Developing comparative and functional genomic approaches to study Schistosoma

描述（由申请人提供）：血吸虫病是一种被忽视的热带疾病，影响发展中国家2亿多人。目前只有一种药物（吡喹酮）可用于治疗这种疾病，突出了开发新技术来研究这些寄生虫的重要性。虽然存在研究血吸虫的重要工具（例如基因组序列和RNA干扰），但血吸虫生命周期的复杂性继续阻碍开发新疗法的进展。具体而言，目前不可能进行大规模的、无偏见的功能基因组筛选来识别新的药物靶点。我们的长期目标是确定血吸虫病新的治疗靶点，目前的目标是使用实验上易于处理的自由生活的扁虫S。作为一个模型来识别和破译“扁虫特异性”基因的功能。尽管它们的生活方式不同（自由生活与寄生），但这些动物在解剖学和基因组水平上都有许多基本的相似之处。鉴于这些相似之处，我们的假设是，许多重要的生理过程，生殖和生存能力是深深保存在这些扁虫。为了解决这一假设，我们提出了以下两个具体目标：（1）确定生存能力或繁殖所需的扁虫特异性基因和（2）开发新的技术来破坏体内成虫染色体基因功能。第一个目的是利用高通量原位杂交技术比较108个扁虫特异性基因在曼氏血吸虫和日本血吸虫中的组织特异性表达。地中海。我们还将确定这些基因在沙门氏菌中的功能丧失表型。利用RNA干扰技术研究地中海贫血。这些研究将为具体目标#2中提出的体内功能研究提供候选基因列表。由于目前不可能在体内遗传操纵成体染色体，我们的第二个目标将确定体内吗啉代是否可以诱导感染小鼠体内成体染色体组织中的基因敲低。这些方法将用于检查在特定目标#1中鉴定的基因是否在体内染色体生物学中具有重要作用。这些研究是创新的，因为它们（i）引入S。medioprotea作为一个模型，以加快血吸虫的功能基因组研究和（ii）开发吗啉基技术，在哺乳动物宿主中的遗传操纵成人染色体。这些研究是重要的，因为它们描述了一个潜在的强有力的工作流程，为抗β-内酰胺酶的目标发现：大规模筛选的S。使用体内-吗啉代进行体内候选物验证。总的来说，我们的研究将试图填补寄生扁形虫功能基因组研究的重要空白，并有可能确定血吸虫新的治疗靶点。 公共卫生相关性：这些研究与公共卫生有关，因为它们旨在开发新的方法来了解人类病原体曼氏血吸虫基因的功能。完成拟议的研究可能有助于发现新的药物来对抗这种重要的人类寄生虫引起的疾病。

DESCRIPTION (provided by applicant): Schistosomiasis is a neglected tropical disease that affects more than 200 million people in the developing world. Currently only a single drug (praziquantel) is available to treat this disease, highlighting the importance of developing new techniques to study these parasites. Although important tools exist to study Schistosoma (e.g. genome sequences and RNA interference), the complexity of the schistosome life cycle continues to hinder progress towards developing novel therapeutics. Specifically, large-scale, unbiased functional genomic screens to identify new drug targets are currently not possible. With the long-term goal of identifying novel therapeutic targets in Schistosoma, our current objective is to use the experimentally tractable free-living flatworm S. mediterranea as a model to identify and decipher the function of "flatworm-specific" genes. Despite their divergent lifestyles (free-living vs. parasitic), these animals share many fundamental similarities, both at the anatomic and genomic levels. Given these similarities, our hypothesis is that many processes essential for physiology, reproduction, and viability are deeply conserved between these flatworms. To address this hypothesis we propose the following two specific aims: (1) Identify flatworm-specific genes required for viability or reproduction and (2) Develop novel techniques to disrupt adult schistosome gene function in vivo. In the first aim, we will use high-throughput in situ hybridization to compare the tissue-specific expression of 108 flatworm-specific genes in Schistosoma mansoni and S. mediterranea. We will also determine the loss-of-function phenotype for these genes in S. mediterranea using RNA interference. These studies will provide a candidate gene list for in vivo functional studies proposed in specific aim #2. Since it is presently not possible to genetically manipulate adult schistosomes in vivo, our second aim will determine if vivo-morpholinos can induce gene knockdown in adult schistosome tissues inside infected mice. These approaches will be used to examine if genes identified in specific aim #1 have important roles in schistosome biology in vivo. These studies are innovative because they (i) introduce S. mediterranea as a model to accelerate functional genomic studies of Schistosoma and (ii) develop morpholino-based techniques to genetically manipulate adult schistosomes in the mammalian host. These studies are significant since they describe a potentially potent workflow for anti-schistosomal target discovery: large-scale screening in S. mediterranea and in vivo candidate validation using vivo-morpholinos. Collectively, our studies will attempt to fill important gaps in functional genomic studies of parasitic flatworms and have the potential to identify new therapeutic targets in Schistosoma. PUBLIC HEALTH RELEVANCE: These studies are pertinent to public health since they aim to develop novel ways to understand the function of genes in the human pathogen Schistosoma mansoni. Completion of the proposed research could aid in the discovery of new drugs to combat disease caused by this important human parasite.