In vitro silencing of Brugia malayi trehalose-6-phosphate phosphatase impairs embryogenesis and in vivo development of infective larvae in jirds.
In vitro silencing of Brugia malayi trehalose-6-phosphate phosphatase impairs embryogenesis and in vivo development of infective larvae in jirds.
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DOI：
10.1371/journal.pntd.0001770
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发表时间：
2012
影响因子：
3.8
通讯作者：
Bhattacharya SM
中科院分区：
文献类型：
--
作者：
Kushwaha S;Singh PK;Shahab M;Pathak M;Bhattacharya SM
由于海藻糖在胚胎发育、葡萄糖吸收和抗应激中的重要作用，海藻糖代谢酶在分枝杆菌、植物线虫、昆虫和真菌等生物中被认为是潜在的药物或疫苗靶标。海藻糖-6-磷酸磷酸酶(TPP)是哺乳动物体内尚未见报道的海藻糖生物合成酶之一。秀丽线虫中TPP基因的沉默揭示了TPP在线虫中不可或缺的功能。在本研究中，通过siRNA介导的沉默研究了马来丝虫TPP基因的功能，进一步证实了该酶是一个潜在的抗丝虫病药物靶点。在成年雌性马来丝虫体内，TPP基因的沉默导致了严重的表型畸形，如畸形和胚胎发育停滞。寄生虫的活动能力显著降低，雌虫体内微丝虫的产生及其体外释放也大大减少。大多数释放到培养基中的微丝虫被发现死亡。感染马来丝虫的幼虫经TPP基因沉默后，幼稚沙鼠腹膜腔内接种成虫后，84.9%的成虫建立减少。本研究结果提示，马来丝虫TPP在雌虫胚胎发生、幼虫感染性和寄生虫生存中起重要作用。因此，马来丝虫的TPP酶具有作为抗丝虫病药物靶点的潜力。淋巴丝虫病是一种被忽视的热带疾病，是导致永久性和长期残疾的第二大原因。这种疾病的控制依赖于大量给药，这些药物主要作用于微丝虫，而对成虫没有实质性影响。需要持续几年的药物来阻止感染的传播，这可能会导致抗药性寄生虫的发展。海藻糖在线虫体内发挥着重要的作用，海藻糖生物合成酶被认为是药物或疫苗候选的潜在靶点。在本研究中，我们沉默了海藻糖-6-磷酸磷酸酶，并研究了TPP酶在丝虫马来丝虫存活、雌虫胚胎发生和宿主感染建立中的生物学功能。用5 mM浓度的siRNA对成虫进行基因沉默，用2 mM浓度的siRNA处理L3，再接种沙鼠腹膜腔，研究siRNA处理对体内幼虫发育的影响。目前的发现证实了海藻糖-6-磷酸磷酸酶是一个重要的抗丝虫病药物靶点。
The trehalose metabolic enzymes have been considered as potential targets for drug or vaccine in several organisms such as Mycobacterium, plant nematodes, insects and fungi due to crucial role of sugar trehalose in embryogenesis, glucose uptake and protection from stress. Trehalose-6-phosphate phosphatase (TPP) is one of the enzymes of trehalose biosynthesis that has not been reported in mammals. Silencing of tpp gene in Caenorhabditis elegans revealed an indispensable functional role of TPP in nematodes. In the present study, functional role of B. malayi tpp gene was investigated by siRNA mediated silencing which further validated this enzyme to be a putative antifilarial drug target. The silencing of tpp gene in adult female B. malayi brought about severe phenotypic deformities in the intrauterine stages such as distortion and embryonic development arrest. The motility of the parasites was significantly reduced and the microfilarial production as well as their in vitro release from the female worms was also drastically abridged. A majority of the microfilariae released in to the culture medium were found dead. B. malayi infective larvae which underwent tpp gene silencing showed 84.9% reduced adult worm establishment after inoculation into the peritoneal cavity of naïve jirds. The present findings suggest that B. malayi TPP plays an important role in the female worm embryogenesis, infectivity of the larvae and parasite viability. TPP enzyme of B. malayi therefore has the potential to be exploited as an antifilarial drug target. Lymphatic filariasis, one of the neglected tropical diseases, is the second leading cause of permanent and long term disability. Control of the disease relies on the mass administration of drugs which mainly act on the microfilariae without substantial effect on adult worms. Drugs need to be continued for several years to block the transmission of infection which may result in to development of resistant parasites. The sugar trehalose has been shown to play several important functions in the nematodes, and trehalose biosynthetic enzymes have been considered as potential targets for drug or vaccine candidate. In the present study we silenced trehalose-6-phosphate phosphatase and studied the biological function of TPP enzyme in the filarial nematode B. malayi viability, female worm embryogenesis and establishment of infection in the host. In vitro gene silencing was done in adult parasites using 5 mM concentration of siRNA while 2 mM of siRNA was used to treat L3 which were further inoculated into the peritoneal cavity of jirds to study the effect of siRNA treatment on in vivo larval development. The present findings validate trehalose-6-phosphate phosphatase as a vital antifilarial drug target.
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影响因子：
1.3
作者：
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通讯作者：
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影响因子：
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Talbot, NJ
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Ghedin, Elodie;Wang, Shiliang;Scott, Alan L.
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Scott, Alan L.