exRNA signatures Predict Outcomes after brain injury

描述（由申请人提供）： 创伤性脑损伤，无论是直接脑损伤还是缺血性或出血性梗死的后遗症，都与显著的发病率相关， 死亡率以及患者和医疗保健系统的成本。 出血性梗塞（如蛛网膜下腔出血和脑室内出血）是动脉破裂的罕见但毁灭性的并发症。 尽管付出了巨大努力，但蛛网膜下腔出血（aSAH）或脑室内出血（IVH）患者的结局几乎没有改善。鉴于这些脑损伤的可预测的时间过程，它们是生物标志物鉴定的理想候选者。一种检测“高危”患者的生物标志物和迟发性神经功能缺损的预后指标将大大改善目前的治疗方法，并增加我们对潜在病理事件的理解。 这些信息将立即导致新的治疗策略的实施。 过去几年的研究已经确定细胞外RNA（exRNA）作为生物标志物的潜在来源。 新技术，如下一代测序（NGS），使得有可能灵敏地和定量地分析少量的RNA包含在临床上可达到的体积的生物流体，如CSF和血浆。 在这项资助的前两年，我们建议对aSAH患者的CSF和血浆以及IVH早产儿的血浆和CSF中的exRNA（miRNA和总RNA）进行测序。每天收集aSAH样本~可用的样本量将使我们不仅能够对包括miRNA在内的总RNA进行测序，还将允许我们在本基金的第3-5年探索独特的富集策略~我们将检查CSF和血浆中与微泡相关的特定RNA。我们有这些患者的日常临床评估以及他们的结果（共25名受试者）。 我们将使用这些样本来鉴定预测血管痉挛发作和迟发性神经功能缺损严重程度的RNA标记物。我们还每隔一天从患有III-IV级IVH的早产儿中收集CSF和血浆样本，这些早产儿有储血器以排出血液， 脑脊液过多 我们也有婴儿的临床结果，无论他们是否 发生脑积水，需要永久性分流，以及18个月的发育迟缓或残疾评估。在第3-5年，我们将继续与我们的临床合作伙伴合作收集样本。届时，我们将从我们的发现平台Illumina HiSeq 2000切换到Illumina MiSeq进行验证。 一旦发现阶段完成并鉴定出潜在的生物标志物，就可以在MiSeq更快的流体学、成像和缩短的运行时间上进行验证，而无需在发现和验证阶段之间改变化学。

DESCRIPTION (provided by applicant): Traumatic brain injury, both from direct trauma to the brain and as a sequela of ischemic or hemorrhagic infarction are associated with a significant rate of morbidity, mortality and cost to both patients and the healthcare system. Hemorrhagic infarcts (such as aneurysmal subarachnoid hemorrhage and intraventricular hemorrhage) are rare but devastating complications of arterial rupture. Despite significant effort, there has been little improvement in the outcomes of patients with aneurysmal subarachnoid hemorrhage (aSAH) or intraventricular hemorrhage (IVH). Given the predictable temporal course of these brain injuries, they are ideal candidates for biomarker identification. A biomarker for detection o 'at risk' patients and a prognostic indicator of delayed neurological deficit would vastly improve current treatments and increase our understanding of underlying pathological events. This information would immediately lead to the implementation of new therapeutic strategies. Research over the last several years has identified extracellular RNAs (exRNA) as a potential source of biomarkers. New technology, such as next generation sequencing (NGS), has made it possible to sensitively and quantitatively assay small amounts of RNA contained in clinically attainable volumes of biofluids such as CSF and blood plasma. In the first two years of this grant, we propose to sequene the exRNA (miRNA and total RNA) from both the CSF and plasma of patients who present with aSAH and the plasma and CSF from premature infants born with IVH. aSAH samples were collected daily~ the volume of sample available will allow us not only to sequence the total RNA including miRNAs, but will permit us to explore unique enrichment strategies in years 3-5 of this grant~ we will examine specific RNAs associated with microvesicles in both CSF and plasma. We have both the daily clinical evaluations of these patients as well as their outcomes (25 subjects in all). We will use these samples to identiy RNA markers that predict onset of vasospasm and the severity of delayed neurological deficits. We also have CSF and plasma samples collected every other day from premature infants with grade III-IV IVH that had reservoirs put in place to evacuate blood and excess CSF. We also have the clinical outcomes from the infants, whether or not they developed hydrocephalus, the need for a permanent shunt, and an 18 month assessment of developmental delays or disabilities. In years 3-5, we will continue to collect samples in collaboration with both of our clinical partners. At that time, we will switch from our discovery platform, the Illumina HiSeq 2000 that has high depth of sequencing coverage, to the Illumina MiSeq for validation. Once the discovery phase is complete and potential biomarkers identified, validation can be carried out on the MiSeq's faster fluidics, imaging, and shortened run times without changing chemistries between the discovery and validation phases.