Tracking toxins in venomous Vermes: comparative venomics of polychaete annelids

Lead Research Organisation: The Natural History Museum
Department Name: Life Sciences

Abstract

Although most lay people have little problem in identifying creatures like jellyfish, wasps, spiders, and scorpions as potentially dangerous venomous organisms, far fewer know that iconic animals such as the Komodo dragon, or the inconspicuous centipedes possess potent venoms. Similarly, although digging up bloodworms-a group of marine polychaete annelids-for use as fish bait is a multi-million dollar cottage industry along the coasts of the Northeastern United States and Canada, it is doubtful whether unsuspecting fishermen realize that they are hooking worms that possess potent neurotoxic venom. Bloodworms and several groups of the related scale-worm have two pairs of hardened jaw, associated with venom gland. Although these venoms presumable play a crucial role in the life of the animals in subduing prey, we know almost nothing about the make-up of their venoms. This leaves not only a large hole in our understanding of an important group of organisms-representing possibly as many as 1,000 species of venomous worms-but it also compromises our general understanding of venom biology and evolution in the animal kingdom. This study aims to remedy this ignorance by performing the first extensive and intensive analysis of the composition of polychaete venoms.

This project uses a combination of cutting edge techniques to determine venom toxin composition on both the genetic and protein levels. Eight species of worms have been selected-4 species of bloodworms and 4 species of scale-worms-to represent a cross section of the major groups they represent. For each species, up to half a million mRNA sequences will be characterized. These precursor molecules are the templates for the production of toxin proteins. In addition, the toxin proteins themselves will also be determined directly from extracted venom. Together these data will reveal what toxins are expressed in the venom glands of the worms, providing an important contribution towards elucidating the precise identity of one toxin that is known to exist in bloodworm venom. This is important because this toxin has a very specific mode of action-paralyzing and killing crustacean prey in the wild-that is already of use to researchers who study neuronal processes in the laboratory.

By comparing venom toxins between different groups of worms and other venomous animals we can also start to address important questions relating to the evolution of venoms and venomous organisms. For example, venoms are a prime example of convergent evolution, the repeated independent evolution of similar adaptations in different taxa. Since venoms have evolved many times independently, a comparison of their composition can reveal shed light on the rules under which convergent evolution operates.

Lastly, the outstanding feature of venoms, and the source of our fascination with them, is of course the physiological effects they have. This project will test specific types of pharmacological activities of the worm venoms, such as neurotoxity. This will lead to a better understanding of the biology of the organisms, but perhaps even more importantly, understanding the bioactivities of toxins can have valuable downstream applications. Already seven prescription drugs have been developed from components of animal venoms, from animals as different as cone snails and the Gila monster. Since these are used to treat serious conditions in humans, including diabetes and hypertension, potentially many people could benefit from mining the hidden biodiversity of venoms.

Technical Summary

Identify the toxins expressed in the venom glands of 8 species of polychaete annelids
- Use 454 GS FLX Titanium technology to generate the first comprehensive polychaete venom gland transcriptomes
- Use nanoLC-MS/MS to identify toxin proteins to generate the first comprehensive polychaete proteome data
- Compare the transcriptome and proteome data to shed light on the poorly understood relationship between venom gland transcriptomes and proteomes
- Use the new data as an important step towards elucidation of the complete sequence of alpha-glycerotoxin, an unusual and useful neurotoxin from bloodworms

Understand whether more close related species convergently recruit venom toxins from more similar toxin families than distantly related species
- Use standard phylogenetic methods (principally model-based methods such as Bayesian analysis and ML) to estimate toxin gene trees including the polychaete toxins and those of other studied venomous animals to produce the first explicit assessment of the role of phylogeny in shaping venom composition across phylogenetic levels ranging from homologous venoms in closely related species to convergent venoms in species of more or less closely related families to convergent venoms between phyla and major bilaterians clades
- Use the new data and resulting trees to re-assess current hypotheses about the evolution of toxin gene families across Metazoa, for example the importance the importance of gene duplications in contributing to venom complexity.

Reveal the pharmacological activities of polychaete venoms
- Use enzymatic and non-enzymatic in vitro and in vivo essays to obtain a better understanding of the toxic activities of bloodworm venoms, and generate the first data on venom activity in scale-worms

Planned Impact

Because venomics is richly interdisciplinary in terms of its concepts, methods, data and the nature of the questions it addresses, this project should be of interest to the wider community of biologists. Also, because venoms have proven to be rich natural resources for bioactive compounds that have been used for developing commercially successful drugs, venom research is very interesting to the pharmaceutical industry. Moreover, venomous animals have long played important roles in cultures the world over, and they are widely used in traditional medicine. As reservoirs of a great diversity of useful bioactive compounds venomous animals are therefore medically and economically valuable natural resources. The first step in safeguarding the medical and economic potential of native venomous species is to understand what bioactive compounds are present in their venoms, and what pharmacological effects these may have, which is the project's explicit focus. Therefore, the conservation and protection of native venomous species in the wild deserves special attention. As an illustration of the need for this, the harvesting of the bloodworm Glycera dibranchiata is a multi-million dollar business along the coasts of the United States and Canada. The worms are sold as bait, and are exported also to Europe, often as live individuals. This introduces the risk that such species establish themselves as invasive species, which may have detrimental effects on native populations of close related European species. Finally, venomous animals have a unique 'cool' factor for general audiences, and venom biology has a great potential to engage them with some of the most fundamental questions in biology in an informal and attractive way.

