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，每年造成75，000名产妇和500，000名新生儿死亡。该疾病导致高血压、内皮破坏和广泛的免疫功能障碍。然而，目前还没有标准化的方法来确定谁会发展先兆子痫，或者它会有多严重。这个医学研究理事会iCASE（科学与工程工业合作奖）博士生项目与Sysmex合作，希望通过分析先兆子痫的病理生理学及其临床后遗症来解决这个问题。合体滋养层是胎盘与子宫直接接触的层，在正常妊娠中释放细胞外囊泡（STBEV）。STBEV在PE中更丰富，并且已经显示出显著促进先兆子痫的发病机制，例如凝血和免疫功能。已经鉴定了几种生物标志物，其允许将STBEV与正常胎盘和先兆子痫胎盘区分开。Sysmex Exocounter已被验证用于定量和检测血浆样本中具有疾病特征的STBEV。此外，对于患有先兆子痫或其他不良妊娠结局的个体，没有定义免疫细胞活化状态。目前利用白色细胞计数和CRP的诊断工具是次优的，因为妊娠，特别是分娩过程是高度炎症性的，因此在没有不良妊娠结果的情况下这些参数可能升高。妇女在整个怀孕期间接受常规临床血液检查，使用Sysmex XN分析仪进行常规分析。只有几个参数（例如，白色细胞计数和C反应蛋白）报告给临床医生，尽管分析仪还测量了其他几个参数，这些参数不是常规报告的。使用Sysmex Exocounter定量携带独特标志物（胎盘碱性磷酸酶）的循环STBEV（正常和先兆子痫/不良结局）。描述正常女性（临床或血液学上没有先兆子痫或其他不良妊娠结局的可疑者）和确诊先兆子痫/不良结局的女性的免疫细胞活化状态。3.使用蛋白质组学、转录组学和先进的成像数据探索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。整合目标1-3的数据，以确定涉及先兆子痫和其他不良妊娠结局的潜在途径，从而为目标2.5中观察到的免疫细胞活化状态的任何变化创建生物学/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的发现在全球范围内可翻译。

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.