Neural Biomarkers in Fragile X Syndrome

Completed

• Conditions: Neurotypical Adults; Fragile X Syndrome (FXS)

• Registration Number: NCT06957054
• Lead Sponsor: Connecta Therapeutics, S.L.

Brief Summary

The purpose of this non-interventional test /re-test study is to assess neural biomarkers in adult subjects with Fragile X syndrome compared to those measured in a population of typically developing adults

Detailed Description

This study consists of a screening period and two cross sectional evaluations assessed in one month interval (test/re-test), first on the Visit 1 (Basal) and the other on the Visit 2 (end-of-study visit, EOS).

Study Timeline

• Study Start: 2024-12-16
• Primary Completion: 2025-03-20
• Study Completion: 2025-03-20
• Status Verified: 2025-04
• Study First Submitted: 2025-04-10
• Study First Submitted QC: 2025-04-25
• Last Update Submitted: 2025-04-25
• Study First Posted: 2025-05-04
• Last Update Posted: 2025-05-04

Recruitment & Eligibility

• Status: COMPLETED
• Sex: All
• Target Recruitment: 20

Inclusion Criteria

• Clinical and molecular diagnosis of Fragile X syndrome (> 200 CGG repeats in the promoter region of the FMR1 gene).
• Legal representative understands and accepts the study procedures. If only one parent signs, he/she should confirm that the other parent does not object to the patient's participation in the research project.
• Subject assenting and/or willing to participate.
• Signed informed consent by a legal representative before any study-mandated procedure.
• Subject is independently mobile and has sufficient vision and hearing to participate in study evaluations. They must be able to be understood most of the time and must not use other forms of communication, signs, symbol boards or devices as their primary form of communication.
• Subjects must have a parent, or other reliable caregiver, who agrees to accompany the subject to all study visits, provide information about the subject as required by the protocol, and ensure compliance with study tests.
• Subjects are expected to complete all procedures scheduled during the study visits.

Subjects with Fragile X syndrome

Exclusion Criteria

• Personal history of infantile spasms/convulsions/epilepsy, severe head trauma or CNS infections (e.g. meningitis), with the exception of infantile febrile seizures.
• Subjects with a current diagnosis including severe autism spectrum disorder or any primary psychiatric diagnosis according to DSM-5 (Diagnostic and Statistical Manual of Mental Disorders-DSM-5). Diagnoses that are secondary, such as attention deficit hyperactivity disorder, depressive disorders and conduct disorders are allowed as long as they are considered to not interfere with study conduct and are stable during the 3 months before inclusion. Related allowed treatments must be on stable dosing for the last 3 months.
• Substance use disorder according to the DSM-5 criteria.
• Epileptiform abnormalities (excluding isolated sharp waves and beyond those expected for age).
• Any life-threatening disease.
• Any other clinically relevant concomitant disease or condition or finding at screening that in the judgment of the investigator could interfere with, the treatment thereof might interfere with, the conduct of the study and related procedures and/or might bias the study results interpretation, or could jeopardize the subject's safety.
• Any clinically significant findings in physical examination including vital signs.
• Neuroleptic or antidepressant (ISRS) drugs within 3 months before inclusion, except for sertraline at a maximum 100 mg/day, and stable for the last 3 months before inclusion.
• Any prescription or over-the-counter drug (except occasional use of paracetamol) in the last 2 weeks before screening.
• Patient included in a clinical study with drugs in the last three months prior to inclusion.

Typically Developing Subjects Inclusion Criteria:

• Abstinence for any drug abuse 72 hours prior to the screening visit and the observation days, except for nicotine (24 hours).
• Able to read Spanish and/or Catalan and to adhere to the study requirements.
• Signed informed consent prior to any study-mandated procedure.

