Assessment of miR-181b-5p, miR-23a-3p, BCL-2, and IL-6 in Peripheral Blood Mononuclear Cells of Autistic Patients; Likelihood of Reliable Biomarkers
Autism is a neurodevelopmental disorder that is recognized by stereotypic and repetitive behaviors after 2 years of old. Dysregulation of the immune system, especially inflammation which is mostly regulated by IL-6, imposes a deficit in CNS development. Along with this crucial biomarker, researchers have proposed BCL-2, micro RNA-23a-3p (miR-23a-3p), miR-181b-5p as other probable biomarkers involved in inflammation and apoptosis. The aim of the study was to evaluate the alteration in the expression of these biomarkers in a group of autism spectrum disorder (ASD) children. Peripheral blood mononuclear cells (PBMCs) were obtained from 37 autistic patients. After RNA extraction with precipitation method, the Syber green qReal-time Polymerase Chain Reaction (PCR) was performed in order to evaluate the possible alteration in the expression of IL-6, BCL-2, miR-181b-5p, and miR-23a-3p. The results were compared with healthy controls. IL-6 was significantly upregulated in ASD patients (p=0.003). On the other hand, miR-23a was upregulated and BCL-2 downregulated in ASD patients but the changes were not significant. In initial evaluations, expression changes of miR-181b-5p were not statistically significant. However, when Patients were divided into two groups of upregulated and downregulated, re-evaluation showed that both up- (p=0.005) and down-regulation (p=0.004) (i.e. changes regardless of the direction) of miR-181b were significant in autistic children. IL-6 and miR-181b-5p can have proper diagnostic values and are reliable biomarkers with high sensitivity and specificity. On the other hand, PBMC can be utilized for such studies and also evaluation of patients' condition instead of brain tissue as it is less accessible.
2. Kordulewska NK, Kostyra E, Chwała B, Moszyńska M, Cieślińska A, Fiedorowicz E, et al. A novel concept of immunological and allergy interactions in Autism Spectrum Disorders: Molecular, anti-inflammatory effect of osthole. Int Immunopharmacol2019;72:1-11.
3. Mehler MF, Purpura DP. Autism, fever, epigenetics and the locus coeruleus. Brain Res Rev 2009;59(2):388-92.
4. Kałużna-Czaplińska J, Żurawicz E, Jóźwik-Pruska J. Focus on the Social Aspect of Autism. Journal of autism and developmental disorders. 2018;48(5):1861-7.
5. Ashwood P, Wills S, Van de Water J. The immune response in autism: a new frontier for autism research. J Autism Dev Disord 2006;80(1):1-15.
6. Elder JH, Kreider CM, Brasher SN, Ansell M. Clinical impact of early diagnosis of autism on the prognosis and parent–child relationships. Psychol Res Behav Manag 2017;10:283-92.
7. Masi A, Glozier N, Dale R, Guastella AJ. The immune system, cytokines, and biomarkers in autism spectrum disorder. Neurosci Bull 2017;33(2):194-204.
8. Meltzer A, Van de Water J. The role of the immune system in autism spectrum disorder. Neuropsychopharmacology. 2017;42(1):284-98.
9. Goines PE, Ashwood P. Cytokine dysregulation in autism spectrum disorders (ASD): possible role of the environment. Neurotoxicol Teratol 2013;36:67-81.
10. Morgan JT, Chana G, Pardo CA, Achim C, Semendeferi K, Buckwalter J, et al. Microglial activation and increased microglial density observed in the dorsolateral prefrontal cortex in autism. Biol Psychiatry 2010;68(4):368-76.
11. Malik M, Sheikh AM, Wen G, Spivack W, Brown WT, Li X. Expression of inflammatory cytokines, Bcl2 and cathepsin D are altered in lymphoblasts of autistic subjects. Immunobiology 2011;216(1-2):80-5.
12. Wei H, Alberts I, Li X. The apoptotic perspective of autism. Int J Dev Neurosci 2014;36:13-8.
13. Wei H, Malik M, Sheikh AM, Merz G, Brown WT, Li X. Abnormal cell properties and down-regulated FAK-Src complex signaling in B lymphoblasts of autistic subjects. Am J Pathol 2011;179(1):66-74.
14. Filipowicz W, Bhattacharyya SN, Sonenberg NJNrg. Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight? Nat Rev Genet2008;9(2):102-14.
15. Pountos I, Corscadden D, Emery P, Giannoudis PVJI. Mesenchymal stem cell tissue engineering: techniques for isolation, expansion and application. Injury2007;38:S23-S33.
