Genetic screens for erythrocyte determinants of protein trafficking in malaria parasites

项目摘要 人类感染恶性疟原虫寄生虫会导致显着的发病率和死亡率。的 疟疾的发病机制与寄生虫感染的红细胞通过其 细胞粘附于血管壁。PfEMP 1已被确定为红血上的关键寄生虫分子 细胞表面介导这种结合，并且已经进行了大量的努力来鉴定特异性PfEMP 1 蛋白质基序和关键的寄生虫分子，这些分子是蛋白质运输和出口到红色 血细胞 在这项工作中，我们试图确定红细胞蛋白质组中影响蛋白质的红细胞分子， 运输到红细胞表面。我们将开发基因筛查系统， 使用CD 34+造血细胞随后进行体外培养的互补敲低和敲除筛选 产生变异红细胞。具体来说，我们将识别出 毒力蛋白PfEMP 1向红细胞表面的运输。 PfEMP 1转运至宿主红细胞的特异性宿主红细胞分子和途径的鉴定 血细胞将大大提高我们对宿主-寄生虫相互作用的理解， 宿主红细胞，并将有助于设计治疗目标蛋白质运输的策略。此外，委员会认为， 红细胞遗传筛查方法的发展将允许研究许多 红细胞和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.