The impact of the research for the broader research community will be secured by disseminating the results through the customary channels of publishing scientific papers, presenting talks at international meetings, and through deposition of the data in the appropriate public databases. The potential impact of the research for the economic and medical competitiveness of the UK will be realized by consulting with two research groups specializing in the development of new drugs from bioactive compounds found in nature, to explore ways to exploit specific pharmacological activities of venom components commercially. The results of this study will also be disseminated to the appropriate people at Defra (Department for Environment, Food and Rural Affairs) to alert them to the value of venoms as a unique natural resource in the UK, and the need to think seriously about their conservation. Finally, the results of the research and the importance of venoms for fundamental and applied science will be communicated to general audiences in several ways. The NHM is an ideal place to do this, with about 5 million visitors per year. First, we will develop and present six presentations on venoms for the popular series Nature Live. These are interactive events in which researchers present their work to Museum visitors, using multimedia and real specimens from the collections. Second, a website on the research will be created and hosted by the Museum's servers for people to look up details. Third, I have made contact with the Head of Biology at St. Paul's secondary school in London (Dr Jon Bennett), and agreed to give several presentations on the research and venoms in general to the pupils. This will expose them to the excitement of science, and how fundamental science and societal benefits can go hand in hand. Fourth, we will arrange a meeting in the Museum with the appropriate people to explore the possibilities of setting up an exhibition on venomous animals in one of the Museum's galleries reserved for temporary (typically several months to a year) exhibits. The Museum's collections are rich in material for this purpose, and should allow a professional and attractive exhibit to be put together.
 
Description The most recent publication I contributed to is necessitating the revision of more than a century old textbook consensus on the structure of the venom system in glycerid polychaetes. What had previously been interpreted as the animals' venom glands are not the main site of venom production. Instead venom is produced in a series of lobe-like structures that have been overlooked for more than a century. This discovery was made in a single species of glycerin worms, analysing the production of a single venom toxin. We are now testing this new insights by looking at several species and the many other venom toxins produced by these worms.
Exploitation Route Our continued discovery of novel venom proteins offers many opportunities to researchers interested in the intersection of fundamental research and applied research, specifically for seeding the development of new pharmaceuticals. We have been approached by other research labs to supply material so that they can study the pharmaceutical activities of the venom toxins.
Sectors Pharmaceuticals and Medical Biotechnology

 
Title Raw data and contigs deposited in GenBank 
Description Raw sequence data and assembled contigs of four transcriptome libraries representing the venom glands of three species of polychaete annelids used in a submitted publication were submitted to GenBank as follows: Glycera tridactyla (body tissue); GART00000000; PID PRJNA234022, Glycera tridactyla; GARU00000000; PID PRJNA234021, Glycera fallax; GARV00000000; PID PRJNA234017, Glycera dibranchiata; GASB00000000; PID PRJNA234012. NCBI/GenBank (https://www.ncbi.nlm.nih.gov) 
Type Of Material Database/Collection of data 
Year Produced 2014 
Provided To Others? No  
Impact No actual impacts realised to date 
 
Description Investigation of polychaete venoms and remipede venoms 
Organisation University of Leipzig
Country Germany 
Sector Academic/University 
PI Contribution We collected specimens for research and contributed to data analyses.
Collaborator Contribution They collected specimens for research and contributed to data analyses.
Impact Two publications, as listed in the relevant section of Researchfish.
Start Year 2012
 
Description Sabbatical project with researcher from GlaxoSmithKline 
Organisation GlaxoSmithKline (GSK)
Country Global 
Sector Private 
PI Contribution We provide research materials and expertise about the animals and venom toxins which allows the collaborator to do their research.
Collaborator Contribution This is in the initial stages, but the GSK scientist will assist us in assaying the bioactivities of venom toxins from venomous polychaetes.
Impact None.
Start Year 2016
 
Description Delivery of temporary exhibition 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I have developed and delivered, as scientific lead, a major temporary exhibition on venom that runs at the Natural History Museum from November 2017 till mid May 2018. The exhibition has attracted lots of positive media attention and reviews, and has so far attracted more than 28,000 visitors. The exhibition highlights in various places that the museum is actively engaged in researching venoms and venomous organisms, and I have personally given tours to many visitors as well as internal colleagues, and been interviewed in print, radio and tv media.
Year(s) Of Engagement Activity 2017,2018
 
Description Departmental talk at Oxford Brooks University 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Talk sparked questions and discussion.