Typically Developing Subjects Exclusion Criteria:

• Life-time clinically significant cardiovascular, renal, pulmonary, hepatic, onco-haematological, endocrine, gastrointestinal, mental or neurological disease.
• Any other clinically relevant concomitant disease or condition or finding at screening that in the judgment of the investigator could interfere with, the treatment thereof might interfere with, the conduct of the study and related procedures and/or might bias the study results interpretation, or could jeopardize the subject's safety.
• Any clinically significant findings in physical examination including vital signs.
• Neuroleptic drugs within 3 months prior to inclusion.
• Any prescription or over-the-counter drug (except occasional use of paracetamol) in the last 2 weeks before screening.
• Patient included in a clinical study with drugs in the last three months before inclusion.
• Personal history of infantile spasms/convulsions/epilepsy, severe head trauma or CNS infections (e.g. meningitis), except for infantile febrile seizures.
• Subjects with a current diagnosis including autism spectrum disorder or any primary psychiatric diagnosis according to DSM-5 (Diagnostic and Statistical Manual of Mental Disorders-DSM-5). Diagnoses that are secondary, such as attention deficit hyperactivity disorder, depressive disorders and conduct disorders are allowed as long as they are considered to not interfere with study conduct and are stable during the 3 months before inclusion. Related allowed treatments must be on stable dosing for the last 3 months.
• Substance use disorder according to the DSM-5 criteria except for mild alcohol use disorder and/or mild or moderate nicotine use disorder.
• Positive urine test for drugs of abuse or alcohol breath test at screening.
• Epileptiform abnormalities (excluding isolated sharp waves and beyond those expected for age).
• Any life-threatening disease.

