Functional genomics of metacyclogenesis in T.cruzi

描述（由申请人提供）：锥虫病，由锥虫属引起 克鲁兹是全球仅次于疟疾和血吸虫病的第三大流行病， 严重的寄生虫病，仍然是最严重的寄生虫病， 拉丁美洲按残疾调整生命年计算的情况。尽管最近 控制T. cruzi在reduviid错误，没有治疗 可用于慢性恰加斯病，而急性感染只能 用剧毒药物治疗因此，迫切需要新的 治疗T.克鲁兹感染。 后气旋发生是指非传染性的“昆虫形态” epimastigotes of T.克鲁兹分化成传染性亚循环 锥鞭毛体这个过程是科学家们非常感兴趣的课题 因为这对治疗/预防南美锥虫病有重要意义。我们有 使用了一个纯生的模式系统的元气旋，开始了一个非冗余的 微阵列的序列验证的cDNA序列，并建立了一个外源 调控表达系统，可用于剖析这一重要的 分化系统 本项目的具体目标是：1）建立和验证 T. cruzi基因在分化过程中表达 上鞭毛体转化为后环锥鞭毛体。2)使用这些DNA微阵列 鉴定T. cruzi基因的差异表达， 后气旋生成3)利用T.克鲁济变质旋回作用 确定5-10个候选基因，触发，控制或指导 分化过程。4)为了记录这些调控基因的功能 通过计算机模拟分析、过表达、敲除内源基因，或 表达的调节抑制（例如，使用RNAi），使用我们调节的T. cruzi基因表达系统。 我们预计在这些基因中发现几百个差异调节基因。 具体目标#2在具体目标#3中，我们将筛选出大多数基因， 在分化过程中不是直接因果关系，选择5-10 用于进一步分析的调节基因的候选物。在具体目标#4中， 将使用我们的可调节T。cruzi基因系统和RNAi技术， 差异表达基因的功能和作用，发挥可能的

DESCRIPTION (provided by applicant): Chagas disease, caused by Trypanosoma cruzi, is globally ranked behind malaria and schistosomiasis as the third most serious parasitic disease, and remains the most serious parasitic disease in Latin America in terms of disability adjusted life years. Despite recent progress controlling dissemination of T. cruzi in reduviid bugs, no treatment is available for chronic Chagas disease, and acute infections can only be treated with highly toxic drugs. Thus, there is a critical need for new approaches to treatment of T. cruzi infections. Metacyclogenesis is the process by which non-infectious "insect form" epimastigotes of T. cruzi differentiate into infectious metacyclic trypomastigotes. This process is the subject of much interest among scientists because of the implications for treatment/prevention of Chagas disease. We have used an axenic model system for metacyclogenesis, begun a nonredundant microarray of sequence-verified cDNA sequences, and established an exogenously regulated expression system that can be used to dissect this important differentiation system. The specific aims of the project are: 1) To establish and validate comprehensive microarrays of T. cruzi genes expressed during differentiation of epimastigotes into metacyclic trypomastigotes. 2) To use these DNA microarrays to identify T. cruzi genes that are differentially expressed during metacyclogenesis. 3) To use known potentiators of T. cruzi metacyclogenesis to identify 5-10 candidate genes that trigger, control or direct the differentiation process. 4) To document the function of these regulatory genes by in silico analysis, overexpression, knockout of the endogenous genes, or regulated inhibition of expression (e.g., using RNAi), using our regulated T. cruzi gene expression system. We anticipate identifying several hundred differentially regulated genes in Specific Aim #2. In Specific Aim #3, we will screen out most of the genes that are not directly causal in the differentiation process and select 5-10 candidates for regulatory genes for further analysis. In Specific Aim #4, we will use our regulatable T. cruzi genetic system and RNAi to establish the functions and roles of the differentially expressed genes that play a probable