Development of a chimeric mouse model for the analysis of Plasmodium liver stages

描述（由申请人提供）：本提案的目的是验证和表征人类疟原虫感染的红细胞外期嵌合人-鼠肝脏模型。此外，将从嵌合肝脏中回收肝脏期寄生虫，通过激光捕获显微解剖，分离其mRNA并用于微阵列分析。在恶性疟原虫的不同发育阶段中，最不明确的是肝期或红细胞外期（EE）。这一阶段包括寄生虫进入肝细胞后的感染和增殖，只能在人类和少数非人类灵长类动物中进行研究。不幸的是，很难获得含有恶性疟原虫或间日疟原虫EE形式的生物材料，这两种寄生虫是造成大多数人类病例的原因。本项目提出的研究将产生关于先前鉴定抗原的动态表达的体内数据，并通过微阵列分析允许鉴定寄生虫在EE发育过程中的转录组。这种类型的全球分析已经成为可能，因为人类疟疾寄生虫恶性疟原虫的整个基因组序列已经完成。尽管已经确定了寄生虫基因组中的5,200多个基因，但潜在疫苗和药物靶点的确定将取决于结合关系数据库和信息学的功能基因组学研究，以确定每种编码蛋白的特征（身份）。EE阶段转录组的分析将补充最近几项全基因组研究，这些研究已经阐明了恶性疟原虫孢子体、分裂子体、滋养体和配子体的蛋白质组和转录组。这将产生人类疟疾寄生虫生命周期中基因表达的真正全面的图景。

DESCRIPTION (provided by applicant): The objective of this proposal is the validation and characterization of a chimeric human-murine liver model for the exo-erythrocytic stage of human Plasmodium infections. Additionally, liver stage parasites will be recovered from the chimeric livers, by laser capture micro-dissection, their mRNA isolated and used for microarray analysis. Among the different developmental stages of Plasmodium falciparum, the least well characterized is the hepatic or exo-erythrocytic (EE) phase. This stage encompasses the infection and multiplication of the parasite following entry into the hepatocyte and can only be studied in humans and a few non-human primates. Unfortunately, it is difficult to obtain biological material containing the EE forms of P. falciparum or P. vivax, the parasites responsible the majority of cases in humans. The studies proposed in this project will produce in vivo data on the kinetic expression of previously identified antigens and by using microarray analysis allow the identification of the parasite transcriptome during EE development. This type of global analysis has been made possible, because the entire genomic sequence of the human malaria parasite, P. falciparum has been completed. Despite the identification of the over 5,200 genes in the parasite genome, the identification of potential vaccines and drug targets will depend upon functional genomics studies combined with relational databases and informatics to determine the characteristics (identity) of each encoded protein. The analysis of the EE stage transcriptome will complement several recent genome-wide studies that have elucidated the P. falciparum proteome and transcriptome of sporozoites, merozoites, trophozoites and gametocytes. This will then produce a truly comprehensive picture of gene expression during the lifecycle of human malaria parasites.