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

描述(由申请人提供)：这项建议的目标是验证和表征人-鼠嵌合肝脏模型，用于人类疟原虫感染的红外期。此外，通过激光捕获显微解剖，将从嵌合肝脏中提取肝期寄生虫，分离它们的mRNA并用于微阵列分析。在恶性疟原虫不同发育阶段中，肝期或红外期(EE)特征最差。这一阶段包括寄生虫进入肝细胞后的感染和繁殖，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，这些寄生虫导致了人类大多数病例。该项目中提出的研究将产生关于先前识别的抗原的动态表达的体内数据，并通过使用微阵列分析来鉴定EE发育过程中的寄生虫转录组。这种类型的全球分析之所以成为可能，是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标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，以确定每个编码蛋白质的特征(身份)。对EE阶段转录组的分析将是对最近几项全基因组研究的补充，这些研究已经阐明了恶性疟原虫的蛋白质组和子孢子、裂殖子、滋养体和配子体的转录组。这将产生一个真正全面的人类疟疾寄生虫生命周期中基因表达的图景。

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.