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OpenTrials
Completed

NCT Number: NCT05798689

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

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.

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Key information

Conditions

Probiotics Celiac Disease Digestive System Diseases Gastrointestinal Diseases Intestinal Diseases Malabsorption Syndromes Metabolic Diseases Nutritional and Metabolic Diseases

Age range

18 year–65 year

Sex eligibility

All sexes

Study type

Interventional

Phase

Not applicable

Primary location

Free University of Bolzano-Bozen

Bolzano, 39100, Italy

About this study

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

Who can participate

Healthy volunteers accepted: Yes

Only the study team can determine whether someone qualifies for participation.

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)

Treatment and study plan

Probiotic administration and gluten

Combination Product

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 administration and gluten

Combination Product

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

Primary outcomes

  1. 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.

  2. 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

  3. 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.

  4. 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

  5. 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.

Sponsors and collaborators

Lead sponsor

Free University of Bozen-Bolzano

Other

Collaborators

  • Evonik Operations GmbH, Germany

Registry information

Official study title

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

Important dates

Study start
2020
Primary completion
2021
Study completion
2021
First posted
Apr 4, 2023
Registry last updated
Apr 4, 2023

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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