Sains Malaysiana 49(3)(2020): 635-642

http://dx.doi.org/10.17576/jsm-2020-4903-18

DNA Methylation Analysis of AKT1 Promoter and HTR2A Exon-I of Malaysian Schizophrenia Multiplex Families with Lower Cognitive Performance

(Analisis Metilasi DNA Promoter AKT1 dan Exon-I HTR2A pada Keluarga Skizofrenia di Malaysia dengan Prestasi Kognitif Rendah)

ERNA LAERE¹, TZE JEN CHOW², PEK YEE TANG², SIEW YIM LOH³, HOI SEN YONG⁴, ABDUL KADIR, ABU BAKAR⁵ & SHIAU FOON TEE¹*

¹Lee Kong Chian, Faculty of Engineering and Science, Department of Chemical Engineering, Universiti Tunku Abdul Rahman, Jalan Sungai Long, Bandar Sungai Long, Cheras 43000 Kajang, Selangor Darul Ehsan, Malaysia

²Lee Kong Chian, Faculty of Engineering and Science, Department of Mechatronics and Biomedical Engineering, Universiti Tunku Abdul Rahman, Jalan Sungai Long, Bandar Sungai Long, Cheras 43000 Kajang, Selangor Darul Ehsan, Malaysia

³Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

⁴Institute of Biological Sciences, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

⁵Jalan Persiaran Kempas Baru, 81200 Johor Bahru, Johor Darul Takzim, Malaysia

Diserahkan: 2 Oktober 2018/Diterima: 6 Mac 2019

ABSTRACT

Dysfunction of cognitive performance in schizophrenia has been associated with aberrant alteration of DNA methylation of several schizophrenia-risk genes. AKT1 and HTR2A are among the candidate genes for schizophrenia. Their expressions were found reduced in schizophrenia patients. Thus, we aimed to study the methylation status of AKT1 promoter and HTR2A exon-I in Malaysian schizophrenia patients and their affected family members. In this study, each participant was required to perform Trail Making Test (TMT) part A and B to measure their cognitive performance. Genomic DNA extracted from the peripheral blood of 12 Malaysian schizophrenia families and 12 controls families, was subjected to bisulfite conversion. The methylation status of CpG sites of AKT1 promoter at Chr14: 104796054 and HTR2A exon-I at Chr13: 46896918 were identified using methylation-specific polymerase chain reaction (MSP). Our results showed that schizophrenia patients performed worse in both TMT-A and B (p<0.0001) than healthy controls. The patients also displayed significantly (p=0.023) high level of methylation in AKT1 promoter compared to controls. Meanwhile, no significant difference (p=0.248) in methylation status was observed in HTR2A exon-I between schizophrenia and control groups. Therefore, methylation of AKT1 promoter in peripheral bloods of patients may involve in cognitive impairment and schizophrenia pathology. In addition, we were able to demonstrate the heritability of DNA methylation status across family members.

Keywords: Cognitive performance; DNA methylation; schizophrenia; Trail-Making Test

Abstrak

Disfungsi prestasi kognitif dalam skizofrenia telah dikaitkan dengan perubahan metilasi DNA pada beberapa gen yang berisiko menyebabkan skizofrenia. AKT1 dan HTR2A adalah antara calon gen kepada skizofrenia. Ekspresi gen-gen tersebut didapati berkurangan pada pesakit skizofrenia. Oleh itu, kertas ini bertujuan untuk mengkaji status metilasi promoter AKT1 dan exon-I HTR2A pada pesakit skizofrenia di Malaysia dan juga ahli keluarga mereka yang menghidap skizofrenia. Dalam kajian ini, setiap peserta dikehendaki melakukan Ujian Membuat Jejak (TMT) bahagian A dan B untuk mengukur prestasi kognitif mereka. DNA genom yang telah diekstrak daripada darah periferal 12 keluarga skizofrenia di Malaysia dan 12 keluarga kawalan sihat, tertakluk kepada penukaran bisulfite. Status metilasi tapak CpG dalam promoter AKT1 di Chr14: 104796054 dan dalam exon-I HTR2A di Chr13: 46896918 dikaji dengan menggunakan metilasi-khusus tindak balas rantaian polimerase (MSP). Keputusan menunjukkan bahawa pesakit skizofrenia mendapat keputusan yang teruk dalam kedua-dua bahagian TMT-A dan B (p<0.0001) berbanding dengan kawalan sihat. Pesakit skizofrenia juga menunjukkan tahap metilasi promoter AKT1 yang tinggi secara signifikan berbanding dengan kawalan (p=0.023). Sementara itu, tiada perbezaan yang signifikan (p=0.248) diperhatikan dalam metilasi status exon-I HTR2A antara kumpulan skizofrenia dan kawalan sihat. Kertas ini menunjukkan bahawa metilasi promoter AKT1 dalam darah periferal pesakit skizofrenia mungkin terlibat dalam kecacatan kognitif dan patologi skizofrenia. Tambahan pula, kami dapat menunjukkan bahawa kewarisan status metilasi DNA dalam kalangan ahli keluarga.

Kata kunci: Metilasi DNA; prestasi kognitif; skizofrenia; Ujian Membuat Jejak

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*Pengarang untuk surat-menyurat; email: teesf@utar.edu.my