HIV-1 Integrates Widely throughout the Genome of the Human Blood Fluke Schistosoma mansoni.
HIV-1 Integrates Widely throughout the Genome of the Human Blood Fluke Schistosoma mansoni.
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DOI：
10.1371/journal.ppat.1005931
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发表时间：
2016-10
期刊：
影响因子：
6.7
通讯作者：
Brindley PJ
中科院分区：
文献类型：
--
作者：
Suttiprapa S;Rinaldi G;Tsai IJ;Mann VH;Dubrovsky L;Yan HB;Holroyd N;Huckvale T;Durrant C;Protasio AV;Pushkarsky T;Iordanskiy S;Berriman M;Bukrinsky MI;Brindley PJ
就发病率和死亡率而言，血吸虫病是人类最重要的蠕虫病。使用慢病毒对核糖体的简易操作将使这些和相关被忽视的热带疾病病原体（包括绦虫）及其非分裂细胞的功能基因组学取得进展。迄今为止，还没有这种办法。用水泡性口炎病毒糖蛋白假型化的人HIV-1分离株NL 4 -3感染人血吸虫（Schistosomamansoni）的血流型。强终止子和正链cDNA的出现表明病毒粒子融合到溶酶体细胞，核衣壳内化，RNA基因组逆转录。锚定PCR分析、测序HIV-1特异性锚定Illumina文库和染色体的全基因组测序（WGS）证实了染色体整合; > 8，000个整合被定位，分布在包括性染色体在内的8对染色体中。基因组中的整合率超过每1，000 kb 5个，HIV-1整合到蛋白质编码基因座和其他地方，整合偏好与人类T细胞不同。基于WGS，我们估计每个染色体胚泡约有2，100个整合，即每个细胞约有两个或三个事件，与人类细胞中的整合率相当。考虑到慢病毒属的天然宿主--灵长类动物和慢病毒之间的系统发育距离，在慢病毒中完成了HIV-1复制所必需的进入后过程，包括整合酶催化的整合，这是非常显著的。这些神秘的发现揭示了HIV-1在S. mansoni，并首次证明HIV-1可以整合到无脊椎动物的基因组中。血吸虫病是一种主要的被忽视的热带疾病，影响着发展中国家的2亿多人。寄生虫的基因组序列已经被解码;它包括> 10，000个基因。寻找控制这种NTD的新方法，基因组信息可能为新的治疗提供靶点。确定特定基因的作用和重要性的方法将有助于这些任务。逆转录病毒HIV-1是艾滋病毒/艾滋病的病原体，已被广泛研究和改造，用于生物医学研究。使用实验室修饰形式的HIV-1，我们操纵了曼氏血吸虫（Schistosoma mansoni）的基因组，曼氏血吸虫是血吸虫的主要物种之一。实验室修饰的HIV-1感染了染色体，并插入了寄生虫的染色体。使用下一代测序对这些染色体插入进行定位，并分布在包括性染色体在内的整个染色体上。这些发现是值得注意的，因为他们揭示了HIV-1在S. mansoni，他们提供了第一个证明，HIV-1可以整合到无脊椎动物的基因组。他们为研究血吸虫病的新疗法铺平了道路。
Schistosomiasis is the most important helminthic disease of humanity in terms of morbidity and mortality. Facile manipulation of schistosomes using lentiviruses would enable advances in functional genomics in these and related neglected tropical diseases pathogens including tapeworms, and including their non-dividing cells. Such approaches have hitherto been unavailable. Blood stream forms of the human blood fluke, Schistosoma mansoni, the causative agent of the hepatointestinal schistosomiasis, were infected with the human HIV-1 isolate NL4-3 pseudotyped with vesicular stomatitis virus glycoprotein. The appearance of strong stop and positive strand cDNAs indicated that virions fused to schistosome cells, the nucleocapsid internalized and the RNA genome reverse transcribed. Anchored PCR analysis, sequencing HIV-1-specific anchored Illumina libraries and Whole Genome Sequencing (WGS) of schistosomes confirmed chromosomal integration; >8,000 integrations were mapped, distributed throughout the eight pairs of chromosomes including the sex chromosomes. The rate of integrations in the genome exceeded five per 1,000 kb and HIV-1 integrated into protein-encoding loci and elsewhere with integration bias dissimilar to that of human T cells. We estimated ~ 2,100 integrations per schistosomulum based on WGS, i.e. about two or three events per cell, comparable to integration rates in human cells. Accomplishment in schistosomes of post-entry processes essential for HIV-1replication, including integrase-catalyzed integration, was remarkable given the phylogenetic distance between schistosomes and primates, the natural hosts of the genus Lentivirus. These enigmatic findings revealed that HIV-1 was active within cells of S. mansoni, and provided the first demonstration that HIV-1 can integrate into the genome of an invertebrate. Schistosomiasis is a major neglected tropical disease (NTD), which afflicts > 200 million people in developing countries. The genome sequence of the schistosome parasite has been decoded; it includes > 10,000 genes. New approaches to control this NTD are sought and genomic information may provide targets for new treatments. Methods to determine the role and importance of specific genes would facilitate these tasks. The retrovirus HIV-1, the causative agent of HIV/AIDS, has been extensively studied and modified for use in biomedical research. Using a lab-modified form of HIV-1, we manipulated the genome of Schistosoma mansoni, one of the major species of schistosomes. Lab-modified HIV-1 infected schistosomes and inserted in the chromosomes of the parasite. These chromosomal insertions were mapped using next generation sequencing and were distributed throughout the chromosomes including the sex chromosomes. The findings were notable since they revealed that HIV-1 was active within cells of S. mansoni, and they provide the first demonstration that HIV-1 can integrate into the genome of an invertebrate. They pave a route forward for investigating new therapies for schistosomiasis.
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影响因子：
3.8
作者：
Bagasra, O;Lavi, E;Pomerantz, RJ
通讯作者：
Pomerantz, RJ
影响因子：
4.8
作者：
Faghiri, Zahra;Skelly, Patrick J.
通讯作者：
Skelly, Patrick J.
影响因子：
6.7
作者：
Engelman A;Cherepanov P
通讯作者：
Cherepanov P
影响因子：
3.7
作者：
Engelman, Alan;Cherepanov, Peter
通讯作者：
Cherepanov, Peter
DOI：
10.1016/j.ijpddr.2014.09.007
发表时间：
2014-12
影响因子：
4
作者：
Greenberg, Robert M.
通讯作者：
Greenberg, Robert M.