Schistosomiasis is a tropical parasitic disease infecting over 200 million people. Treatment relies on a single drug, praziquantel (PZQ). In the absence of back-up drugs with PZQ's therapeutic spectrum, the risk of resistance to PZQ and eventual drug failure is a major concern. Traditional phenotypic screens, using adult- stage S. mansoni, are low-throughput and incompatible with modern high-throughput screen (HTS) systems. In keeping with the NIAID's mission, the present proposal aims to turn a newly developed, moderate- throughput phenotypic screen (MTS), into a fully automated, quantitative HTS to accelerate drug discovery for this infectious disease. The proposal involves three PIs with ongoing collaborations and respective biological, screening-technology and bio-computational skills who are focused on just this goal. As a first research track for this proposal, we will utilize in-house automation and a high-content screening (HCS) system to significantly increase throughput over our published MTS approach. The proposal will involve;expanding robotic plating of the parasite, developing protocols for bright-field and fluorescence-based microscopy and adapting commercial image-analysis software to identify (segment), quantitatively describe and track the motion of parasites with a view to prioritizing compounds for further pre-clinical development. Because commercial HCS analysis tools are not likely optimized for recording the complex and dynamic phenotypes displayed by this multicellular parasite, we will also pursue a second and parallel track of research. Specifically, we will develop de novo an automated image-analysis screening technology to define, identify, and quantify the range of phenotypic responses (morphological and behavioral) possible in this parasite. Ultimately, both the experimental and computation tracks will together produce a standardized HTS protocol and a comprehensive, quantitative suite of image-analysis programs to categorize parasite phenotypes. Such rigor will facilitate the screening of large numbers of potential compounds and their prioritization into the secondary and tertiary screening assays available in-house. We also intend to make the algorithmic framework including its methods and implementations, publicly available.
The major goal of the project is to turn a moderate-throughput phenotypic screen (MTS) system for schistosomiasis, into a fully automated, quantitative high-throughput screen (HTS). By so doing, the rate of discovery of drugs to treat this global tropical disease will be increased.
|Long, Thavy; Neitz, R Jeffrey; Beasley, Rachel et al. (2016) Structure-Bioactivity Relationship for Benzimidazole Thiophene Inhibitors of Polo-Like Kinase 1 (PLK1), a Potential Drug Target in Schistosoma mansoni. PLoS Negl Trop Dis 10:e0004356|
|DvoÅ™Ã¡k, Jan; FajtovÃ¡, Pavla; UlrychovÃ¡, Lenka et al. (2016) Excretion/secretion products from Schistosoma mansoni adults, eggs and schistosomula have unique peptidase specificity profiles. Biochimie 122:99-109|
|Glaser, Jan; Schurigt, Uta; Suzuki, Brian M et al. (2015) Anti-Schistosomal Activity of Cinnamic Acid Esters: Eugenyl and Thymyl Cinnamate Induce Cytoplasmic Vacuoles and Death in Schistosomula of Schistosoma mansoni. Molecules 20:10873-83|
|Fonseca, Nayara Cristina; da Cruz, Luana Faria; da Silva Villela, Filipe et al. (2015) Synthesis of a sugar-based thiosemicarbazone series and structure-activity relationship versus the parasite cysteine proteases rhodesain, cruzain, and Schistosoma mansoni cathepsin B1. Antimicrob Agents Chemother 59:2666-77|
|Asarnow, Daniel; Rojo-Arreola, Liliana; Suzuki, Brian M et al. (2015) The QDREC web server: determining dose-response characteristics of complex macroparasites in phenotypic drug screens. Bioinformatics 31:1515-8|
|Rojo-Arreola, Liliana; Long, Thavy; Asarnow, Dan et al. (2014) Chemical and genetic validation of the statin drug target to treat the helminth disease, schistosomiasis. PLoS One 9:e87594|
|Shimoide, Alan; Kimball, Ian; Gutierrez, Alba A et al. (2013) Quantification and analysis of ecdysis in the hornworm, Manduca sexta, using machine vision-based tracking. Invert Neurosci 13:45-55|
|Asarnow, Daniel E; Singh, Rahul (2013) Segmenting the etiological agent of schistosomiasis for high-content screening. IEEE Trans Med Imaging 32:1007-18|