TRANSCRIPTOME PROFILES OF GENE EXPRESSION IN MONKEY MODELS OF HUMAN MALARIA

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。疟疾是一种蚊子传播的疾病，是一个重大的全球健康问题，NIAID制定了一项议程，将扩大对这一重要病原体的研究工作，包括基因组研究。间日疟原虫是四种人类疟疾寄生虫中最常见的，感染在亚洲、拉丁美洲、中东、北非和南太平洋广泛存在。本研究中使用的cynomolgi是一种与间日疟原虫密切相关的类人猿寄生虫，也是间日疟原虫的一个很好的模型。用食蟹假单胞虫孢子子接种两只恒河猴，并从感染前（基线）、8天后（肝期）、寄生虫病高峰期、第一次复发和第二次复发时采集的外周血单个核细胞（PBMCs）中分离RNA。样本在包含超过22,000个基因的人体微阵列芯片上运行。使用超过3000个差异表达基因的样本聚类表明，来自两只猴子的样本聚类相似，基线样本与其他样本非常不同，第一次和第二次复发样本最相似。基因的层次聚类显示，两种猴子的基因表达水平发生了显著变化，特别是在肝脏初始阶段和寄生虫病高峰期，许多基因下调。有趣的是，随着时间的推移，与防御和免疫有关的富集基因的数量有所增加。在这项研究中，我们能够有效地使用猴疟疾模型来研究宿主基因表达的转录组谱，因为我们感染了疟疾感染的动物，并且与疟疾流行地区的人类患者不同，伴随感染和营养状况都不是混杂因素。这项工作的发表引起了疟疾研究界的极大兴趣，并在12月被邀请向基因组研究所作报告。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Malaria, a mosquito borne disease, is a major global health concern and NIAID has developed an agenda that will expand research efforts, including genomic studies, on this important pathogen. P. vivax is the most common of the four human malaria parasites and infection is widespread in Asia, Latin America, the Middle East, North Africa, and the South Pacific. P. cynomolgi, used in this study, is a simian parasite closely related to, and a good model of, P. vivax. Two rhesus macaques were inoculated with sporozoites of P. cynomolgi and RNA was isolated from peripheral blood mononuclear cells (PBMCs) harvested before infection (baseline), after 8 days (liver phase), at the peak parasitemia, at the first relapse, and at the second relapse. Samples were run on a human microarray chip containing over 22,000 genes. Sample clustering, using over 3,000 differentially expressed genes, showed that samples from both monkeys clustered similarly, with baseline samples being very distinct from other samples and the first and second relapse samples being the most similar. Hierarchical clustering of the genes showed dramatic gene expression level changes on both monkeys, especially down regulation of many genes during the initial liver stages and at the peak parasitemia. Interestingly, the number of enriched genes involved in defense and immunity showed increase throughout the time course. In this study we were able to use the monkey malaria model effectively to study the transcriptome profiles of the host gene expression, since we infected malaria-na¿ve animals, and neither concomitant infections nor nutritional status were confounding factors as would be expected in human patients from malaria-endemic areas. The publication of this work has elicited considerable interest from the malaria research community and was the subject of an invited presentation to The Institute of Genomic Research in December.