ﺷﻨﺎﺳﺎﯾﯽ ﺟﻬﺶﻫﺎي ﻣﺴﺒﺐ ﺑﯿﻤﺎري در ژنﻫﺎي ﻣﺮﺗﺒﻂ ﺑﺎ ﺑﯿﻤﺎري اﭘﯿﺰودﯾﮏ ﮐﻤﺎ در ﯾﮏ ﺧﺎﻧﻮاده اي ﺑﺎ ﺳﻪ دﺧﺘﺮ ﻣﺒﺘﻼ ﺑﻪ اﯾﻦ ﺑﯿﻤﺎري ﺑﺎ ﮐﺎرﺑﺮد روشﻫﺎي ﺗﻮاﻟﯽ ﯾﺎﺑﯽ ﻧﺴﻞ ﺟﺪﯾﺪ

The identification of the disease -causing mutations in genes associated with episodic coma in a family with three girls affected with this disorder using Next Generation Sequencing (NGS)

ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ ﯾﮏ ﻣﻮرد ﮐﻤﺎي ﺗﮑﺮار ﺷﻮﻧﺪه ﺧﺎﻧﻮادﮔﯽ را ﮔﺰارش ﻣﯽﮐﻨﺪ ﮐﻪ در آن ﺳـﻪ دﺧﺘـﺮ ﺗﺸﻨﺠﺎت ﺳﺮﮐﺶ ﻧﺸﺎن ﻣﯽ دادﻧﺪ ﮐﻪ ﻣﻨﺠﺮ ﺑﻪ ﮐﻤﺎ ﻣﯽﺷـﺪ . ﭘﻨـﻞ ژﻧـﯽ ﻫﺪﻓﻤﻨـﺪ ﺑـﺮاي ﺻـﺮع ﺑـﺎ ﮐـﺎرﺑ ﺮد ﺗﻮاﻟﯽﯾﺎﺑﯽ ﻧﺴﻞ ﺟﺪﯾﺪ ﺑﻪﮐﺎر ﺑﺮده ﺷﺪ ﺗﺎ وارﯾﺎﻧﺖﻫﺎي ﻣﺴﺒﺐ ﺑﯿﻤﺎري در ﺑﯿﻤﺎراﻧﻤﺎن را ﺷﻨﺎﺳﺎﯾﯽ ﮐﻨﺪ. روش ﮐﺎر: ﭘﺲ از ﮐﺴﺐ رﺿﺎﯾﺖﻧﺎﻣﻪ از ﺑﯿﻤﺎران،DNA ژﻧﻮﻣﯽ از ﺧﻮن ورﯾﺪي اﺳﺘﺨﺮاج ﺷﺪ و ﺳـﭙﺲ ﺑﺮاي ﺷﻨﺎﺳﺎﯾﯽ ﺟﻬﺶﻫﺎي ﻣﺮﺗﺒﻂ ﺑﺎ ﺻﺮع، اﺑﺘﺪا، ﻧﻮاﺣﯽ رﻣﺰﮐﻨﻨﺪه و ﻣﺮزﻫﺎي اﮔﺰون- اﯾﻨﺘﺮون از 72 ژن ﻣﺮﺗﺒﻂ ﺑﺎ ﺻﺮع ﺑﺎ ﮐﺎرﺑﺮد ﮐﯿﺖ V4 ﺳﯿﺴـﺘﻢ ﻏﻨـﯽ ﺳـﺎزي ﻫﺪﻓﻤﻨـﺪSure selec‎t ﺑـﻪ دام اﻧﺪاﺧﺘـﻪ ﺷـﺪﻧﺪ . ﮐﺘﺎﺑﺨﺎﻧﻪﻫﺎي ﮐﭙﭽﺮ ﺷﺪه روي ﺳﯿﺴﺘﻤﯽ اﯾﻠﻮﻣﻨﺎ HiSeq 4000 ،ﺗﻮاﻟﯽﯾﺎﺑﯽ ﺷﺪﻧﺪ، ﺧﻮاﻧﺶﻫﺎي ﺗﻮاﻟﯽﯾـﺎﺑﯽ ﺷﺪه ﺑﺎ ﯾﮏ ژﻧﻮم ﻣﺮﺟﻊ ﻫﻤﺘﺮاز ﺷﺪﻧﺪ. اﺑﺰارPicard ﺑﻪ ﮐﺎر رﻓﺖ ﺗﺎ ﺧﻮاﻧﺶﻫﺎي ﻣﻀﺎﻏﻒ را ﺣﺬف ﮐﻨﺪ و ﻓﺮاﺧﻮاﻧﯽ وارﯾﺎﻧﺖﻫﺎ ﺑﺎ ﮐﺎرﺑﺮد اﺑﺰار آﻧﺎﻟﯿﺰ ژﻧﻮم )GTAK( اﻧﺠﺎم ﺷﺪ. ﺑﺮﻧﺎﻣﻪ ﺗﻔﺴﯿﺮ2 ﺑﻪ ﮐﺎر رﻓﺘﻪ ﺷﺪ ﺗﺎ وارﯾﺎﻧﺖﻫﺎ ﺗﻔﺴﯿﺮ ﮐﻨﺪ، ﺳﭙﺲ وارﯾﺎﻧﺖﻫﺎﯾﯽ ﺑﺎ ﻓﺮاوﻧﯽ آﻟـﻞ ﮐﻤﺘـﺮ از1 % ﻣﻄـﺎﺑﻖ ﺑـﺎ ﭘﺎﯾﮕﺎﻫـﺎي داده ﻫـﺎي ﻧﻮﮐﻠﺌﻮﺗﯿﺪ ﻣﺎﻧﻨﺪ dbSNP و ﻫﺰاره ژﻧﻮم، ﺟﺪاﺳﺎزي ﺷﺪﻧﺪ. اﺑﺰار In silico ﺑﻪ ﮐﺎر رﻓﺘﻪ ﺷـﺪ ﺗـﺎ ﺑﯿﻤـﺎرﯾﺰاﯾﯽ وارﯾﺎﻧﺖﻫﺎ را ارزﯾﺎﺑﯽ ﮐﻨﺪ. ﻧﺘﺎﯾﺞ: ﻣﻄﺎﺑﻖ ﺑﺎ ﭘﺎﯾﮕﺎهﻫﺎي داده ﭘﯿﺸﮕﻮﯾﯽ ﮐﻨﻨﺪه ﭘﺎﺗﻮژﻧﺴﯿﺘﯽ ، ﻣﻮﺗﺎﺳﯿﻮن ﺧﺎﺻـﯽ ﯾـﺎ وارﯾﺎﻧـﺖ از ﻧـﻮ در ژنﻫﺎي ﻣﺮﺗﺒﻂ ﺑﺎ ﺻﺮع ﯾﺎﻓﺖ ﻧﺸﺪ، وﻟﯽ ﭼﻨﺪﯾﻦ ﭘﻠﯽ ﻣﻮرﻓﯿﺴﻢ ﮔﺰارش ﺷﺪﻧﺪ. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: ﺑﺎ وﺟﻮد اﯾﻨﮑﻪ ﺑﺮﺧﯽ وارﯾﺎﻧﺖﻫﺎي ﯾﺎ ﭘﻠﯽ ﻣﻮرﻓﯿﺴﻢ ﻫﺎﯾﯽ در ژنﻫﺎﯾﯽ ي ﻣﺮﺗﺒﻂ ﺑﺎ ﺻﺮع در ﺑﯿﻤﺎراﻧﻤﺎن ﯾﺎﻓﺖ ﺷﺪﻧﺪ. اﻣﺎ ﻧﺘﻮاﻧﺴﺖ ﻣﺎ را در ﺗﺸﺨﯿﺺ ﻗﺎﺑﻞ ﻗﺒﻮل ﺑﺮاي اﯾﻦ ﺷﺮاﯾﻂ ﯾﺎري دﻫﺪ. ﺗﺤﻘﯿﻘﺎت ﺑﯿﺸﺘﺮي ﺟﻬﺖ آﺷﮑﺎرﺳﺎزي ﻋﻠﺖ ﺑﯿﻤﺎري اداﻣﻪ دارد.

