Probiotics and Oxytocin Nasal Spray on Social Behaviors of Autism Spectrum Disorder (ASD) Children

Study Overview
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Brief Summary
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Because oral probiotics reported to potentially induce endogenous Oxytocin, and Oxytocin has been reported to improve social behaviors, the investigators will conduct a pilot trial to compare the effects of probiotics and Oxytocin on social behavioral changes in ASD children. Additionally, the investigators will check oxytocin levels, and perform brain fMRI in some subjects, in order to determine which treatment is more efficient, sustainable, and practical, and whether both treatments in combination are better than either treatment alone. If the trial is conclusive, the investigators will conduct a trial in large scale to understand more the mechanism of ASD behaviors and corresponding effective interventions.

Detailed Description
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This study description is in accordance with the Consolidated Standards of Reporting Trials (CONSORT) guidelines that are published for the evaluation of randomized controlled trials. This clinical trial is a randomized, double-blind placebo controlled study. Subjects will be randomized to 2 groups: Phase 1: a. oral placebo, and b. oral probiotics; Phase 2: a. intranasal Oxytocin(OXT) + oral placebo, and b. intranasal OXT + oral probiotics The treatment will proceed for a total of 28 weeks. In the first phase (16 weeks), all the patients will be randomly and proportionally divided into two groups: Group A (30 subjects) receives oral probiotics while Group B (30 subjects) receives an oral placebo. In the second phase, subjects in Group A and Group B will continue their respective oral probiotics or placebo administration as in Phase 1. In addition, both groups will be simultaneously administered with intranasal OXT spray. Testing will be performed 3 times total (before, during, and after treatment). The tests include behavioral surveys, cognitive tests, clinical autonomic tests, and blood tests for oxytocin levels. Investigators plan to select up to 10 subjects from each group to conduct a series of MRI studies at week 0, week 16 and week 28.

Official Title
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The Effects of Probiotics and Oxytocin Nasal Spray on Social Behaviors of ASD Children- A Pilot Study

Conditions
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Autism Spectrum Disorder

Intervention / Treatment
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* Drug: intranasal oxytocin
* Dietary Supplement: oral probiotics
* Dietary Supplement: oral placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Age between 3-25years old; Pre-existing diagnosis of autism; subjects may be asked to provide documentation confirming diagnosis by DSM-V-TR criteria, ADOS, ADI-R, or other clinical forms A care provider who can reliably bring the participant to study visits; No planned changes in medications or psychosocial interventions during trial (stable medications within the last 2 weeks); Willingness to provide blood samples Exclusion Criteria: Pregnant woman (before or during the study). Comorbidity of other neurological and/or psychiatric disorders such as unstable seizures, schizophrenia, schizoaffective disorder, bipolar disorders or history of substance abuse. Psychotropic medication use Subjects with active cardiovascular disease that is not controlled by medication. Oxytocin, antibiotic, or probiotic use within the last 30 days. Regular nasal obstruction or nosebleeds Significant hearing, vision, or motor impairments Habitual consumption of large volumes of water Started taking new medications within the last 2 weeks

Ages Eligible for Study
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Minimum Age: 3 Years
Maximum Age: 25 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Phase 1: a. oral placebo, and b. oral probiotics; Phase 2: a. intranasal OXT + oral placebo, and b. intranasal OXT + oral probiotics
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: oral probiotics and oxytocin spray<br>Subjects will receive oral probiotics, 2 pills per day, for 28 weeks. For the last 12 weeks, subjects will also receive intranasal oxytocin spray at the following dose: 4 IU in week 1, 8 IU in week 2, 16 IU in week 3, and 24 IU in weeks 4-12. | Drug: intranasal oxytocin<br>* 4-24 IU per day, dosage gradually increases<br>* Other names: Novartis Syntocinon;Dietary Supplement: oral probiotics<br>* 200 million cfu per day<br>|
| Placebo Comparator: oral placebo and oxytocin spray<br>Subjects will receive oral placebo, 2 pills per day, for 28 weeks. For the last 12 weeks, subjects will also receive intranasal oxytocin spray at the following dose: 4 IU in week 1, 8 IU in week 2, 16 IU in week 3, and 24 IU in weeks 4-12. | Drug: intranasal oxytocin<br>* 4-24 IU per day, dosage gradually increases<br>* Other names: Novartis Syntocinon;Dietary Supplement: oral placebo<br>* 2 pills per day<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Social Responsiveness Scale (SRS) Edition 2 | social communication and behavior - The scoring is based on T-score which is based on the sum of responses as follows (76 to higher - severe, 66 to 75- moderate deficiencies, 60 to 65 - mild deficiencies, 59 and below is not clinically significant for ASD). | change from baseline at 0, 16, and 28 weeks | 
| Aberrant Behavior Checklist (ABC) Edition 2 | social behavior test - The total score is calculated based on 5 sub-scales (Irritability, Social Withdrawal, Stereotypic Behavior, Hyperactive/Noncompliance, and Inappropriate Speech). There are 58 questions that are scored on a 0-3 scale 0 -not at all a problem, 1- the behavior is a problem but slight in degree, 2- the problem is moderately serious and 3- the problem is severe in degree. Based on this sub-scores are calculated and added to get the total score. | change from baseline at 0, 16, and 28 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Neuroinflammation and Oxytocin levels | neuroendocrine biomarker measured in blood ( Melatonin, Oxytocin, Tumour Necrosis Factor alpha, testosterone and Interleukin 6) | change from baseline at 0, 16, and 28 weeks | 
| structural MRI | The software Freesurfer will be used to calculate volume of brain regions and diffusion parameters (e.g. fractional anisotropy and mean diffusivity) from structural T1 and diffusion tensor images respectively. T-tests will be applied for group comparisons. | change from baseline at 0, 16, and 28 weeks | 
| Functional MRI (resting state) | Correlation analysis will be used to calculate the connectivity across different brain regions. T-tests will be used for group comparisons. | change from baseline at 0, 16, and 28 weeks | 
| Functional MRI (task based) | General linear modeling will be used to calculate brain responses to the tasks. T-tests will be used for group comparisons. | change from baseline at 0, 16, and 28 weeks | 
| Autonomic indices 1 | Blood volume pulse | change from baseline at 0, 16, and 28 weeks | 
| Autonomic indices 2 | heart rate | change from baseline at 0, 16, and 28 weeks | 
| Autonomic indices 3 | peripheral skin temperature | change from baseline at 0, 16, and 28 weeks | 
| Autonomic indices 4 | skin electrodermal activity | change from baseline at 0, 16, and 28 weeks | 
| Autonomic indices 5 | blood oxygen saturation | change from baseline at 0, 16, and 28 weeks | 
| Microbiome | 16s metagenomic sequencing of the microbiome | change from baseline at 0, 16, and 28 weeks | 
| Eye tracking and Behavioral task (joint attention) | joint attention - Conduct T-tests and calculate P values for total and average fixation values for each area of interest | change from baseline at 0, 16, and 28 weeks | 
| Eye tracking and behavioral task (emotion response) | emotion response - The accuracy and reaction time will be calculated for each time point. Then an ANOVA will be used to compare the results. | change from baseline at 0, 16, and 28 weeks | 
| Eye tracking and behavioral task (eye behavior) | eye behavior - Conduct T-tests and calculate P values for total and average fixation values for each area of interest | change from baseline at 0, 16, and 28 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Probiotics, Oxytocin