusion criteria (which are required for a person to participate in the study) and exclusion criteria (which prevent a person from participating). Types of eligibility criteria include whether a study accepts healthy volunteers, has age or age group requirements, or is limited by sex.

Ages Eligible for Study

18 years to 65 years (Adult, Older Adult)

Accepts Healthy Volunteers

Yes

Genders Eligible for Study

All

Description

Inclusion Criteria:

  • healthy individuals;
  • adherent to Mediterranean diet

Exclusion Criteria:

  • known medical disease;
  • known digestive disease symptoms;
  • known family history of celiac disease (CD);
  • wheat allergy;
  • and use of prescription medications (including antibiotics or probiotics in the previous 2 months)

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

  • Primary Purpose: Screening
  • Allocation: Randomized
  • Interventional Model: Parallel Assignment
  • Masking: Triple

Arms and Interventions

Participant Group / Arm

Participant Group / Arm

A group or subgroup of participants in a clinical trial that receives a specific intervention/treatment, or no intervention, according to the trial's protocol.
Intervention / Treatment

Intervention / Treatment

A process or action that is the focus of a clinical study. Interventions include drugs, medical devices, procedures, vaccines, and other products that are either investigational or already available. Interventions can also include noninvasive approaches, such as education or modifying diet and exercise.
Active Comparator: PR
Fifty healthy volunteers were randomly allocated in the probiotic arm. To eliminate residual traces of gluten and similar proteins from the faecal material, both groups underwent a gluten-free diet (GFD) from day-1 to day-10. After 10 days, gluten administration started. The increasing administration plan was as follows: 50 mg/day for 4 days; 1 g/day for subsequent 4 days; 3 g/day for subsequent 4 days; and 10 g/day (in this case, reintroducing an equivalent amount of wheat-based bread - 4 slices) for subsequent 20 days. At this stage (10 + 4 + 4 + 4 + 10 days = total of 32 days), the administration of the probiotic preparation was interrupted, with a period of 10 days of wash-out.
Combination product: Probiotic administration and gluten
Probiotic preparation including multi-species strains of Lactobacillus and Bacillus. The probiotic was administered at baseline and interrupted after 32 days. At the same time the other arm received placebo. Gluten was provided after 10 days of gluten free diet in increasing amounts
Placebo Comparator: PL
Twenty healthy volunteers were randomly allocated in the placebo.To eliminate residual traces of gluten and similar proteins from the faecal material, both groups underwent a gluten-free diet (GFD) from day-1 to day-10. After 10 days, gluten administration started. The increasing administration plan was as follows: 50 mg/day for 4 days; 1 g/day for subsequent 4 days; 3 g/day for subsequent 4 days; and 10 g/day (in this case, reintroducing an equivalent amount of wheat-based bread - 4 slices) for subsequent 20 days. At this stage (10 + 4 + 4 + 4 + 10 days = total of 32 days), the administration of placebo preparation was interrupted, with a period of 10 days of wash-out.
Combination product: Placebo administration and gluten
The placebo was administered at baseline and interrupted after 32 days. At the same time the other arm received probiotic. Gluten was provided after 10 days of gluten free diet in increasing amounts

What is the study measuring?

Primary Outcome Measures

Primary Outcome Measures

In a clinical study's protocol, the planned outcome measure that is the most important for evaluating the effect of an intervention/treatment. Most clinical studies have one primary outcome measure, but some have more than one.
Outcome Measure
Measure Description
Time Frame
Hydrolyzed gluten amount in feces by competitive Elisa R5 antibody (gluten ppm)
Time Frame: 6 months
The collection of faecal samples will be at baseline (T0); 10 days of GFD (T1); 4 days of 50 mg/day gluten intake (T2); 4 days of 1 g/day gluten intake (T3); 4 days of 3 g/day gluten intake (T4); 20 days of 10 g/day gluten intake (T5), of which 10 last days were the wash-out (T6).The gluten degrading efficiency of the probiotic will be assessed in all above mentioned time points with competitive Elisa using the R5 antibody for hydrolysed gluten (gliadin epitopes). The concentration of gliadin (ppm) will be then converted to gluten according to the multiplication factor (2) reported by the manufacturer.
6 months
Gut microbiome
Time Frame: 7 months
Fecal samples DNA from baseline, intervention period and wash out will be sequenced targeting the V4 region of 16s rRNA gene to evaluate the alterations of the gut microbiome and possible modulation in the PR arm due to the probiotic mixture administrated
7 months
Volatile compounds determination by GC-MS using SPME extraction
Time Frame: 6 months
Fecal samples at the beginning of intervention (T1), end of intervention (T5) and wash-out (T6) will be evaluated for their organic volatile compounds by GC-MS. Volatile compounds and short chain fatty acids composition will be compared between the intervention groups PL vs PR. 4-methyl-2pentanol (final concentration 1 mg L-1) will be used as an internal standard in all analyses, to quantify the identified compounds by interpolation of the relative areas versus internal standard area.
6 months
Short chain fatty acids determination by GC-MS using SPME extraction
Time Frame: 6 months
Fecal samples at the beginning of intervention (T1), end of intervention (T5) and wash-out (T6) will be evaluated for SCFA by GC-MS. SCFA compounds and short chain fatty acids composition will be compared between the intervention groups PL vs PR. A stock solution containing the mixture of SCFA standards (acetic acid, butyric acid, propionic acid, isobutyric acid and valeric acid) will be dissolved in ultrapure water to obtain a calibration curve ranging from 1 μg mL-1 to 250 μg mL-1.The calibration curve will be constructed by plotting the normalized peak area versus concentration of individual SCFA. The relative peak of SCFA in faecal sample will be integrated and the concentration of SCFA will be calculated by the calibration curve equation
6 months
Persistence and colonization ability of probiotic preparation by quantitative PCR (copy numbers)
Time Frame: 2 months
The persistence and colonization ability of the probiotic preparation will be evaluated by qPCR on cDNA and DNA at T1, T5 and T6. For each species belonging to the probiotic preparation, species- specific primers will be used. The qPCR results (cycle threshold, CT) will be converted in Copy Number (CN) based on standard curves previously constructed by using serial dilutions of DNA extracted from pure cultures. The CN and CN(Log) will be calculated based on DNA concentration and amplicon length. The standard curves will be obtained by CT and CN(Log) interpolation.
2 months

