Identifying Genetic Predictors of Stroke Using Next Generation Sequencing

镰状细胞病的一个威胁生命和最严重的并发症是脑血管疾病。 一些患者可能会发生轻微的缺血性事件，范围从无症状到轻度症状。 其他患者可能在年轻时经历大脑动脉的完全闭塞。这些患者 可迅速发展为莫亚莫亚和脑灌注不足。如果没有长期输血治疗或 骨髓移植，患有严重脑血管疾病的患者有很高的风险， 到生命的第三个十年的死亡率。识别中风的危险因素将允许早期 在脑动脉闭塞之前进行预防性干预。 .在过去的几年里，已经有几个开创性的出版物的全基因组和全外显子组 测序应用患者-父母三联体来确定新生儿疾病的遗传基础。我们将采用 类似的方法来定义潜在的遗传修饰剂，预测风险，并提供更好的理解， 镰状细胞病患儿的早发性脑大动脉疾病我们已经确定， 收集了8个家庭的DNA样本，这些家庭有一个以上的孩子患有镰状细胞病，但只有 1例大面积脑动脉闭塞患儿。而不是病人父母三元组，我们将使用兄弟姐妹作为 额外的控制。从理论上讲，没有镰状细胞病的父母有可能携带突变或 单核苷酸多态性可预测镰状细胞相关脑血管疾病但在 没有镰状细胞表型，就没有中风表型。 该试点的另一个目的是发展生物信息学和计算生物学专业知识， 涉及下一代测序应用的其他项目。使这项技术 可用于其他项目，我们将开发和验证质量保证驱动的生物信息学 渠道.这将是该试点项目下半年的重点。我们将使用在 镰状细胞病患者开发和验证管道。这条管道和方法将是 可扩展以用于更大规模的未来研究。

One of the life-threatening and most severe complications of sickle cell disease is cerebral vascular disease. Some patients may experience small ischemic events that range from asymptomatic to mildly symptomatic. Other patients may experience complete occlusion of a large cerebral artery at a young age. These patient may quickly develop moya moya and deficient cerebral perfusion. Without chronic transfusion therapy or a bone marrow transplant, patients with significant cerebral vascular disease have a high risk for diseaserelated mortality by the third decade of life. Identification of risk factors for stroke will permit early preventative interventions prior to cerebral artery occlusion. . In the past few years, there have been several seminal publications of whole genome and whole exome sequencing applied patient-parent triads to define the genetic basis of disease in newborns. We will apply a similar approach to define potential genetic modifiers that predict risk and offer a better understanding of early onset large cerebral artery disease in children with sickle cell disease. We have identified and collected DNA samples on eight families with more than one child affected by sickle cell disease, but only one child with large cerebral artery occlusion. Instead of patient-parent triads, we will use the sibling as an additional control. Theoretically, it is possible for a parent without sickle cell disease to carry a mutation or single nucleotide polymorphism that predicts sickle cell associated cerebral vascular disease. However, in the absence of the sickle cell phenotype, there is no stroke phenotype. Another aim for this pilot is to develop the bioinformatic and computational biology expertise to pursure additional projects involving the application of next generation sequencing. To make this technology available for additional projects, we will develop and validate a quality assurance driven bioinformatics pipeline. This will be the focus of the second half of this pilot project. We will use the data obtained in patients with sickle cell disease to develop and validate the pipeline. This pipeline and approach will be scalable for larger future studies.