The present study reports a case of familial episodic coma in which three girls manifesting refractory seizures followed by coma. Targeted gene panel of epilepsy using next-generation sequencing (NGS) technique was requested to identified disease -causing variant(s) in the patients. Materials and Method: After obtaining a written informed consent from our patient, genomic DNA was extracted from venous blood, for identifying mutations in epilepsy genes, at first, coding regions as well as all intron– exon boundaries of the 72 genes were captured by Sure selec‎t Target Enrichment System V4 kit (Agilent, Santa Clara, CA), then captured libraries were sequenced on an Illumina HiSeq 4000(Illumina Inc., San Diego, CA, USA), sequenced reads were aligned with a reference human genome and Picard tools was used to remove duplicated reads; variant calling was performed using the Genome Analysis Tool Kit (GATK). ANNOVAR was used to annotated variants, then all variants were filtered out based on minor allele frequency (MAF) <1 % according to data bases of nucleotide including dpSNP, 1000 Genome project. In silico tools was performed to evaluated pathogenicity of variant(s). Result: According to databases of pathogenicity prediction of gene, no specific mutation in epilepsy genes was found in our patients, but several polymorphisms were reported. Conclusion: Given polymorphisms in genes related to epilepsy were found in our study, failed to provide us with an acceptable diagnosis of this condition, further research is needed to reveal the cause of the disease.