Project description:This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ Onchocerca volvulus is a filarial nematode parasite of humans, causing Onchocerciasis, or River Blindness, which affects over 37 million people, mainly in Africa. It is a severely debilitating disease, which is transmitted to humans by black fly. This project aims to undertake high-throughput sequencing of Onchocerca volvulus transcriptome for de novo assembly of transcripts. The main objective of this project is to recognize genes expressed in given life stages.
Project description:Onchocerca volvulus is the agent of onchocerciasis or river blindness and targeted by WHO for elimination. The main strategy is mass drug administration with ivermectin for the duration of the life span of the adult worms. The drug is mainly microfilaricidal, but weak macrofilaricidal effects, after several rounds of treatment, were observed. A small percentage of adult O. volvulus females contain pleomorphic neoplasms and the rate of worms with neoplasms increases with the number of ivermectin treatments. Within a clinical trial of drug combinations that included ivermectin we identified 24 (5.6%) adult females with neoplasms. We assessed the protein inventory of these neoplasms to identify proteins that may be associated with tumor development. We used laser capture microdissection and highly sensitive mass spectrometry analysis to determine the protein inventory of neoplasms. Neoplasm tissue from female worms was analyzed, and compared to normal tissue from the body wall, uterus and intestine from the same worms, and to tissues from females without neoplasms. One female with neoplasms had a few normal stretched microfilariae in the vagina, but no normal embryogenesis was seen in the other two worms. The healthy females showed all intact embryogenesis. In the worms with neoplasms, 151 proteins were detected in the body wall, 215 proteins in the intestine, 47 proteins in the uterus and 1,577 proteins in the neoplasms. Only in the uterus of the female with intrauterine microfilariae an elevated number of proteins (601) was detectable, while in the uteri of the healthy females 1,710 proteins were detected. The majority of the 20 most abundant proteins identified in neoplasms was highly conserved and only two nematode specific proteins were observed. Among the most abundant proteins that were found in neoplasms but not in the other analyzed tissues, were peroxiredoxin, proteases, and proteins related to signal transduction, ribosomal and proteasome activity. Immunolocalization of a calcium binding protein that was detected in neoplasms confirmed the mass spectrometry results. In conclusion, we have developed a system to analyze the proteome of O. volvulus from nodule sections and identified proteins that are potentially linked to the development of neoplasms and may contribute to worm mortality.
Project description:We report the presence of Onchocerca ochengi and O. volvulus derived small RNAs in bovine nodule fluids and human serum and plasma, respectively. Further comparisons with other related filarial nematodes like Litomosoides sigmodontis and Dirofilaria immitis reveal common and distictive signatures associated to the Onchocerca species.
Project description:We report the presence of Onchocerca ochengi and O. volvulus derived small RNAs in bovine nodule fluids and human serum and plasma, respectively. Further comparisons with other related filarial nematodes like Litomosoides sigmodontis and Dirofilaria immitis reveal common and distictive signatures associated to the Onchocerca species. Examination of small RNA content in bovine nodule fluids and human serum/plasma by Next Generation sequencing