| Background After intracranial aneurysm(IA) ruptured, case fatality rate is about 40%. Domestic studies about epidemiology of intracranial unruptured aneurysms found that the aneurysm prevalence is about 7%, higher than the prevalence of other countries in Europe and America. Etiology has not yet clear. At present, many believe that IA is an acquired complex disease under the action with a variety of genetic factors and environmental factors.Objective This study is to explore the susceptibility gene mutation and population characteristics of intracranial aneurysms and provide a more accurate and comprehensive genomic information in genetic research about IA, while clinical molecular markers for the provision of diagnostic and screening IA. Our work could find out the new targets for prevention and treatment of IA, to lay the foundation for improving the diagnosis and treatment of IA.Content(1) To determine the object of study and collect samples, then taking advantage of whole-genome sequencing technology to sequence.(2) Based on whole genome sequencing, suspicious susceptibility genes were filtered by bioinformatic analysis. Our study will use WGS and WES in familial intracranial aneurysms to find SNP, SV, in Del, CNV and other variation information. Through bioinformatics and analysis of comments, found out the initial screening susceptibility genes associated with IA.(3) With molecular biology techniques, Sanger sequencing technology tests and verifies related genetic variation detection susceptibility genes in sporadic aneurysms expression. For a variety of genetic variation SNP, SV, CNV, In Del etc. through whole genome sequencing found in 290 sporadic intracranial aneurysms exceptions peripheral blood DNA samples and peripheral blood samples from 257 cases of healthy controls, using different methods for different variation type verification(4) To study the role of genetic variation associated in vitro experiments IA in theangiogenesis.Method(1) Obtaining DNA samples from FIA patients and sporadic aneurysm, as well as samples of the control group patients. FIAs were found through the questionnaire, signed informed consent, collecting FIA and family members blood samples, to extract DNA. In clinical sporadic patients after informed consent, patient specimens from blood samples, to extract DNA.(2) WGS sequencing of the FIA patients and data analysis. 20 cases of FIA were subjected to WES sample and WGS sequencing. The obtained data was analysised through bioinformatic analysis.(3) To validate WGS and WEW sequencing results obtained from patients with sporadic aneurysms. WGS and WES results are directly verified.(4) Gene interference in tube formation in vitro experiments to verify results. Using gene silencing technology to reduce screened IA related susceptibility gene expressed in vascular endothelium. vitro cell experiments at the cellular level to observe the process of angiogenesis, to observe IA susceptibility genes in the human angiogenesis mechanisms action, such as endothelial cell proliferation, migration, capillary reticular formation.Result(1) The study eventually included a total of 10 familial intracranial aneurysms pedigrees included 20 patients who were subjected to WGS and WES sequence. And found that ARHGEF17 is IA related new susceptibility gene.(2) To verify that the new susceptibility gene mutation 5 sites on ARHGEF17 in the peripheral blood DNA sample of sporadic intracranial aneurysms patients. Peripheral blood samples from 290 and 257 healthy people from control group showed the statistically significant(p < 0.05).(3) Experimental results show that cell susceptibility genes ARHGEF17 can promote angiogenesis and endothelial cell proliferation.Conclusion ARHGEF 17 genes play an important role in the formation of intracranial aneurysm, intracranial aneurysm new susceptibility gene. |
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