Defining the Inflammation and Immunity Transcriptome in Severe Malarial Anemia for Immunotherapeutic Discovery

项目摘要 尽管全球卫生取得了进步，但疟疾每年感染2.12亿人，造成42.9万人死亡。缺乏有效 疫苗和抗药性菌株的出现使得新的治疗方法至关重要。童年 在恶性疟原虫完全流行的情况下，36个月以下儿童的死亡率仍然非常高 传播地区，如肯尼亚的锡亚亚，由于严重的疟疾贫血[SMA，血红蛋白（Hb）<5.0 g/dL]。 在过去的14年里，在我们位于锡亚亚的最先进的临床研究设施中，我们专注于研究先天性 免疫力和共病影响儿童疟疾的临床结果。三年多的随访 在此期间，我们发现93%的儿童患有疟疾，22.7%的儿童发展为SMA。为了找出最 区分SMA和较轻疾病的重要途径，我们在过去几年中使用了 结合全球基因组学方法，创建用于识别疾病的收敛模型 签名.这些研究表明，免疫应答基因是SMA的最强预测因子， 因此，将搜索范围从整个基因组缩小到参与宿主免疫反应的途径。基于 根据这些发现和最近的初步数据，我们建议使用靶向RNAseq和一个平台， 包含500个免疫应答基因（先天免疫和适应性免疫），这些基因尚未伴随 在疟疾中探索。由于我们的研究结果表明，对恶性疟原虫的免疫反应驱动了不利的 即使在寄生虫清除后，几周的临床结果，疟疾性贫血是一个理想的候选人 用于免疫治疗。因此，该项目的总体目标是确定关键的基因通路内的 转录组介导疾病的严重程度，然后用化合物靶向这些特定基因， 在患有轻度疾病的儿童中观察到的表达谱。为了实现这一目标，我们将 以下具体目标：1）确定疟疾中基因通路的时间动态变化如何 免疫转录组在急性疾病期间促进SMA; 2）确定基因通路如何改变， 疟疾免疫转录组介导疟疾的严重性， 获得性免疫;和3）鉴定疟疾免疫转录组中的免疫靶点， 用于降低疟疾疾病的严重程度，并在未来的试验中改善临床结果。成功 完成这些目标，我们将确定宿主和寄生虫的表达谱如何影响急性疾病 14天以上（队列1），并在儿童第1天的纵向出生队列（队列2）中验证这些发现。 急性疟疾发作。我们还将确定改变宿主和寄生虫的时间表达的影响 在36个月内自然获得性免疫力的发展概况（群组2）。离体样品来自 这两组将用于测试免疫调节化合物对宿主表达谱的影响。 实现这些目标将对以下方面产生广泛的影响：（1）确定高危群体， 以及（2）优先考虑可用于改善未来免疫治疗试验中的临床结果的化合物。

PROJECT SUMMARY Despite advances in global health, malaria infects 212M and kills 429,000 people annually. Lack of effective vaccines and emergence of drug-resistant strains makes new therapeutic approaches essential. Childhood mortality rates in children <36 months remain exceedingly high in holoendemic Plasmodium falciparum transmission regions such as Siaya, Kenya, due to severe malarial anemia [SMA, hemoglobin (Hb)<5.0 g/dL]. Over the last 14 years, at our state-of-the-art clinical research facilities in Siaya, we focused on how innate immunity and co-morbidities influence clinical outcomes in childhood malaria. Over a three-year follow-up period, we found that 93% of the children had malaria, and 22.7% developed SMA. To identify the most significant pathways that discriminate SMA from milder forms of disease, we spent the last several years using a combination of global genomic approaches to create a convergent model for identification of disease signatures. These studies demonstrated that immune response genes are the strongest predictors for SMA, thus, narrowing the search from the entire genome to pathways involved in the host-immune response. Based on these findings and recent preliminary data, we propose to use targeted RNAseq with a platform that contains of 500 immune response genes (innate and adaptive immunity) which have not been concomitantly explored in malaria. Since our findings demonstrate that the immune response to P. falciparum drives adverse clinical outcomes for several weeks, even after clearance of the parasite, malarial anemia is an ideal candidate for immunotherapy. Thus, the overall goal of the project is to identify critical gene pathways within the transcriptome that mediate disease severity and then target these specific genes with compounds that elicit expression profiles witnessed in children with milder forms of disease. To achieve this goal, we will complete the following specific aims: 1) Determine how changing temporal dynamics of gene pathways in the Malarial Immunity Transcriptome promote SMA during acute disease; 2) Determine how changes in gene pathways in the Malarial Immunity Transcriptome mediate malarial severity throughout the development of naturally- acquired immunity; and 3) Identify immunotherapeutic targets in the Malarial Immunity Transcriptome that can be used to reduce malaria disease severity and improve clinical outcomes in future trials. To successfully complete these aims, we will determine how host and parasitic expression profiles impact on acute disease over 14 days (cohort 1) and validate these findings in a longitudinal birth cohort (cohort 2) during the child's 1st episode of acute malaria. We will also determine the impact of changing host and parasite temporal expression profiles on the development of naturally-acquired immunity over 36 months (cohort 2). Ex vivo samples from the two cohorts will be used to test the effect of immunotherapeutic compounds on host expression profiles. Accomplishing these goals will have broad reaching translational implications for: (1) identifying at-risk groups, and (2) prioritizing compounds that can be used to improve clinical outcomes in future immunotherapy trials.