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Sponsor

The organization or person who initiates the study and who has authority and control over the study.
Free University of Bozen-Bolzano

Collaborators

Collaborators

An organization other than the sponsor that provides support for a clinical study. This support may include activities related to funding, design, implementation, data analysis, or reporting.
Evonik Operations GmbH, Germany

Investigators

Investigators

A researcher involved in a clinical study. Related terms include site principal investigator, site sub-investigator, study chair, study director, and study principal investigator.
  • Principal Investigator: Olga Nikoloudaki, Ph.D, Free University of Bolzano-Bozen

Publications and helpful links

The person responsible for entering information about the study voluntarily provides these publications. These may be about anything related to the study.

General Publications

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

Study Start

The actual date on which the first participant was enrolled in a clinical study. The "anticipated" study start date is the date that the researchers think will be the study start date.
October 1, 2020

Primary Completion (Actual)

Primary Completion

The date on which the last participant in a clinical study was examined or received an intervention to collect final data for the primary outcome measure. Whether the clinical study ended according to the protocol or was terminated does not affect this date. For clinical studies with more than one primary outcome measure with different completion dates, this term refers to the date on which data collection is completed for all the primary outcome measures. The "anticipated" primary completion date is the date that the researchers think will be the primary completion date for the study.
April 30, 2021

Study Completion (Actual)

Study Completion

The date on which the last participant in a clinical study was examined or received an intervention/treatment to collect final data for the primary outcome measures, secondary outcome measures, and adverse events (that is, the last participant's last visit). The "anticipated" study completion date is the date that the researchers think will be the study completion date.
May 31, 2021

Study Registration Dates

First Submitted

First Submitted

The date on which the study sponsor or investigator first submitted a study record to ClinicalTrials.gov. There is typically a delay of a few days between the first submitted date and the record's availability on ClinicalTrials.gov (the first posted date).
March 3, 2023

First Submitted That Met QC Criteria

First Submitted That Met QC Criteria

The date on which the study sponsor or investigator first submits a study record that is consistent with National Library of Medicine (NLM) quality control (QC) review criteria. The sponsor or investigator may need to revise and submit a study record one or more times before NLM's QC review criteria are met. It is the responsibility of the sponsor or investigator to ensure that the study record is consistent with the NLM QC review criteria.
March 22, 2023

First Posted (Actual)

First Posted

The date on which the study record was first available on ClinicalTrials.gov after National Library of Medicine (NLM) quality control (QC) review has concluded. There is typically a delay of a few days between the date the study sponsor or investigator submitted the study record and the first posted date.
April 4, 2023

Study Record Updates

Last Update Posted (Actual)

Last Update Posted

The most recent date on which changes to a study record were made available on ClinicalTrials.gov. There may be a delay between when the changes were submitted to ClinicalTrials.gov by the study's sponsor or investigator (the last update submitted date) and the last update posted date.
April 4, 2023

Last Update Submitted That Met QC Criteria

Last Update Submitted That Met QC Criteria

The most recent date on which the study sponsor or investigator submitted changes to a study record that are consistent with National Library of Medicine (NLM) quality control (QC) review criteria. It is the responsibility of the sponsor or investigator to ensure that the study record is consistent with the NLM QC review criteria.
March 22, 2023

Last Verified

Last Verified

The most recent date on which the study sponsor or investigator confirmed the information about a clinical study on ClinicalTrials.gov as accurate and current. If a study with a recruitment status of recruiting; not yet recruiting; or active, not recruiting has not been confirmed within the past 2 years, the study's recruitment status is shown as unknown.
March 1, 2023

More Information

Terms related to this study

Keywords

Other Study ID Numbers

Other Study ID Numbers

Identifiers or ID numbers other than the NCT number that are assigned to a clinical study by the study's sponsor, funders, or others. These numbers may include unique identifiers from other trial registries and National Institutes of Health grant numbers.
  • GlutProbiotic

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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