No notable impacts.
Year(s) Of Engagement Activity 2014
 
Description Engagement with general audience during open science night at Natural History Museum 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact The open science event myself and my pre-doctoral research assistant participated in is called Science Uncovered and took place on 30 September 2016. It is the NHM's contribution to the EU funded European Researchers' Night. The event was attended by several thousand people, and we engaged a large number of visitors for 3 hours with specimens and video footage related to venomous animals, highlighting our research. People were amazed that venomous animals are not just spiders, scorpions and snakes, and that the world of venom does a lot for society, including seeding the development of new pharmaceuticals.
Year(s) Of Engagement Activity 2016
 
Description Highlighting of research on website of BBSRC 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact On 29 January and 19 February 2014 the BBSRC's anniversary website of 20 Years of Pioneering Great British Bioscience (http://bbsrc.tumblr.com) featured my lab's research on animal venoms supported by this and related grants.

no actual impacts realised to date
Year(s) Of Engagement Activity 2014
 
Description Interactive public display of venomous animals during NHM event 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact As part of the EU funded Researchers Night I took part in the NHM's Science Uncovered event. I engaged the museum visitors with museum specimens of various venomous organisms. My two postdocs did the same that night in different locations in the museum.

The audience were really enthused.
Year(s) Of Engagement Activity 2014
 
Description Interactive talk about venom research for general audience at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Talk about venom research covered by this and related grants to a general audience at the NHM, followed by showcasing specimens from the Museum's collections.

no actual impacts realised to date
Year(s) Of Engagement Activity 2014
 
Description Invited talk at the Bangor University Student Entomological Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach local
Primary Audience Schools
Results and Impact Invited talk on the research covered by this and related grants to an audience mainly comprising undergraduate students of the University of Bangor and members of the Student Entomological Society

no actual impacts realised to date
Year(s) Of Engagement Activity 2014
 
Description NESCent discussion meeting on animal venoms 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The discussion meeting resulted in the drafting of several manuscripts that summarise current thinking in areas of venomics research, and explore roadmaps for future research in the community.

Broad internal exchange of ideas between participants with very different background, and the drafting of manuscripts together with participants.
Year(s) Of Engagement Activity 2014
 
Description Poster presentation at international scientific meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The presented poster was the centre of scientific discussions.

People were obviously interested in the topic.
Year(s) Of Engagement Activity 2014
 
Description Presentation for general audience at a pub in Crystal Palace, London 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact I presented a talk on venomous animals and my research on them in a pub in Crystal Palace.
Year(s) Of Engagement Activity 2015
 
Description Presentation for general audience at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I presented a talk to an audience of about 50 members of the public on venomous creatures, supported by Powerpoint and specimens from the Museum's collections. This included highlighting my research on centipedes and venomous polychaetes.
Year(s) Of Engagement Activity 2015
 
Description Presentation for general audience at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I presented a talk to an audience of about 50 members of the public on venomous creatures on coral reefs, supported by Powerpoint and specimens from the Museum's collections. This included highlighting my research on venomous polychaetes.
Year(s) Of Engagement Activity 2016
 
Description Presentation for general audience at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I presented a talk to an audience of about 50 members of the public on venomous creatures, supported by Powerpoint and specimens from the Museum's collections. This included highlighting my research on centipedes and venomous polychaetes.
Year(s) Of Engagement Activity 2015
 
Description Presentation for general audience at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Presentation about our research on venomous polychaetes by my postdoc.
Year(s) Of Engagement Activity 2015
 
Description Presentation to A level students at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact I presented a talk with Powerpoint and specimens from the Museum's collections to a group of A level students, highlighting my work as a researcher.
Year(s) Of Engagement Activity 2016
 
Description Public presentation at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I presented a talk to an audience of about 50 members of the public on venomous creatures on coral reefs, supported by Powerpoint and specimens from the Museum's collections. This included highlighting my research on venomous polychaetes.
Year(s) Of Engagement Activity 2015
 
Description Science Uncovered 2017 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact I participated and discussed my ongoing research at the yearly EU funded researchers night, which we call Science Uncovered at the NHM, on the last Friday of September. The activity consisted of showing specimens and discussing the world of venom with the general public who were surprised that the museum is engaged in such exciting scientific research.
Year(s) Of Engagement Activity 2017
 
Description Talk about venom research for general audience at the Natural History Museum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Talk about venom research covered by this and related grants to a general audience at the NHM, illustrated with specimens from the Museum's collections.

Audience members approached me after the event to tell me how interesting they found it.
Year(s) Of Engagement Activity 2013