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.

完成LOA LOA，W。Bancrofti和（最近）O.volvulus的基因组后，我们利用了基因组数据作为进行大量蛋白质组学和转录组研究的背景。 我们已经完成了几乎所有主要的O. volvulus哺乳动物阶段的大规模蛋白质组学和转录表征，从而鉴定了超过85-90％的O. volvulus基因组/推定蛋白质组预测的产物。该分析还产生了沃尔巴奇的大部分蛋白质组，沃尔巴氏菌是O. volvulus的强制性内共生体。 寄生虫性别和特异性蛋白表达鉴定了与寄生虫分化有关的途径。 为了更好地理解强调蚊子衍生的感染性阶段幼虫（L3）对哺乳动物改编的L3和L4之间的过渡，以及对人类宿主的适应的最初一周，我们适应了一个体外系统，可以进行L3的开发，后来摩尔到L4。 在这9天过程中，我们多次使用微阵列和蛋白质组学评估，我们不仅确定了那些对L3/L4过渡至关重要的基因/途径，而且我们还通过药理学抑制（半胱氨酸蛋白酶抑制）和RNAI（CPLS）（CPLS）（CPLS的关键作用）表现出了cysteine Protect in ll3 prodep n ll3 symamm insmamm sommamm sommamm sommamal n amamal nmamm s of n s ramamm insmamm insmamm insmamm insmamm insmamm insmamm insmamal s rabs insmall s rabs insmall s ryamal s ryabs。 最近，我们对具有定义的丝状感染患者的两种人血清进行了shot弹枪质谱法，LOA LOA Microfilariae的排泄/分泌（E/S）产物，O. vovlulus蠕虫的所有阶段以及适当的控制措施，以识别活跃感染的寄生虫衍生的生物标志物。 这导致鉴定了已用于配置定量免疫测定的分子靶标，以快速检测O. volvulus和Loa Loa的主动感染。 我们检查了土壤传播的蠕虫在改变肠道微生物组中发挥的作用，通过评估阿苯达唑（ALB）治疗后的3周和3个月和3个月前的5个肯尼亚村庄的居民收集的粪便。有趣的是，与年龄匹配的对照相比，ass骨lumbricoides和necator Americanus感染都没有显着改变微生物群的总体多样性，尽管在Alb疗法和清除土壤传播的helminths（STH）（STH）的清除之后（STH），通过封闭式质量的比例增加了量的损失（并重新构成）（并重新构成）。我们的数据也大大减少了，我们的数据表明，通过驱虫来清除STH会改变肠道微生物群。 我们已经产生了一个296兆巴（MB）的参考质量基因组，该基因组由17902蛋白质编码基因组成，该基因从单个代表性的asscaris蠕虫中得出，该基因是从肯尼亚村庄的60位人类宿主中收集的，养猪繁殖很少见。值得注意的是，大多数人分离株（63/68）具有与猪寄生虫suum更接近的线粒体基因组，而不是lumbricoides。比较系统生物学分析鉴定出超过1100万个核编码的SNP，但只有两种不同的遗传类型，这些遗传类型已经在分析的基因组中重新组合。核基因组具有广泛的杂合性，并且所有样品都以遗传镶嵌为具有曲霉的植物镶嵌性，均具有曲霉状或曲霉的a。lumbricoides样遗传模式，这些遗传模式支持高度跨性别的阿斯卡里斯物种遗传复合物。由于这些混合蠕虫的拟人化传播似乎不存在障碍，因此需要一种控制人类心脏病传播的一种健康方法。 已经在脑尾尾病患者的血清中鉴定出了O. volvulus的microRNA，并且已经确定了基于miRNA多态性的区域差异。