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Primary Outcome Measures

Name	Time	Method
Biorhythm characteristics of FXS patients compared to neurotypical subjects using sleep diaries	From Day 1 to Day 28 (± 3 days)	Sleep schedule will be assessed by using sleep diaries to record the quality and quantity of sleep. A sleep diary allows recording when the subject goes to bed, when the subject wakes up during the night and when the subject wakes up in the morning. This will help to understand the sleep pattern and how much sleep the subject gets. It will also show how often the subject has interrupted sleep.
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: number of days	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the number of days \[days\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: percentage of wear	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the percentage of wear \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: total steps	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the total steps \[steps\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: steps per day	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the steps per day \[steps/day\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: average of energy expenditure	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the average of energy expenditure \[kcal/day\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: percentage of sedentary physical activity	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the percentage of sedentary physical activity \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: percentage of light physical activity	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the percentage of light physical activity \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: percentage of moderate physical activity	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the percentage of moderate physical activity \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: percentage of vigorous physical activity	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the percentage of vigorous physical activity \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: percentage of very vigorous physical activity	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the percentage of very vigorous physical activity \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: bedtime	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the bedtime \[hh:mm\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: wake-up time	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the wake-up time \[hh:mm\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: mean time in bed	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the mean time in bed \[minutes\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: total mean sleep time	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the total mean sleep time \[minutes\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: sleep efficiency	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the sleep efficiency \[%\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: wake after sleep onset	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the wake after sleep onset \[minutes\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: average of awakening	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the average of awakening \[awakes\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects using actigraphy: hours spent awake at night	From Day 1 to Day 28 (± 3 days)	Sleep-wake rhythms will be assessed using a wearable actigraph which will register the hours spent awake at night \[minutes\].
Cognitive profile of FXS patients compared to neurotypical subjects using the NIH-TCB-ID Toolbox	Day 1 and Day 28 (± 3 days)	The NIH-Toolbox Cognitive Battery for Intellectual Disabilities (NIH-TCB-ID) provides a reliable, validated and standardized measure (composite scores) that can be used as efficacy endpoints in clinical studies in patients with neurodevelopmental disorders from 3 to 85 years old. The Fluid Cognition composite score of the NIH Toolbox cognitive battery combines the scores of five tests assessing the following cognitive domains: i) Cognitive flexibility, ii) Inhibitory control and visual attention, iii) Episodic memory, iv) Processing speed and v) Working memory.
Neural oscillations using Electroencephalography (EEG)	Day 1	Multichannel EEG will be recorded using a mobile wireless helmet for high precision EEG monitoring. In total, 20 EEG channels will be used to capture brainwave activity.; The EEG test contains three well differentiated sections: auditory oddball, resting-state (eyes open and eyes closed) and auditory steady-state response (ASSR). Before starting the auditory oddball, the subject will conduct a training section (40 seconds) to present the type of sounds and to check that they have understood the task by practicing it.
Point of gaze using eye tracking	Day 1 and Day 28 (± 3 days)	Eye tracking technology will be used to record X and Y coordinates of eye position and pupil diameter. Stimuli will consist on 60 coloured photographs of adult human faces (equal numbers of males and females; different races and ethnicities), each face exhibiting a calm, happy, or fearful expression, and 60 scrambled versions of the face images. Testing will be conducted in a quiet room with the lights turned off. Each trial will start with presentation of a scrambled face image for 1 s followed immediately by its matched face image for 3 s. An inter-trial interval (ITI) containing a uniform grey screen will be shown for 0.5, 1, or 2 s, randomly determined. The order of face presentation will be pseudorandomized and each eye tracking session will last approximately 6 min.
Behavior troubles of the FXS patients compared to neurotypical subjects using the Aberrant Behavior Checklist (ABC)	≤ 1 month prior to Day 1	Standardized rating scale used for assessing problematic behavior of individuals with developmental disabilities. The questionnaire explores problem behaviors across 5 domains: Irritability (15 items), Lethargy/Socially Withdrawal (16 items), Stereotypy Behavior (7 items), Hyperactivity/Noncompliance (16 items), Inappropriate Speech (4 items). Each item is scored as 0 (never a problem), 1 (slight problem), 2 (moderately serious problem), or 3 (severe problem). For the present study the total score for each of the 5 domains or subscales will be calculated and used for the analyses, as a good measure of the psychiatric symptom and behavioral disturbance profile. In all cases, higher scores in the mentioned subscales and total score indicated a greater presence and severity of behavioral problems: Agitation \[0-45\]; Lethargy/Social Withdrawal \[0-48\]; Stereotypic Behavior \[0-21\]; Hyperactivity/Noncompliance \[0-48\] and Inappropriate Speech \[0-12\].