16. Weber JA, Baxter DH, Zhang S, Huang DY, Huang KH, Lee MJ, et al. Clin Chem 2010;56(11):1733-41.
17. Kichukova TM, Popov NT, Ivanov HY, Vachev TI. Circulating microRNAs as a novel class of potential diagnostic biomarkers in neuropsychiatric disorders. Folia Med (Plovdiv) 2015;57(3-4):159-72.
18. Sheinerman KS, Umansky SR. Circulating cell-free microRNA as biomarkers for screening, diagnosis and monitoring of neurodegenerative diseases and other neurologic pathologies. Front Cell Neurosci 2013;7:150.
19. Vasu MM, Anitha A, Thanseem I, Suzuki K, Yamada K, Takahashi T, et al. Serum microRNA profiles in children with autism. MolAutism 2014;5(1):40.
20. Talebizadeh Z, Butler MG, Theodoro MF. Feasibility and relevance of examining lymphoblastoid cell lines to study role of microRNAs in autism. Autism Res 2008;1(4):240-50.
21. Seno MMG, Hu P, Gwadry FG, Pinto D, Marshall CR, Casallo G, et al. Gene and miRNA expression profiles in autism spectrum disorders. Brain Res 2011;1380:85-97.
22. Hu VW, Nguyen A, Kim KS, Steinberg ME, Sarachana T, Scully MA, et al. Gene expression profiling of lymphoblasts from autistic and nonaffected sib pairs: altered pathways in neuronal development and steroid biosynthesis. PloS one 2009;4(6):e5775.
23. Suda S, Iwata K, Shimmura C, Kameno Y, Anitha A, Thanseem I, et al. Decreased expression of axon-guidance receptors in the anterior cingulate cortex in autism. Mol Autism 2011;2(1):14.
24. Sarachana T, Zhou R, Chen G, Manji HK, Hu VW. Investigation of post-transcriptional gene regulatory networks associated with autism spectrum disorders by microRNA expression profiling of lymphoblastoid cell lines. Genome Med 2010;2(4):23.
25. Shi W, Du J, Qi Y, Liang G, Wang T, Li S, et al. Aberrant expression of serum miRNAs in schizophrenia. J Psychiatr Res 2012;46(2):198-204.
26. Ouyang Y-B, Lu Y, Yue S, Giffard RG. miR-181 targets multiple Bcl-2 family members and influences apoptosis and mitochondrial function in astrocytes. Mitochondrion 2012;12(2):213-9.
27. Gardiner E, Beveridge N, Wu J, Carr V, Scott R, Tooney P, et al. Imprinted DLK1-DIO3 region of 14q32 defines a schizophrenia-associated miRNA signature in peripheral blood mononuclear cells. Mol Psychiatry 2012;17(8):827-40.
28. Mellios N, Sur M. The emerging role of microRNAs in schizophrenia and autism spectrum disorders. Front Psychiatry 2012;3:39.
29. Tayebi B, Abrishami F, Alizadeh S, Minayi N, Mohammadian M, Soleimani M, et al. Modulation of microRNAs expression in hematopoietic stem cells treated with sodium butyrate in inducing fetal
hemoglobin expression. Artif Cells Nanomed Biotechnol2017;45(1):146-56.
30. Ashwood P, Krakowiak P, Hertz-Picciotto I, Hansen R, Pessah I, Van de Water J. Elevated plasma cytokines in autism spectrum disorders provide evidence of immune dysfunction and are associated with impaired behavioral outcome. Brain Behav Immun 2011;25(1):40-5.
31. Masi A, Breen EJ, Alvares GA, Glozier N, Hickie IB, Hunt A, et al. Cytokine levels and associations with symptom severity in male and female children with autism spectrum disorder. MolAutism2017;8(1):63.
32. Araghi-Niknam M, Fatemi SH. Levels of Bcl-2 and P53 are altered in superior frontal and cerebellar cortices of autistic subjects. Cell Mol Neurobiol 2003; 23(6):945-52.
33. Shen L, Lin Y, Sun Z, Yuan X, Chen L, Shen B. Knowledge-guided bioinformatics model for identifying autism spectrum disorder diagnostic microrna biomarkers. Sci Rep 2016; 6:39663.
34. Mandell DS, Wiggins LD, Carpenter LA, Daniels J, DiGuiseppi C, Durkin MS, et al. Racial/ethnic disparities in the identification of children with autism spectrum disorders. Am J Public Health 2009; 99(3):493-8.
35. Becerra TA, von Ehrenstein OS, Heck JE, Olsen J, Arah OA, Jeste SS, et al. Autism spectrum disorders and race, ethnicity, and nativity: a population-based study. Pediatrics 2014; 2013-3928.