Function and pharmacology of schistosome multidrug resistance proteins

血吸虫多重耐药蛋白的功能和药理学

• 批准号: 7245276
• 负责人: ROBERT M GREENBERG
• 金额: $ 16.77万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7245276
• 关键词: ABCB1 gene; ATP-Binding Cassette Transporters; Adult; Anthelmintics; Antibodies; Antischistosomal Agents; Binding; Biochemical; Biological Assay; Blood Vessels; Cells; Class; Code; Complementary DNA; Data; Databases; Defect; Development; Disruption; Drug or chemical Tissue Distribution; Drug resistance; Egypt; Excretory function; Exposure to; Fasciola hepatica; Fluorescence; Gene Expression Profile; Gene Frequency; Genes; Genetic; Genome; Goals; Helminths; In Situ Hybridization; Length; Mammalian Cell; Measures; Mediating; Membrane Transport Proteins; Multi-Drug Resistance; Multidrug Resistance Gene; Nematoda; Northern Blotting; Nutrient; P-Glycoprotein; P-Glycoproteins; Parasites; Parasitic Diseases; Pattern; Pharmaceutical Preparations; Pharmacology; Phenotype; Physiological; Physiology; Platyhelminths; Play; Polymerase Chain Reaction; Population; Praziquantel; Praziquantel resistance; Predisposition; Property; Protein Overexpression; Proteins; Publishing; RNA; RNA Interference; Reporting; Research Personnel; Resistance; Role; Schistosoma; Schistosoma mansoni; Schistosomiasis; Schistosomicides; Specificity; Staging; Substrate Specificity; System; Time; Tissues; Toxin; Transmembrane Domain; Tropical Disease; Up-Regulation; Vertebrates; Xenobiotics; cell motility; efflux pump; fluorexon; genetic selection; inhibitor/antagonist; interest; member; multi drug transporter; neoplastic cell; programs; response; vector; wasting

DESCRIPTION (provided by applicant): Project Summary: Trematode flatworms of the genus Schistosoma are the causative agents of schistosomiasis. One potential physiological target for new anti-schistosoma drugs is the worm's system for excretion of wastes and xenobiotics. P-glycoprotein (Pgp), a member of the ATP-binding cassette superfamily of proteins, is an ATP-dependent efflux pump involved in transport of toxins and xenobiotics from cells. In vertebrates, Pgp is the product of the multi-drug-resistance 1 gene, which is amplified and over-expressed in tumor cells that show broad drug resistance. Pgp may also play a role in drug resistance in helminths. A schistosome Pgp cDNA (SMDR2) was sequenced several years ago, but has not been functionally characterized. The long-term goal of this proposal is to dissect the functional role and pharmacological sensitivities of SMDR2 and other schistosome drug transporters. We will adapt a calcein fluorescence assay to examine the substrate and inhibitor specificities of expressed SMDR2 and other transporters. We will also examine the effects of praziquantel, the current antischistosomal of choice, on expression of SMDR2, and whether praziquantel-resistant isolates have altered expression of this or other transporters. Finally, we will examine the effect on the parasite of genetic or pharmacological disruption of SMDR2. The specific aims of the project are to answer the following questions: 1. What are the biochemical properties and substrate specificities of SMDR2 and other schistosome drug transporters expressed in mammalian cells? 2. What is the tissue distribution and developmental profile of SMDR2 and other schistosome multi-drug transporters? 3. Does exposure to agents such as PZQ result in changes in SMDR2 expression or distribution? 4. Do isolates of worms with reduced susceptibility to PZQ show differences in expression levels of SMDR2 or other multidrug transporters? 5. What effects do genetic and pharmacological disruption of SMDR2 have on parasite survival, physiology and pharmacological sensitivities/relevance: Schistosomiasis is a major tropical disease caused by parasitic flatworms called schistosomes. Potential physiological targets for new drugs against schistosomiasis might be the transporters that remove wastes and toxins from schistosome cells. We propose to use several approaches to determine the properties of one such schistosome molecule, P-glycoprotein, which, in vertebrates, is also involved in resistance to a broad array of drugs.

项目简介（由申请人提供）：项目简介：血吸虫属扁虫是血吸虫病的病原体。新的抗血吸虫药物的一个潜在的生理靶点是血吸虫排泄废物和外源性药物的系统。p -糖蛋白（Pgp）是atp结合盒超家族蛋白的一员，是atp依赖的外排泵，参与细胞内毒素和外源物的运输。在脊椎动物中，Pgp是多重耐药1基因的产物，该基因在具有广泛耐药的肿瘤细胞中被扩增和过表达。Pgp也可能在蠕虫的耐药性中起作用。几年前对血吸虫Pgp cDNA （SMDR2）进行了测序，但尚未进行功能表征。本提案的长期目标是剖析SMDR2和其他血吸虫药物转运体的功能作用和药理学敏感性。我们将采用钙黄蛋白荧光法来检测表达的SMDR2和其他转运蛋白的底物和抑制剂的特异性。我们还将研究吡喹酮（目前首选的抗血吸虫药）对SMDR2表达的影响，以及吡喹酮耐药分离株是否改变了该转运体或其他转运体的表达。最后，我们将研究SMDR2基因或药理学破坏对寄生虫的影响。该项目的具体目的是回答以下问题：1。SMDR2和其他血吸虫药物转运体在哺乳动物细胞中表达的生化特性和底物特异性是什么？2. SMDR2和其他血吸虫多药转运体的组织分布和发育概况是什么？3. 暴露于诸如PZQ之类的药物会导致SMDR2表达或分布的改变吗？4. 对PZQ敏感性降低的蠕虫分离株在SMDR2或其他多药转运蛋白的表达水平上是否存在差异？5. SMDR2基因和药理学破坏对寄生虫存活、生理和药理学敏感性/相关性的影响：血吸虫病是由称为血吸虫的寄生扁形虫引起的一种主要热带疾病。从血吸虫细胞中清除废物和毒素的转运体可能是抗血吸虫病新药的潜在生理靶点。我们建议使用几种方法来确定一种这样的血吸虫分子p -糖蛋白的特性，在脊椎动物中，p -糖蛋白也参与对多种药物的抗性。