Butyrate Supplementation in Children With Autism Spectrum Disorder (ASD) and Functional Gastrointestinal Disorders
NCT Number: NCT06514573
Acronym: b(AUT)yrate
Status: NOT_YET_RECRUITING
Study Type: INTERVENTIONAL
Phases: NA
Conditions: Autism Spectrum Disorder (ASD)
Interventions: DIETARY_SUPPLEMENT: Sodium Butyrate|DIETARY_SUPPLEMENT: Placebo (cornstarch)
Sponsor: Istituto Superiore di Sanità
Collaborators: University of Rome Tor Vergata|Federico II University
Sex: ALL
Age: CHILD
Enrollment: 128
Study Design: Allocation: RANDOMIZED|Intervention Model: PARALLEL|Masking: QUADRUPLE (PARTICIPANT, CARE_PROVIDER, INVESTIGATOR, OUTCOMES_ASSESSOR)|Primary Purpose: TREATMENT
Start Date: 2024-09-02
Completion Date: 2025-07-28
Locations: Azienda Ospedaliera Universitaria Federico II, Naples, 80131, Italy|Policlinico Tor Vergata Hospital, Rome, 00133, Italy|Istituto Superiore di Sanità, Rome, 00161, Italy

Summary:
Autism spectrum disorder (ASD) is a neurodevelopmental condition occurring in 1:77 Italian children. Several comorbidities are reported, including functional gastrointestinal disorders (FGIDs) present in up to 70% of patients. FGIDs are disorders resulting from a combination of symptoms affecting motility, hypersensitivity, and other functions, which are not caused by anatomic or organic origin and that impact the severity of ASD core symptoms and complicate the clinical management of ASD children, especially those who are non-verbal. Evidence reports gut microbiome (GM) remodelling in ASD children, and postbiotic butyrate, a GM-derived metabolite, attenuates FGIDs in children and restores social behavior in ASD mouse models. Clinical data on butyrate effects in ASD are still scanty. The present study investigates the therapeutic effects of a 16-week oral postbiotic supplementation on clinical/behavioral profiles, gastrointestinal disturbances, gut microbiome, and immune and inflammatory biomarkers in peripheral blood and fecal samples in children with ASD and FGIDs. Using the Machine Learning (ML) approach, a subset of artificial intelligence, this study also aims to identify predictive factors implicated in the effect of the postbiotic supplementation on FGIDs, important for prevention through modulation of the microbiota. The investigators expect that treating FGIDs will have an impact on the behavioral and core symptoms of ASD and the quality of life of children and their families.

Primary Outcome Measures: Gastrointestinal symptom severity., Rate of subjects presenting a clinically relevant improvement in Functional Gastrointestinal Disorders (FGIDs) severity, measured by 6-item Gastrointestinal Severity Index (6-GSI) score decrease \>4 points.

The Gastrointestinal Severity Index (6-GSI) score is the sum of the individual item scores.

The higher it is, the worse is the outcome. Score range for each item: 0, 1, 2 (0 no symptoms, 1 mild symptoms, 2 severe symptoms)., At baseline (Timepoint 0) and at 4 months (Timepoint 1)
Secondary Outcome Measures: Incidence of Treatment-Emergent Adverse Events [Safety and Tolerability], Parental questionnaires on the presence of nausea, skin irritation, itching, patches on the skin, stomach ache, flatulence, meteorism, vomiting, constipation, increased/decreased appetite, heartburn, reflux, burping.

Measured in hours or days of symptoms' duration., At 4 months (Timepoint 1) and at 8 months (Timepoint 2)|Unexplained daytime irritability, Parental questionnaires on the presence of angry outbursts, bouts of crying, easy frustration, mood swings, aggression, restlessness.

Measured by symptom intensity during daytime: light, mild, severe., At 4 months (Timepoint 1) and at 8 months (Timepoint 2)|Nocturnal awakening, Parental questionnaires on the presence of night waking. Measured as Yes (number of episodes) or No., At 4 months (Timepoint 1) and at 8 months (Timepoint 2)|Persistency of Functional Gastrointestinal Disorders severity improvement, Measured by 6-item Gastrointestinal Severity Index (6-GSI) score: the sum of the individual item scores.

The higher it is, the worse is the outcome. Score range for each item: 0, 1, 2 (0 no symptoms, 1 mild symptoms, 2 severe symptoms)., At 4 months (Timepoint 1) and at 8 months (Timepoint 2)|Gut Microbiota structure and function (fecal Short-Chain Fatty Acids levels, SCFAs), Bacterial DNA extraction from one fecal sample will be carried out by shotgun metagenomics. Fecal SCFAs (acetate, propionate and butyrate) levels will be measured using the second fecal sample by gas chromatograph-mass spectrometry and expressed in mM (millimolar) with a range of 0-∞., At baseline (Timepoint 0) and at 4 months (Timepoint 1)|Changes in Autism Spectrum Disorder severity, Measured by Social Responsiveness Scale-2 (SRS-2). Score range for each item: 0-100. The higher the score, the worse the outcome., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Changes in Autism Spectrum Disorder behavior, Measured by Aberrant Behavior Checklist (ABC). Score range for each item: 0-999. The higher the score, the worse the outcome., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Changes in Autism Spectrum Disorder symptomatology: repetitive behaviors, Delta of scores at Repetitive Behavior Scale-Revised (RBS-R), a 43-item scale. Score range for each item: 0-100. Higher scores are related to greater severity of symptoms., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Changes in Sensory input reaction, Measured by Sensory Profile 2 (SP-2) scores. Score range for raw score: 86-430. Score range for percentiles: 0-100. The further away from the mean score, the more pathological.

