Functional genomics of metacyclogenesis in T.cruzi

描述(申请人提供)：由锥虫引起的恰加斯病 克鲁兹病毒在全球排名第三，仅次于疟疾和血吸虫病 严重的寄生虫病，仍然是中国最严重的寄生虫病 拉丁美洲的残疾调整生命年。尽管最近 控制克氏毛滴虫在残留细菌中传播的进展，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 克氏毛滴虫感染的治疗方法。 后生孢子发生是指非传染性的“昆虫形态” 克鲁兹毛滴虫上胚体分化为侵染性超环体 锥虫。这一过程是科学家们非常感兴趣的话题 因为这对恰加斯病的治疗/预防有影响。我们有 使用了无菌模型系统进行后生周期发生，开始了一个非冗余的 序列验证的cdna序列微阵列，并建立了一个外源 调控表达系统可以用来剖析这一重要的 差异化系统。 项目的具体目标是：1)建立和验证 克氏毛滴虫分化过程中表达基因的综合微阵列 表生鞭毛体转化为准环类鞭毛体。2)使用这些DNA微阵列 找出克氏锥虫在不同时期差异表达的基因 后胚层发育。3)使用已知的克氏锥虫变态周期发生增强剂 确定5-10个触发、控制或指导 分化过程。4)记录这些调控基因的功能 通过计算机分析，过表达，敲除内源基因，或 利用我们的调控T。 CRUZI基因表达系统。 我们预计将鉴定出数百个差异调控基因 特定目标#2。在特定目标#3中，我们将筛选出大多数 在分化过程中没有直接的因果关系，并选择5-10 用于进一步分析的调控基因的候选者。在具体目标4中，我们 将使用我们的可调节的克氏锥虫遗传系统和RNAi来建立 差异表达基因的功能和作用

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