Quantitative and qualitative rapid immune cell activation profiling to allow identification and follow up of at-risk neonates admitted to ICU

先兆子痫是一种由胎盘驱动的疾病，是围产期发病率和死亡率的主要原因，每年导致75,000名产妇和500,000名新生儿死亡。这种疾病会导致高血压、内皮细胞破坏和广泛的免疫功能障碍。然而，目前还没有标准的方法来确定谁会患先兆子痫，或者病情会有多严重。医学研究理事会iCASE(科学与工程产业合作奖)博士生项目与Sysmex合作，希望通过分析子痫前期及其临床后遗症的病理生理来解决这一问题。合体滋养层是胎盘与子宫直接接触的一层，在正常妊娠时释放细胞外小泡(STBEV)。STBEV在PE中含量更丰富，已被证明在子痫前期的发病机制中有重要作用，例如在凝血和免疫功能方面。已经确定了几个生物标志物，它们可以将STBEV与正常和先兆子痫胎盘区分开来。Sysmex Exoccount已被验证可用于定量和检测血浆样本中具有疾病特征的STBEV。此外，没有定义患有先兆子痫或其他不良妊娠结局的人的免疫细胞激活状态。目前使用白细胞计数和C反应蛋白的诊断工具不是最理想的，因为怀孕，特别是分娩过程是高度炎症的，因此在没有不良妊娠结局的情况下，这些参数可能会升高。妇女在整个怀孕期间都要接受常规的临床血液测试，并使用Sysmex XN分析仪进行常规分析。只有几个参数(例如，白细胞计数和C反应蛋白)被报告给临床医生，尽管分析仪还测量了其他几个不经常报告的参数。本项目的目标如下：1.使用Sysmex Exocounter对携带唯一标记(胎盘碱性磷酸酶)的循环STBEV(正常和先兆子痫/不良结局)进行量化。描述正常妇女(那些没有临床或血液学怀疑先兆子痫或其他不良妊娠结局的妇女)以及确诊为先兆子痫/不良结局的妇女的免疫细胞激活状态。利用蛋白质组学、转录学和高级成像数据，探讨STBEV(正常和先兆子痫/不良结局)对血液学、免疫和内皮细胞系的影响。整合AIMS 1-3的数据，以确定与先兆子痫和其他不良妊娠结局有关的潜在途径，以便为AIM 2.5中所见的免疫细胞激活状态的任何变化提供生物学/临床证据。利用NDWRH和Sysmex的联合国际网络建立全球患者验证队列，以确保Aim 2的结果可在全球范围内翻译。

Preeclampsia, a placentally-driven disease, is a leading cause of morbidity and mortality in the perinatal space and is responsible for 75,000 maternal and 500,000 neonatal deaths every year. The disease causes hypertension, endothelial disruption and widespread immune dysfunction. However, there is currently no standardized method for determining who will develop preeclampsia, or how severe it will be. This Medical Research Council iCASE (industrial collaborative award in science and engineering) doctoral studentship, in partnership with Sysmex, hopes to solve this issue through analysis of the pathophysiology of preeclampsia and its clinical sequelae. The syncytiotrophoblast is the layer of the placenta in direct contact with the uterus and releases extracellular vesicles (STBEVs) in normal pregnancy. STBEVs are much more abundant in PE and have been shown to contribute significantly to the pathogenesis of preeclampsia, for example in clotting and immune function. Several biomarkers have already been identified which allow for the distinction of STBEVs from normal and preeclamptic placentas. The Sysmex Exocounter has been validated for quantification and detection of STBEVs with disease characteristics in plasma samples. Furthermore, there is no definition of the immune cell activation status for individuals with preeclampsia or other adverse pregnancy outcomes. Current diagnostic tools utilizing white cells counts and CRP are suboptimal because pregnancy, especially the process of labour, is highly inflammatory and thus such parameters may be elevated in the absence of adverse pregnancy outcomes. Women undergo routine clinical blood tests throughout pregnancy, which are routinely analysed at using the Sysmex XN analyser. Only a few parameters (e.g., white cell count and CRP) are reported to clinicians, although the analysers also measure several other parameters, which are not routinely reported.The aims of this project are as follows:1. Quantify circulating STBEVs (normal and preeclampsia/adverse outcomes) carrying a unique marker (placental alkaline-phosphatase) using the Sysmex Exocounter.2. Characterize the immune cell activation status in normal women (those with no clinical or haematological suspicion of preeclampsia or other adverse pregnancy outcomes), and in women with confirmed preeclampsia/adverse outcomes.3. Explore the effects of STBEVs (normal and preeclampsia/adverse outcomes) on haematological, immune and endothelial cell lines using proteomic, transcriptomic and advanced imaging data.4. Integrate the data from Aims 1-3 to identify potential pathways implicated in preeclampsia and other adverse pregnancy outcomes in order to create biological/clinical evidence for any changes in immune cell activation status seen in Aim 2.5. Utilise the combined international networks of NDWRH and Sysmex to establish a worldwide patient validation cohort to ensure the findings of Aim 2 are globally translatable.