Neuralized1 and RGS14 Genes

Study Overview
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Brief Summary
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Autism is a broad spectrum neurodevelopmental disease. Some individuals with ADS by high cognitive functions are diagnosed with High Functioning Autism (HFA). In some studies, it has been shown that NEURL1 gene increases learning and memory and RGS14 gene is suppressed them. We aimed to evaluate the differences between the expression levels of these genes between ASD, HFA and healthy controls and the role of these genes in the pathogenesis of ASD. Patients with 20 ASD and 20 HFA, and 20 healthy controls compatible with patient ages were included in this study. Expression of NEURL-1 and RGS14 genes was evaluated by quantitative Real Time PCR (qRT-PCR).

Detailed Description
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ASD is a neurological disease starting in the early stages of life and is characterized by cognitive and behavioral disorders (Ansel et al., 2008;Alvares et al., 2020). It is considered that the etiology of ASD stems from genetic, epigenetic and environmental factors; however, it has not yet been definitively clarified(Ito et al., 2017). we aimed to evaluate the differences between the expression levels of these genes between ASD, HFA and healthy controls and the role of these genes in the pathogenesis of ASD. Method: Patients with ASD (n=20) and HFA (n=20), and healthy controls (n=20) compatible with patient ages were included in this study. Clinical evaluations of the patients were made and classification was made in accordance with DSM-IV diagnostic criteria. High Pure RNA Isolation Kit (Roche Diagnostic, Version 12, Germany) was used for RNA isolation. cDNA synthesis was performed from these RNAs with the ranscriptor High Fidelity cDNA Synthesis Kit (Roche Diagnostics, GmbH, Mannheim). qRT-PCR was performed using the LightCycler®480 Real Time Ready Assay Master Probe Kit (Roche Diagnostics, GmbH, Mannheim).The incubation was made with the PCR device program for 10 minutes at 95oC for 45 cycles, for 10 sec at 95oC, and for 60 sec at 60oC. The Ct values were obtained from the Light Cycler 480 Software Program, and both genes were analyzed separately. The comparative CT method (2-ΔΔCT) was used to determine the relative quantification of target genes, normalized to a housekeeping gene (β-actin). Statisticaly: The results of the experiments were evaluated using R 3.1.1 (www.r-project.org). and Chi-Square Tests, Mann-Whitney U-Test, Kruskal-Wallis H-Tests. The P<0.05 level was taken as significant.

Official Title
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Role of Neuralized1 and RGS14 Genes With ASD Patients

Conditions
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Healthy, Autism Spectrum Disorder, High-functioning Autism

Intervention / Treatment
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* Diagnostic Test: Neurl1 gene expression
* Diagnostic Test: RGS14 gene expression


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Being diagnosed ASD or HFA patient, Being between the ages of 2-16. Exclusion Criteria: To use medicine, Have a other syndromic illness, Being younger than 2 years old or over 16 years old.

Ages Eligible for Study
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Minimum Age: 2 Years
Maximum Age: 16 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Patients with Autism Spectrum Disorder (ASD)<br>Patients diagnosed with ASD in a child psychiatry clinic. | Diagnostic Test: Neurl1 gene expression<br>* This observational case control study. The gene expression was examined.<br>Diagnostic Test: RGS14 gene expression<br>* This observational case control study. The gene expression was examined.<br>|
| Patients with High Functioning Autism (HFA)<br>Patients diagnosed with ASD in a child psychiatry clinic and with an IQ above 70. | Diagnostic Test: Neurl1 gene expression<br>* This observational case control study. The gene expression was examined.<br>Diagnostic Test: RGS14 gene expression<br>* This observational case control study. The gene expression was examined.<br>|
| Healty control<br>Healthy volunteers who are in the age range compatible with the patient groups. | Diagnostic Test: Neurl1 gene expression<br>* This observational case control study. The gene expression was examined.<br>Diagnostic Test: RGS14 gene expression<br>* This observational case control study. The gene expression was examined.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| NEURL1 gene expression levels | After RNA isolation from blood samples of the subjects, NEURL1 gene expression was studied by QPCR method. The 2-ΔΔCT method was applied for the relative quantification of the samples that were normalized with ACTB. | Two months | 
| RGS14 gene expression levels | After RNA isolation from blood samples of the subjects, RGS14 gene expression was studied by QPCR method. The 2-ΔΔCT method was applied for the relative quantification of the samples that were normalized with ACTB. | Two months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Age | Age of subjects | an average of 1 year | 
| Gender | Gender (male/female) of subjects | an average of 1 year | 
| Intellectual disability (ID) | Intellectual disability is when a person has certain limitations in cognitive functioning and skills, including communication, social and self-care skills.It was determined according to DSM-IV diagnostic criteria and clinical evaluation. | an average of 1 year | 
| Consanguinity | Relationships of consanguinity between subjects were evaluated in terms of pathogenesis of the disease. | an average of 1 year | 
| Presence of Neurological Disease in Relatives | In the presence of a neurological disease in relatives, its relationship with the pathogenesis of the disease was evaluated. | an average of 1 year | 
| Corelation tests | The relationships of the clinical and demographical findings in the study groups with the genes were evaluated statistically. | an average of 1 year | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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ASD, High Functioning Autism, NEURL1, RGS14, Gene expression