Genetic screens for erythrocyte determinants of protein trafficking in malaria parasites

项目总结 人类感染恶性疟原虫会导致严重的发病率和死亡率。这个 疟疾的发病机制与寄生虫感染的红细胞通过其 细胞与血管壁的粘连。PfEMP1已被确定为红血球上的关键寄生虫分子 介导这种结合的细胞表面，并且已经在鉴定特定的PfEMP1方面做了很多工作 蛋白质基序和关键的寄生虫分子是蛋白质运输和出口到红色所必需的 血细胞。 在这项工作中，我们试图识别影响蛋白质的红细胞蛋白质组中的红细胞分子。 贩运到红血球表面。我们将开发基因筛查系统，以进行 CD34+造血细胞的互补基因敲除和基因敲除筛选及体外培养 以产生突变的红细胞。具体地说，我们将识别出 将毒力蛋白PfEMP1运输到红细胞表面。 PfEMP1转运到宿主红细胞中特定宿主红细胞分子和途径的鉴定 血细胞将极大地提高我们对宿主-寄生虫在转化过程中相互作用的理解 并将协助设计以治疗为目标的蛋白质贩运策略。此外， 红细胞基因筛查方法的发展将使对许多 红细胞与疟原虫之间的相互作用。寄生虫。

PROJECT SUMMARY Infection of humans with Plasmodium falciparum parasites results in significant morbidity and mortality. The pathogenesis of malaria is associated with the sequestration of parasite-infected red blood cells through their cytoadherence to vascular walls. PfEMP1 has been identified as the key parasite molecule on the red blood cell surface that mediates this binding, and much effort has gone into the identification of the specific PfEMP1 protein motifs and critical parasite molecules that are required for protein trafficking and export into the red blood cell. In this work, we seek to identify red blood cell molecules within the red blood cell proteome that effect protein trafficking to the surface of red blood cells. We will develop genetic screening systems to conduct complementary knockdown and knockout screens using CD34+ hematopoietic cells followed by in vitro culture of red blood cells to produce mutant red blood cells. Specifically, we will identify molecules that are required for trafficking of the virulence protein PfEMP1 to the surface of the red blood cell. The identification of specific host red blood cell molecules and pathways in PfEMP1 trafficking to the host red blood cell will greatly enhance our understanding of host-parasite interactions during the transformation of the host red blood cell, and will aid in devising strategies that therapeutically target protein trafficking. Moreover, the development of red blood cell genetic screening methodologies will allow the study of numerous interactions between the red blood cell and Plasmodium spp. parasites.