Identification and Assessment of Surrogate Biomarkers for Chronic Chagas Disease

虽然低级别寄生虫的持久性是慢性恰加斯病的一个基本方面，但目前的寄生虫学分析灵敏度低，而且不是定量的。苯并硝唑是唯一可用的锥虫药物，在治疗慢性恰加斯病患者方面的疗效值得怀疑。目前迫切需要对治疗慢性恰加斯病的新药进行临床试验，然而缺乏可靠的生物标志物来减少寄生和由此产生的炎症反应和损伤是评估新药的主要障碍。鉴别标记物的存在和水平的鉴定。crtvz;寄生和由此产生的免疫和炎症扰动可能潜在地解决这个问题。少数可用的寄生虫病和疾病进展的生物标志物既不是定量的，也不是高度预测的。众所周知，复合生物标志物在区分临床结果方面比单一标志物具有更高的潜力。血液基因表达谱已被用于区分几种传染病中具有不同感染病程的患者，包括我们小组对恰加斯病的初步数据。我们假设持续寄生在7。可诱导PBMC基因表达模式的特异性改变。第二种假设是根据临床表现和发现来确定感染的不同阶段。克氏PCR和抗体结果，将显示特定的复合生物标志物谱。这个项目的主要目的是确定高和低7的转录谱。克鲁兹寄生以及具有不同临床表现的患者的转录谱，并将转录组分析的结果与我们目前正在表征的其他生物标志物参数相结合。该研究将评估代表恰加斯病不同临床阶段和寄生强度的患者样本(按7。cruzi real-time PCR)，以及来自REDS II Chagas队列的血清阴性对照。将对全血进行转录组分析，与每个临床组相关的转录特征将通过qPCR分析进行验证，该分析可应用于更大的样本集。然后，我们将在其余的REDS II恰加斯病队列（600名患者和100名血清阴性对照）中测试验证的mRNA标记物的分类能力。血浆蛋白生物标志物炎症和心血管疾病；克鲁兹/抗体谱/滴度，在相同的患者组中进行表征，将与转录组特征相结合，构建一个改进的复合生物标志物谱，用于预后和治疗监测。

Although low-grade parasite persistence is a fundamental aspect of chronic Chagas disease, current parasitological assays have low sensitivity and are not quantitative. Benznidazole, the only available trypanocidal drug, has questionable efficacy in the treatment of chronic Chagas disease patients. There is an urgent need to perform clinical trials of new drugs for chronic Chagas disease, however the lack of reliable biomarkers for reduction of parasitism and consequent inflammatory responses and damage is a major obstacle for evaluating new drugs. The identification of differentiating markers for presence and levels of 7. crtvz; parasitism and resulting immune and inflammatory perturbations could potentially solve this problem. The few available biomarkers for parasitism and disease progression are neither quantitative nor highly predictive. It is known that composite biomarkers have a higher potential for differentiating clinical outcomes than single markers. Blood gene expression profiling has been used to differentiate between patients with distinct courses of infection in several infectious diseases including preliminary data from our group for Chagas disease. We hypothesize that persistent parasitism by 7. cruzi, induces specific changes in PBMC gene expression patterns. A second hypothesis is that distinct stages of infection, defined based on clinical presentation and findings and 7. cruzi PCR and antibody results, will display specific composite biomarker profiles. The major aims of this project are to identify the transcriptional profiles of high and low 7. cruzi parasitism as well as the transcriptional profiles of patients with different clinical presentations, and to combine the findings from transcriptome analyses with other biomarker parameters we are currently characterizing. The study will evaluate a sample of patients representative of distinct clinical stages of Chagas disease and intensity of parasitism (as measured by 7. cruzi real-time PCR), along with seronegative controls, from the REDS II Chagas cohort. Transcriptome analysis of whole blood will be performed, and transcriptional signatures that correlate with each clinical group will be validated with qPCR assays that can be applied to larger sample sets. We will then test the classifying ability of validated mRNA markers on the remainder of the REDS II Chagas disease cohort (600 patients and 100 seronegative controls). Plasma protein biomarkers of inflammation and cardiovascular disease and 7. cruz/antibody profiles/titers, which are being characterized in the same patient groups, will be combined with transcriptome signatures to construct an improved composite biomarker profile for prognosis and therapeutic monitoring.