Molecular Definition Of Filarial And Related Nonfilarial Genes And Proteins

丝虫及相关非丝虫基因和蛋白质的分子定义

• 批准号: 10272033
• 负责人: Thomas Nutman
• 金额: $ 83.5万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10272033
• 关键词: Age; Albendazole; Angiostrongylus cantonensis; Ascariasis; Ascaris; Ascaris lumbricoides; Ascaris suum; Biological Markers; Biological Process; Caspase; Code; Complex; Critical Pathways; Culicidae; Data; Development; Diagnostic; Family suidae; Feces; Gene Proteins; Genes; Genetic; Genetic Polymorphism; Genome; Genomics; Goals; Health; Helminths; Heterozygote; Human; Hybrids; Immunoassay; In Vitro; Infection; Inheritance Patterns; Intestinal Volvulus; Knowledge; Larva; Life Cycle Stages; Loa loa; Mass Spectrum Analysis; MicroRNAs; Molecular; Molecular Target; Molting; Mosaicism; Nature; Necator americanus; Nematoda; Nuclear; Onchocerciasis; Parasites; Pathogenicity; Pathway interactions; Patients; Pharmacology; Play; Process; Proteins; Proteome; Proteomics; RNA Interference; Role; Rural; Sampling; Shotguns; Soil; Strongyloides stercoralis; System; Therapeutic Intervention; Time; Trichocephalus trichiura; Wolbachia; base; comparative; endosymbiont; genome analysis; genomic data; gut microbiome; gut microbiota; insight; microbiota; mitochondrial genome; parasitism; programs; protein expression; rapid detection; regional difference; sex; structured data; tool; transcriptomics; transmission process

Having completed the genomes of Loa loa, W. bancrofti, and (most recently) O.volvulus, we have utilized the genomic data as the backdrop for performing a large number of proteomic and transcriptomic studies. We have completed a large-scale proteomic and transcriptomic characterization of almost all the major mammalian stages of O. volvulus, resulting in the identification of more than 85-90% of the products predicted from the O. volvulus genome/putative proteome. The analysis also yielded much of the proteome of Wolbachia, the obligate endosymbiont of O. volvulus. Parasite sex- and stage-specific protein expression identified those pathways related to parasite differentiation. To understand better the developmental programs that underscore the transition between the mosquito-derived infective stage larvae (L3) to mammalian adapted L3s and to L4s following a molt, and the initial week of adaptation to the human host, we adapted an in vitro system that allowed for L3 development and subsequent molting to the L4. Using microarray and proteomic assessments at multiple times through this 9 day process we have not only identified those genes/pathways that are critical for the L3/L4 transition but we have also demonstrated by both pharmacologic inhibition (cysteine protease inhibition) and RNAi (of the critical CPLs) the critical role played by cysteine proteases in the early development of mammalian adapted L3s to L4s. We have recently performed shotgun mass spectroscopy on both human sera of patients with defined filarial infections, excretory/secretory (E/S) products of Loa loa microfilariae, all stages of the O. vovlulus worm, and appropriate controls to identify parasite derived biomarkers of active infection. This has led to identification of molecular targets that have been used d to configure quantitative immunoassays for the rapid detection of active infection for O. volvulus and Loa loa. We examined the role played by soil transmitted helminths in altering the intestinal microbiome by assessing stools collected from residents of 5 rural Kenyan villages prior to and 3 weeks and 3 months following albendazole (ALB) therapy. Interestingly,, the presence of neither Ascaris lumbricoides nor Necator americanus infection significantly altered the overall diversity of the microbiota in comparison with age-matched controls, although Following ALB therapy and clearance of soil-transmitted helminths (STH), there were significant increases in the proportion of the microbiota made up by Clostridiales ( and reductions in the proportion made up by Enterobacteriales. There was also a significant posttreatment decrease in richness, Our data suggest that clearance of STH through deworming alters the gut microbiota. We have generated a 296 megabase (Mb) reference quality genome comprised of 17902 protein-coding genes derived from a single, representative Ascaris worm collected from 60 human hosts in Kenyan villages where pig husbandry is rare. Notably, the majority of human isolates (63/68) possessed mitochondrial genomes that clustered closer to the pig parasite Ascaris suum than to A. lumbricoides. Comparative phylogenomic analyses identified over 11 million nuclear-encoded SNPs but just two distinct genetic types that had recombined across the genomes analysed. The nuclear genomes had extensive heterozygosity and all samples existed as genetic mosaics with either A. suum-like or A. lumbricoides-like inheritance patterns supporting a highly interbred Ascaris species genetic complex. As no barriers appear to exist for anthroponotic transmission of these hybrid worms, a one-health approach to control the spread of human ascariasis will be necessary. MicroRNA for O. volvulus have been identified in the sera of patients with onchocerciasis and regional differences based on miRNA polymorphisms have been identified.

在完成了Loa Loa， W. bancrofti和（最近的）O.volvulus的基因组后，我们利用基因组数据作为进行大量蛋白质组学和转录组学研究的背景。我们已经完成了对O. volvulus几乎所有主要哺乳动物阶段的大规模蛋白质组学和转录组学表征，从而从O. volvulus基因组/推定蛋白质组中鉴定出超过85-90%的预测产物。该分析还获得了沃巴克氏体的大部分蛋白质组，沃巴克氏体是O. volvulus的专性内共生体。寄生虫的性别和阶段特异性蛋白表达确定了这些与寄生虫分化相关的途径。