Selection of Suitable Reference Genes for Analysis of Salivary Transcriptome in Non-Syndromic Autistic Male Children

Panahi, Yasin; Moghaddam, Fahimeh Salasar; Ghasemi, Zahra; Jafari, Mandana Hadi; Badv, Reza Shervin; Eskandari, Mohammad Reza; Pedram, Mehrdad

doi:10.3390/ijms17101711

Cited by 8 publications

(9 citation statements)

References 63 publications

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“…All cases were collected during a three-year period from 10 different cities spanning nine provinces. Children with idiopathic autism were diagnosed as previously described (42). Briefly, each child with autism was diagnosed by at least two qualified clinicians, a child neurologist and a specialized psychiatrist, according to the Diagnostic and Statistical Manual of Mental Disorders fourth edition, Text Revised (DSM-IV-TR) criteria.…”

Section: Methodsmentioning

confidence: 99%

Sexual Dimorphism in Telomere Length in Childhood Autism

Panahi

Moghaddam²,

Babaei

et al. 2020

Preprint

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Autism spectrum disorders (ASD) are lifelong heterogeneous set of neurodevelopmental conditions with strikingly profound male prevalence. Differences in sex biology and hormones are thought to play key roles in ASD prevalence and outcome, but the underlying molecular mechanisms responsible for ASD sex-differential risk are not well understood. Two recent studies reported a significant association between shortened telomere length (TL) and autistic children. However, the role of gender bias has been overlooked. Here, we carefully examined the status of average TL among non-syndromic male and female children with autism, and we also took a close look at the data from earlier reports. A total of 58 children were recruited for this project, including 24 apparently non-syndromic autistic children (14 males and 10 females), their healthy siblings (n = 10), and 24 sex-, age, and location-matched healthy controls. Relative TLs (RTL) were assessed by the monochrom multiplex quantitative polymerase chain reaction (MMQPCR) technique, using genomic DNA extracted from saliva samples. Data analysis showed that gender and age had strong impacts on average RTLs among the study groups. In a sex stratified manner, autistic male children had significantly shorter average RTL than their female counterparts. Only male children with autism showed a homogeneous pattern of shorter RTLs compared with their respective healthy controls. Our findings are indicative of a sexually dimorphic pattern of TL in childhood autism. The data presented here have important implications for the role of telomere biology in the molecular mechanisms responsible for ASD male bias prevalence and etiology.

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Section: Methodsmentioning

confidence: 99%

Sexual Dimorphism in Telomere Length in Childhood Autism

Panahi

Moghaddam²,

Babaei

et al. 2020

Preprint

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“…Our observations on the sexually dimorphic patterns of telomere length in children with nonsyndromic autism [33] Saliva sampling was done using the Oragene collection kits (DNA genoTeK Inc., Ottawa, Ontario, Canada) as described previously [35]. OG-250 saliva-collection kits were used for genomic DNA (gDNA) extraction, and RE-100 kits were used for RNA extraction.…”

Section: Discussionmentioning

confidence: 99%

“…Total RNA was extracted from whole saliva as described in detail previously [35]. Briefly, total RNA was reverse transcribed to Complementary DNA (cDNA) by using the PrimeScriptTM RT reagent Kit (TaKaRa Inc., Shiga, Japan) following the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

“…We have previously reported a detailed evaluation of GAPDH and YWHAZ as appropriate combined reference genes for normalization of RT-qPCR data for saliva transcript level analysis in autistic male children [35]. We performed a similar systematic two-phased analysis among our autistic and contorl female samples.…”

Section: Differential Terra Expression In Male Children With Autismmentioning

confidence: 99%

“…Selection and validation of GAPDH and YWHAZ combination as the most stable and suitable reference genes for analysis of the salivary transcriptome in male children with non-syndromic autism has been described previously [35]. A similar analysis of the cDNAs derived from female saliva samples showed that although GAPDH and YWHAZ were still the best among a panel of nine candidate reference genes examined, they would not be sufficient for proper normalization of the RT-qPCR data obtained from female subjects ( Supplementary Material, Fig.…”

Section: Rna Extraction and Analysis Of Terra Transcript Levels With mentioning

confidence: 99%

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Differential Levels of Telomeric Oxidized Bases and TERRA Transcripts in Childhood Autism

Eftekhar

Panahi

Moghaddam

et al. 2020

Preprint

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The underlying molecular mechanisms responsible for the etiology of autism and its sex-biased prevalence remain largely elusive. Abnormally shortened telomeres have recently been associated with autism. We have previously shown that children with non-syndromic autism exhibit a sexually dimorphic pattern of relative telomere length (RTL). Only male children with autism have significantly shorter RTLs than the healthy controls and paired siblings. Autistic females have substantially longer RTLs than autistic males. Aberrantly high levels of oxidative stress plays a fundamental role in the pathophysiology of autism, and telomeres are thought to be susceptible to oxidative damage due to their high guanine-repeat content. Employing a quantitative PCR (qPCR)-based method, telomeric oxidized base lesions were measured using genomic DNA extracted from saliva samples, and levels of telomeric RNA transcripts know as TERRA were evaluated using reverse transcriptase qPCR technique. Our data show that the autistic children exhibit substantially higher levels of oxidative base lesions at their telomeres than the healthy controls and paired siblings. Intriguingly, despite having significantly longer RTLs, female children with autism have even higher levels of telomeric oxidized bases than their male counterparts. Furthermore, despite having significantly shorter RTLs, the male children with autism exhibit lower levels of TERRA expression from the short arms of chromosomes 17 and X/P compared to their individually-matched healthy controls. These findings open a fresh angle into autism. Abnormal TL and high levels of telomeric oxidized bases may serve as biomarkers for childhood autism.

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