In vitro silencing of Brugia malayi trehalose-6-phosphate phosphatase impairs embryogenesis and in vivo development of infective larvae in jirds.
BACKGROUND
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.


RESULTS
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.


CONCLUSIONS
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.
Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
Susheela
S
Kushwaha
Prashant Kumar
PK
Singh
Mohd
M
Shahab
Manisha
M
Pathak
Shailja Misra
SM
Bhattacharya
D000818:Animals; D017178:Brugia malayi / Q000201:enzymology* / Q000254:growth & development* / Q000472:pathogenicity; D004195:Disease Models, Animal; D005260:Female; D005368:Filariasis / Q000469:parasitology* / Q000473:pathology; D020868:Gene Silencing; D005849:Gerbillinae / Q000469:parasitology; D007814:Larva / Q000201:enzymology / Q000254:growth & development / Q000472:pathogenicity; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism
D034741:RNA, Small Interfering; C023178:trehalose-phosphatase; D010744:Phosphoric Monoester Hydrolases