Adaptative functioning using the Vineland Adaptive Behaviour Scale (VABS-3)	≤ 1 month prior to Day 1	The Vineland Adaptative Behaviour Scale 3 (VABS-3) is a psychometric instrument used in child and adolescent clinical psychology for the assessment of individuals with different types of developmental delays, regarding an adaptive level of functioning by standardized interview of the person or their caregiver through their activities of daily living such as walking, talking, getting dressed, going to school, preparing a meal, etc. Three domains explore communication, socialization and daily living, which correspond to the 3 domains of adaptive functioning recognized by the American Association on Intellectual and Developmental Disabilities.
Clinician-rated global functioning using the Clinical Global Impression (CGI)	≤ 1 month prior to Day 1 and Day 28 (± 3 days)	The Clinical Global Impression Scale (CGI) comprises two companion one-item measures evaluating the following: 1. Severity of Psychopathology (Clinical Global Impression-Severity (CGI-S) Scale): establishes the baseline illness status and rates illness severity on a 7-point scale \[range 0 to 7\]: 1 for not at all ill; 2 for borderline illness; 3 for mild illness; 4 for moderately ill; 5 for very ill; 6 for severely ill; 7 among the most severe patients; and; 2. Change from the Initiation of Treatment (Clinical Global Impression-Improvement (CGI-I) Scale): rates how much the subject's illness has improved or worsened relative to the baseline state (CGI-S) on a 7-point scale \[range -7 to 7\]: 1 = very much improved; 2 = much improved; 3 = minimally improved; 4 = no change; 5 = minimally worse; 6 = much worse; and 7 = very much worse.
Stress biomarkers of the FXS patients compared to neurotypical subjects measured by cortisol concentrations in hair (occipital area)	≤ 1 month prior to Day 1	Hair cortisol analysis characterizes chronic stress as a risk factor for chronic illness progression and is known as a biomarker of the effectiveness of stress reduction interventions. Approximately 3 cm of occipital hair will be collected at the Screening visit for the determination of cortisol concentrations \[range 2.5 - 97.5 pg/mg\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects determining vitamin D (25-Hydroxy) concentrations in serum	Day 1 and Day 28 (± 3 days)	Vitamin D concentrations are also useful due to lack of sunlight exposure is the primary reason for the worldwide epidemic of vitamin D deficiency. The lack of sunlight exposure is involved in serotonin and melatonin production. Venous blood samples will be collected by individual venepuncture or via an indwelling catheter to measure vitamin D (25-Hydroxy) concentrations in serum \[25 - 150 ng/mL\].
Biorhythm characteristics of FXS patients compared to neurotypical subjects determining 24h urine cortisol concentrations	Day 1 and Day 28 (± 3 days)	The activation of the Hypothalamic Pituitary Adrenal (HPA) axis leads to the synthesis and release of cortisol, a glucocorticoid hormone that peaks in the early morning hours. 24-hour urine samples will be collected before Visit 1 and Visit 2 for the determination of cortisol concentrations \[range 11.5 - 102.0 µg/24h\].
Quality of life of the FXS patients compared to neurotypical subjects using the Paediatric Quality of Life Inventory (PedsQL)	≤ 1 month prior to Day 1	Health-related quality of life (HRQoL) is a multidimensional concept involving physical, psychological, social, and cognitive aspects of life. Individuals with Fragile X syndrome (FXS) experience a life-long disorder that impacts the HRQoL of the affected individual and their family. HRQoL has been correlated with established measures of functioning in FXS using the: i) Cognitive Functioning Scale \[0-100\], ii) Quality of Life Scale, which is composite of the following items: Physical health and activities \[0-32\], Emotional state \[0-20\], Social activities \[range 0-20\], School activities \[0-20\]; and iii) Family impact scale, which is composite of the following items: Physical health and activities \[0-24\], Emotional state \[0-20\], Social activities \[0-16\], Cognitive function \[0-20\], Communication \[0-12\], Preoccupation \[0-20\], Daily activities \[0-12\] and Family relationships \[0-20\].
Quality of sleep of the FXS patients compared to neurotypical subjects using Pittsburgh sleep quality index (PSQI)	Day 1 and Day 28 (± 3 days)	The Pittsburgh sleep quality index (PSQI) consists in a self-report questionnaire that assesses sleep quality and quantity. The 19-item self-report questionnaire yields 7 component scores: i) subjective sleep quality \[0-3\], ii) sleep latency \[0-3\], ii) duration of the sleep \[0-3\], iii) habitual sleep efficiency \[0-3\], iv) sleep disturbances \[0-3\], v) use of sleeping medication \[0-3\], and vi) daytime dysfunction \[0-3\]. There are five additional questions that are completed by a bed partner if there is one.
FMRP protein in peripheral blood of FXS patients in peripheral blood compared to neurotypical subjects	≤ 1 month prior to Day 1	Venous blood samples will be obtained by extraction of peripheral blood from participants in a EDTA tube. The levels of FMRP in the FMR1 gene will be reported as a relative value of the mean levels calculated for control \[range 0-1\].
miRNA profile of FXS patients in plasma compared to neurotypical subjects	Day 1 and Day 28 (± 3 days)	Venous blood samples will be obtained by extraction of peripheral blood from participants. For the determination of the miRNomic profile, 200 μL of human plasma will be processed using Plasma/Serum miRNeasy Serum/Plasma kit to extract RNA enriched in small RNAs.

Trial Locations

Locations (2)

Consorci Corporació Sanitaria Parc Taulí. Institut Investigació i Innovació Parc Taulí (I3PT); 🇪🇸 Sabadell, Barcelona, Spain

Hospital del Mar Research Institute (HMRI); 🇪🇸 Barcelona, Spain