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: 7649449
• 负责人: ROBERT M GREENBERG
• 金额: $ 34.76万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 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 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649449
• 关键词: ABCB1 gene; ATP-Binding Cassette Transporters; Adult; Anthelmintics; Antibodies; Antischistosomal Agents; Binding; Biochemical; Biological Assay; Blood Vessels; Cells; Code; Complementary DNA; Data; Databases; Defect; Development; 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; Population; Praziquantel; Praziquantel resistance; Predisposition; Property; 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; overexpression; 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.

描述（由申请人提供）：项目摘要：血吸虫属的Trematode Flatems是血吸虫病的致病药物。新型抗刺激性药物的潜在生理靶标是蠕虫排泄废物和异种生物的系统。 P-糖蛋白（PGP）是ATP结合盒的蛋白质超家族的成员，是ATP依赖的外排泵，参与了来自细胞的毒素和异种生物的转运。在脊椎动物中，PGP是多药耐药1基因的产物，该基因在表现出广泛耐药性的肿瘤细胞中被扩增和表达。 PGP也可能在舵机中的耐药性中发挥作用。几年前测序了一个血吸虫PGP cDNA（SMDR2），但尚未在功能上表征。该提案的长期目标是剖析SMDR2和其他SCHISTOSOM药物转运蛋白的功能作用和药理敏感性。我们将适应钙调蛋白荧光测定法，以检查表达SMDR2和其他转运蛋白的底物和抑制剂特异性。我们还将研究Praziquantel，当前选择的反静脉体对SMDR2表达的影响，以及抗胰烷耐药的分离株是否改变了该转运蛋白的表达。最后，我们将研究对SMDR2遗传或药理破坏的寄生虫的影响。该项目的具体目的是回答以下问题：1。哺乳动物细胞中表达的SMDR2和其他SMDR2和其他螺旋体药物转运蛋白的生化特性和底物特异性是什么？ 2。SMDR2和其他SCISTOSOM多药物转运蛋白的组织分布和发育特征是什么？ 3。暴露于PZQ等药物会导致SMDR2表达或分布的变化？ 4。对PZQ敏感性降低的蠕虫的分离株在SMDR2或其他多药转运蛋白的表达水平上差异吗？ 5。SMDR2对遗传学和药理破坏对寄生虫生存，生理和药理敏感性/相关性有什么影响：血吸虫病是由称为血吸虫的寄生虫引起的主要热带疾病。针对血吸虫病的新药物的潜在生理靶标可能是从螺构细胞中去除废物和毒素的转运蛋白。我们建议使用几种方法来确定一种这种黑螺链体分子P-糖蛋白的性质，该分子在脊椎动物中也参与了对广泛药物的抗性。