Novel compounds from predatory assassin bugs as insecticidal and medical purposes [email protected] M. Tonk1*, N. Rügen2*, T. Jenkins3, S. Ainsworth4, H. Vogel5, N. Welsch5, A. Vilcinskas1,2 Deadly straw Assassin bugs are arthropod-feeding predators capable of microinjecting venom and killing their prey within seconds 1 To kill, these bugs: 2 ▪ (1) grab their prey, stab it with a proboscis to inject micro amounts of paralysing and digesting venom ▪ (2) then ingurgitate their pre-digested meal Venomics Assassin bugs (Insecta: Hemiptera: Heteroptera: Reduviidae) possess potent venom used to kill and pre-digest various arthropods as preys. The present study evaluates the insecticidal, cytotoxic and antibacterial effect of crude venom of Rhynocoris iracundus. Strong cytotoxic activity on all tested cell lines and potent antibacterial effect on Escherichia coli was found. To characterise R. iracundus venom in detail and identify the major compounds responsible for its activity, a modern omics-based approach is performed. Identified candidates in the venom will be selected based on their activity and tested for potential agricultural and therapeutic applications. Drugs from bugs – translation of insect toxins into applications The approach includes in vitro assessment of toxicity in different cell types as well as in vivo in insect models Animal Animal Insecticidal activity In-vivo Tenebrio mollitor injection Strong melanisation Organs are digested 99% cytotoxicity Drosophila melanogaster S2 cells Anticancer & Evolutionary antibacterial activities studies 91% cytotoxicity Grant proposals Agriculture mouse neuroblastoma cells Predatory, herbivores, hematophagous 98% cytotoxicity Biomedicine assassin bugs mouse hepatoma cells ► DFG ► TBG-Sequencing Growth curve in presence of venom Dr. Björn M von Reumont E. coli D31 Dr. Heiko Vogel Effect against 1.2 1.0 Max Planck Justus-Liebig Gram (-) E. coli 0.8 0.6 Institute, Jena (600 nm) (600 University, 0.4 Crude venom research Basic Optical density Optical 0.2 Germany Giessen Germany 0 2 4 6 8 10 12 14 16 European assassin bug Time (h) Rhynocoris iracundus Identification of venom compounds Transcriptome analysis An omics-based strategy is used to identify and Proteome analysis pre-characterise new compounds that could be used for medical purposes and crop protection Venom glands Gut Body residues Eggs neurotoxins pore-forming proteins proteases cytolytic proteins disulphide-rich catabolic Venom peptides enzymes Venom glands & gut dissection 9% 8% collection 24% 59% Transcriptome Proteome SDS-PAGE RNA isolation analysis analysis Protein separation Walker et al., 2018; Nat Commun. The assassin bug Pristhesancus plagipennis produces two distinct venoms in separate gland lumens Walker et al., 2017; Mol Cell Proteomics. Melt with this kiss: Paralyzing and liquefying venom of the assassin bug Pristhesancus plagipennis (Hemiptera: Reduviidae) Affiliation 1 Institute for Insect Biotechnology, Justus-Liebig University, Giessen, Germany 2 Fraunhofer IME- BR, Giessen, Germany 3 Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark 4 Alistair Reid Venom Research Unit, Parasitology Department, Liverpool School of Tropical Medicine, Liverpool, United Kingdom 5 Max Planck Institute for Chemical Ecology, Department of Entomology, Jena, Germany.