Score categories for sensitivity level: 1,2,3,4,5.

1. Severe hyposensitivity (\< -2 Standard Deviation)
2. Mild hyposensitivity (between -1 e -2 Standard Deviation)
3. Typical sensitivity (between -1 and 1 Standard Deviation)
4. Mild hypersensitivity (between 1 and 2 Standard Deviation)
5. Severe hypersensitivity (\> 2 Standard Deviation), At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Changes in Adaptive Functioning, Measured by Vineland Adaptive Behavior Scales-II (VABS-II). Score range for each item: 0-160. The higher the score, the better the outcome., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Changes in emotional/behavioral problems, Measured by Child Behavior Checklist (CBCL syndrome subscales). Score range for each item: 0-999. The higher the score, the worse the outcome., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Changes in sleep disorders, Measured by Children's Sleep Habits Questionnaire Abbreviated (CSHQ).

Score ranges for items:

Bedtime (0-63), Sleep behavior (0-49), Waking during the night (0-14), Morning wake up (0-28).

Total sleep disturbance score: the sum of the individual item scores (0-154). The higher the score, the worse the outcome., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Sleep disturbance assessment, Measured by Sleep Disturbance Scale for Children (SDSC).

Score ranges for items:

Sleep onset and maintenance disorders (7-35), Sleep breathing disorders (3-15), Arousal disorders (3-15), Wake-Sleep Transition Disorder (6-30), Excessive sleepiness disorder (5-25), Nocturnal hyperhidrosis (2-10).

Total score: the sum of the individual item scores (26-130). Higher scores indicate more acute sleep disturbances., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Impact on parental quality of life, Evaluated through the Quality of life questionnaire (EUROHIS QOL-8). Score range for each item: 1-5. Higher score indicates a better condition. Overall index score: the mean of the individual scores, ranging from 1 to 5 with a higher score indicating a better quality of life., At baseline (Timepoint 0), at 4 months (Timepoint 1) and 8 months (Timepoint 2)|Serum proinflammatory and regulatory cytokines, Intracellular expression of cytokines (IL-1beta, IL-5, IL-6, IL-10, IL-15, IL-17, TGF-beta, TNF-alpha, IFN-alpha, IFN-gamma) measured by Enzyme-Linked Immuno Assay (ELISA).

All reported as pg/ml., At baseline (Timepoint 0) and at 4 months (Timepoint 1)|Plasmatic levels of chemochines involved in the migration of inflammatory cells within the CNS (Part 1), Expression of ICAM-1, PECAM-1, and P-selectin measured by Enzyme-Linked Immuno Assay (ELISA).

All reported as ng/ml., At baseline (Timepoint 0) and at 4 months (Timepoint 1)|Plasmatic levels of chemochines involved in the migration of inflammatory cells within the CNS (Part 2), Expression of L-selectin measured by Enzyme-Linked Immuno Assay (ELISA). Reported as pg/ml., At baseline (Timepoint 0) and at 4 months (Timepoint 1)|Peripheral blood mononuclear cells features (Part 1), Characterization of peripheral blood mononuclear cells immunophenotype by flow cytometry (Cytoflex, Beckman Coulter).

All expressed in % (range 0,000-100,000):

Lymphocytes, Monocytes, Granulocytes,CD3, CD4, CD8, CD19, CD16, CD56, HLA-DR Monocytes, HLA-DR Lymphocytes, HLADRMONOLYMPH, CD169Leukocytes, CD169Lymphocytes, CD169Monocytes, CD169Granulocytes, HLADRLLeukocytes, HLADRLymphocytes, HLADRMonocytes, HLADRGranulocytes, HLADRCD169Leukocytes, HLADRCD169Lymphocytes, HLADRCD169Monocytes, HLADRCD169Granulocytes, CD3_tube2, CD4_tube2, CD4CM, CD4NAIVE, CD4EM, CD4TEM, CD8_tube2, CD8CM, CD8NAIVE, CD8EM, CD8TEM, CD4PD1CD57, CD4CD57, CD4PD1, CD8PD1CD57, CD8CD57, CD8PD1, CD19_tube2, Marginal, NaiveB, IgDmCD27m, IgDmCD27p, CD21lowCD38low, IgMpIgDp, IgMmIgDp, IgDmIgMm, IgMpIgDm, IgMpCd27pCD38p, Unswitchedmemory, IgMpCD27mCD38dim, IgMpCD27mCD38high, IgDmIgMmCD27pCD38high, Switchedmemory, IgMmIgDmCD27mCD38low, IgMmIgDmCD27mCD38high, Plasmablasts, IgMpCD27m, TransitionalBcells, At baseline (Timepoint 0) and at 4 months (Timepoint 1)|Peripheral blood mononuclear cells features (Part 2), Characterization of peripheral blood mononuclear cells immunophenotype by flow cytometry (Cytoflex, Beckman Coulter).

All expressed as absolute number/ml:

CD169MEDLYMPH, CD169MEDMONO, CD64MEDLYMPH, CD64MEDNEUTRO, At baseline (Timepoint 0) and at 4 months (Timepoint 1)|Peripheral blood mononuclear cells features (Part 3), Characterization of peripheral blood mononuclear cells immunophenotype by flow cytometry (Cytoflex, Beckman Coulter).

All expressed as Ratio, absolute number:

CD169 Monocytes/Lymphocytes RMFI, CD64 Neutrophils/Lymphocytes RMFI, At baseline (Timepoint 0) and at 4 months (Timepoint 1)