Global gene expression analysis of Trypanosoma cruzi under hyperosmotic stress

描述(由申请人提供)：这项研究将主要在阿根廷圣马丁，圣马丁国立大学生物技术研究所，与Juan J.Cazzulo博士合作，作为NIH补助金1R01 AI068647的延伸。恰加斯病的化疗仅限于毒性较高、疗效有限的药物。我们的研究集中于通过研究寄生虫所必需的代谢系统来合理寻找有效的化疗治疗方法，但在人类宿主中没有同等的代谢系统。寄生虫对营养和高渗应激的反应是其在媒介和哺乳动物宿主中生存的基础，它的研究可能导致发现新的靶点。应激条件通常会导致细胞水平上的各种生理反应。应激条件通常会导致细胞水平上的各种生理反应。当克鲁兹毛虫的上胚体受到高渗胁迫时，细胞体积缩小而没有恢复，并上调或下调了一些基因的表达。在几个转录本的3‘-UTR中发现了候选的结构RNA基序，当与完整的转录组进行比较时，这些候选结构RNA基序在实验数据集中得到了丰富。这些顺式元件可能参与对高渗透胁迫做出反应的整体表达模式的转录后机制。具体目标1：研究高渗胁迫下克氏木霉基因表达的变化；具体目标2：研究受高渗胁迫影响的基因3‘-UTR区中的顺式元件在克氏木霉基因表达中的作用。 公共卫生相关性：在美洲，从北部的墨西哥到南部的阿根廷和智利，有1600万至1900万人感染了克氏锥虫，这是恰加斯病的病原体。据估计，每年的发病率为561,000例。恰加斯病是一种媒介传播的疾病，也可以通过输血传播，是拉丁美洲充血性心力衰竭的主要原因。我们的目标是找到干扰克氏锥虫代谢途径的方法，作为控制这种寄生虫和类似寄生虫引起的感染的策略。克氏毛滴虫对营养和高渗胁迫的反应与哺乳动物细胞对类似应激的反应不同，他们的研究可能会导致发现新的杀锥虫剂靶标。这项工作旨在研究这些途径在克氏锥虫中的作用和意义。

DESCRIPTION (provided by applicant): This research will be done primarily in San Martin, Argentina, at Fundacion Instituto de Investigaciones Biotecnologicas, Universidad Nacional de General San Martin, in collaboration with Dr. Juan J. Cazzulo, as an extension of NIH Grant 1R01 AI068647. Chemotherapy of Chagas disease is restricted to drugs with relatively high toxicity and limited efficacy. Our research focuses on the rational search for effective chemotherapeutic treatments for T. cruzi by investigating metabolic systems necessary for parasites but without equivalent counterparts in the human host. The response of the parasite to nutritional and hyperosmotic stresses is fundamental for its survival within the vector and mammalian hosts and its study could lead to the finding of novel targets. Stress conditions usually lead to a variety of physiological responses at the cellular level. Stress conditions usually lead to a variety of physiological responses at the cellular level. When epimastigotes of T. cruzi were submitted to hyperosmotic stress the cells reduced their volume without recovery, and up-regulated or down- regulated the expression of a number of genes. Candidate structural RNA motifs were found in the 3'-UTR of several transcripts and enriched in the experimental dataset when comparing over the complete transcriptome. These cis-elements could be involved in post-transcriptional mechanisms underlying a global expression pattern in response to hyperosmotic stress. This proposal will be focused in the following specific aims: Specific aim 1: to study the changes in gene expression in T. cruzi submitted to hyperosmotic stress; Specific aim 2: to investigate the role of the cis-elements identified in the 3'-UTR of genes affected by hyperosmotic stress in gene expression of T. cruzi. PUBLIC HEALTH RELEVANCE: In the Americas, from Mexico in the North to Argentina and Chile in the South, there are 16 to 19 million people infected with Trypanosoma cruzi, the causative agent of Chagas disease. Estimated yearly incidence amounts to 561,000 cases. Chagas disease is a vector-borne disease that can also be transmitted by blood transfusion and is the leading cause of congestive heart failure in Latin America. Our goal is to find ways of interfering with Trypanosoma cruzi metabolic pathways as a strategy of controlling infections caused by this and similar parasites. The response of T. cruzi to nutritional and hyperosmotic stresses is different from the response of mammalian cells to similar stresses and their study may lead to the discovery of new targets for trypanocidal agents. This work is designed to investigate the roles and significance of these pathways in T. cruzi.