Objective Purpose of the study was to identify the relationship among actual plasmatic levels of steroid hormones and behavioral manifestations in boys with autism and to assess the genetic contribution to these manifestations.Methods 172 boys with autism under 10 years of age and 135 neurotypical boys attended the study. ADI-R and ADOS-2 were used to evaluate the core symptom severities. Problem behavior was assessed using BPI-01 questionnaire. Levels of testosterone, estradiol, dehydroepiandrosterone, dehydroepiandrosterone-sulfate and sex hormone binding globulin (SHBG) were measured in plasma of autistic boys. Three SNPs (in ESR1, SHBG, SRD5A2 genes) and one STR in AR gene (number of CAG repeats in first exon) were assessed. Hormonal levels and number of CAG repeats in AR gene were used for correlation analysis with behavioral measures. Genotype and allelic frequencies were compared among autistic and neurotypical boys.Results We found negative relationship among SHBG levels and restricted, repetitive behaviors (measured by ADOS-2) and positive relationship among actual testosterone levels and frequency of stereotyped behavior (measured by BPI-01).Conclusion Actual levels of SHBG and testosterone are related to severities of restricted and repetitive behaviors in boys with autism. Mechanisms of action of these hormones in brain require further investigation.

Background: Autism spectrum disorder (ASD) is a heterogeneous group of neurodevelopmental disorders characterized by a complex, multifactorial etiology with a strong genetic contribution. Our study aimed to evaluate the link between the burden of rare genetic variants within a specific panel of ASD and intellectual disability-associated genes and phenotypic variability in a cohort of children with autism in Slovakia. Methods: Gene burden scores were calculated based on pathogenic, likely pathogenic, and uncertain significance rare DNA variants identified by whole-exome sequencing. We then assessed the effect of three different scoring methods on the variance across 15 psycho-behavioral parameters describing the phenotypic profiles of 117 ASD probands. Results: The burden score showed a significant multivariate effect on the combination of psycho-behavioral parameters. This score was associated with the social affect of ADOS-2, as well as with the socialization domain, and total adaptive behavior scores from the Vineland Adaptive Behavior Scales-3 (VABS). While a score based solely on count of pathogenic and likely pathogenic variants did not show a multivariate effect, incorporating variants of uncertain significance revealed a multivariate effect on two adaptive behavior parameters: daily living skills and total adaptive behavior score (VABS). Conclusions: Our findings partially explain the variability in phenotypic manifestation in our ASD patient cohort, highlighting the importance of considering the cumulative effect of rare genetic variants, including those of uncertain significance, in shaping the diverse clinical presentation of ASD.

In this article, we present a case study of a five-year-old girl with autism and developmental delay, conducted at the Academic Center for Autism Research in Bratislava, Slovakia. The girl was diagnosed using Autism Diagnostic Observation Schedule-Second Edition (ADOS-2) and Autism Diagnostic Interview-Revised (ADI-R) instruments and met the criteria for autism spectrum disorder. Intellectual functioning was in the markedly below-average range, as indicated by the Snijders-Oomen Nonverbal Intelligence Test-Revised (SON-R) examination, and her level of adaptive functioning was significantly reduced. Neurological signs included atypical leukoencephalopathy, hypotonia, sensorineural hearing loss, and sleep disturbances. The patient underwent genetic testing, including microarray-based copy number variation (CNV) detection, which yielded negative results. However, whole exome sequencing (WES) analysis pointed out a damaging homozygous variant in the EIF3F (Eukaryotic Translation Initiation Factor 3 Subunit F) gene, confirming the diagnosis of intellectual developmental disorder autosomal recessive 67. Segregation analysis in the family revealed that the asymptomatic parents were carriers of the pathogenic variant in EIF3F. Our study contributes to the phenotypic profiling of this rare syndromic neurodevelopmental disorder and points out the irreplaceability of WES analysis in genetic diagnostics of autism and developmental delay. This appeals to the need for WES to be accepted as a routine diagnostic tool in Slovakia.

Here, we present a case study of twin boys aged 2 and 7 years who both met the diagnostic criteria for autism spectrum disorders (ASDs) based on the standard diagnostic instruments ADOS‐2 and ADI‐R. The clinical indication for genetic diagnostics in the first boy was autism with high severity of symptoms, delayed speech development, and mild facial dysmorphia. The second boy’s indication was autism with moderate severity of symptoms, delayed speech development, mild facial features, slowed psychomotor development, and microcephaly. The microarray‐based analysis of chromosome aberrations revealed a heterozygous 977,456 bp deletion of region 10q26.3 in both boys. The region includes 28 genes, some of these genes are important in the development of the central nervous and urogenital systems, and heterozygous deletions in this region have been associated with mental retardation, growth and development disorders, and craniofacial anomalies. The whole exome sequencing confirmed the presence of this deletion in both boys and, at the same time, led to the identification of a pathogenic SNV variant in the TRIO gene in the boy with microcephaly and delayed psychomotor development, which may explain the different phenotype of both boys. However, the segregation analysis of these variants in the family revealed that the microdeletion was inherited from the asymptomatic father, and the c.2149C > T variant in the TRIO gene was inherited from the asymptomatic mother, making the diagnostic finding uncertain. This case highlights that when pathogenic or likely pathogenic variants are inherited from unaffected parents, the clinical phenotype may result from a combined burden of multiple rare variants and polygenic risk, underscoring the importance of a comprehensive genomic analysis in complex cases. Thus, we emphasize the importance of utilizing available methods, such as whole exome sequencing besides microarray‐based comparative genomic hybridization, in the genetic diagnosis of autism patients in Slovakia.