Circulating Non-Coding RNAs as a Signature of Autism

Spectrum Disorder Symptomatology

Salam Salloum-Asfar , Ahmed K. Elsayed , Saba F. Elhag and Sara A. Abdulla

Abstract Results and Discussion

Autism spectrum disorder (ASD), observed i n early chil dhood dev elopm ent is a c omplex disorder ASD diagnosis – Through the study of behavi oral phenoty pes and c harac teristics, and identification of associ ated biomarkers. ncRNAs was derived from
of the neur ologic al dev elopment. The two main symptoms of ASD are res tricted, repetitiv e a small sample size of pl atel et-depleted plasma to identify potential bi omarkers. The s amples from childr en aged between 3 to 15 wer e classified into
patterns of behavior, i nter ests, or activities and deficits i n s ocial c ommunication and soci al two main groups – Individuals with ASD vs. healthy control and individuals with severe ASD vs. less severe ASD.
interacti on. Clinical diagnosis c annot be us ed standal one as it risks ineffective therapeutic
treatments due to late or missed diagnosis. It is difficult to clinically diagnose due to its compl ex
nature, but identifying bi omarkers bas ed on the s everity of ASD has becom e the route for
diagnosis in associ ation to subjectiv e and clinic al di agnosis, and allows for personalized
therapeutic i nterventions. N on-coding RNA ( ncRNA) like miRNAs, piRNAs, snoRNAs, Y-RNAs,
tRNAs, and lncRNAs pl ays a m ajor rol e i n the proper devel opment and functi oning of the c entral
nervous system and w ere previously found to play a role in neurodegener ative diseases . Hence,
the focus on circulating ncRNAs as important biomarkers for the study of ASD.

Objectives
• Amongs t the 5 bioty pes of ncRNA (miRNAs, piRNAs, snoRNAs, Y-RNAs, tRNAs, and l ncRNAs) analyzed, miR-302 from family miRNA had high read
● To use the tec hnology of next-generation sequenci ng to identify different circul ation counts and were express ed in high l evels in sev ere ASD group when compared to less sev ere. miRNA is known to negatively influenc e the neural
non-coding RNA (cir-ncRNA). development, neuronal receptors, and synaptic plasticity.
● To confirm the stable expressi on of miRNAs, piRNAs, snoRNAs, Y-RNAs, tRNAs, and
lncRNAs in the plasma. • miR-135b-5p, a ty pe of miRNA, w as also express ed in high lev els in sev ere ASD group c ompared to l ess sev ere group. Dysregulation and disrupti on
of miR-135b-5p was observed in schizophrenia 1 and is associated with neuropsychiatric behavior.

Methodology • Pathway enr ichm ent analysis showed dysregul ation i n the l evels of ex pression of ASD ass ociated molec ules like DKK1, EPH B6, NTRK3,
P53, AGO2, PTEN, and BR AF, and distortion of neuro-rel ated pathways. DKK found in the brai n and EPHB6 inv olved i n gut homeostasis are link ed
to ASD. Similarly, NTKR3 is observed in c ases of both ASD and As per gers. Altered p53 is also observ ed i n ASD. Pathway
analysis cpnfirmed that AGO3 was more specific in individuals with ASD compared to healthy cohort.

• High-throughput sequencing showed the presence of other ncRNAs like snoRNAs, Y-RNAs, tRNAs, and lncRNAs having regulatory functions.

• Small RNA high-throughput sequencing showed the pr esence of 110 nucleotides (nt) long Ro-ass ociated Y-RNAs (also c alled RNYs or Y-RNAs) in
the plasm a. Y-RNA, hY3, and pseudogene hY3P1 were downregulated, and RNY4 Ps eudogene 28 and 29 wer e differentially express ed in
individuals with severe ASD. YRNA is important for the initiation of chromosomal DNA replication, RNA stability, and cellular responses to stress

• The SNORA69 (U69) is the m ost up-regulated small nucleolar RNA, whereas SNORD42A (U42) is the most dow n-regul ated snoRNA in
individuals that expressed more severe ASD symptoms

Conclusion
The identification of ASD biomarkers and the study of their roles in the neurological devel opmental process c an help acceler ate and improve
therapeutic interventions for the diagnosis and treatment of ASD.