P2X4 receptor and chronic pain - developing novel pharmacological tools to target P2X4 in the central nervous system

Lead Research Organisation: University of East Anglia
Department Name: Pharmacy

Abstract

Venoms from snakes, spiders, scorpions and marine animals contain a staggering array of peptides and small molecules. Purification of peptide toxins have revolutionised the understanding of the structure and function of ion channels in vertebrates. These highly specific peptide toxins mediate their actions through binding to ion channels and either blocking or enhancing their activity. Typically these peptides are highly potent and selective for individual ion channels, even individual isoforms, and as such these have the potential to be modified to develop new pharmacological tools for research and drug discovery. P2X4 is an important ligand-gated ion channel activated by extracellular ATP and is thought to play a critical role in the development of neuropathic pain. P2X4 regulates Brain Derived Neurotrophic Factor (BDNF) secretion from activated spinal cord microglia and causes alteration of dorsal horn neuronal excitability culminating in pathological pain. This project will identify and characterise a series of peptides isolated from spider venoms acting on the human P2X4 ligand-gated ion channel. The student will learn how to fractionate venoms using reverse phase HPLC to purify active peptide fractions and will characterise their effects using plate reader methods and patch clamp electrophysiology. We will use our novel modulators of P2X4 to investigate the role of this ion channel in microglial responses, in particular the role of P2X4 in secretion of BDNF from microglia using ELISA techniques.

Publications

10 25 50

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Bibic L (2018) Learning to lead. in Science (New York, N.Y.)

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Blasco-Benito S (2019) Therapeutic targeting of HER2-CBR heteromers in HER2-positive breast cancer. in Proceedings of the National Academy of Sciences of the United States of America

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Dhuna K (2019) Ginsenosides Act As Positive Modulators of P2X4 Receptors. in Molecular pharmacology

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Moreno E (2018) Singular Location and Signaling Profile of Adenosine A-Cannabinoid CB Receptor Heteromers in the Dorsal Striatum. in Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011216/1 01/10/2015 30/09/2023
1794654 Studentship BB/M011216/1 01/10/2016 30/09/2019 Lucka Bibic
 
Title Podcasts on various scientific topics and my radio show broadcasted on BBC5Live 
Description As a science communicator and producer/intern for Naked Scientists (University of Cambridge), I have covered topics ranging from medicine to geology, from the environment to rocket science. I have produced over 10 written news stories, conducted interviews with scientists, medical doctors and academics, and produced a total of 15 podcasts, which reached in total to 100K+ downloads. The immediate impact of my work is an improved public understanding of key scientific concepts and an awareness of the advances in various scientific fields. My major outputs are outlined on the Naked Scientists webpage which can be found at the following link: https://www.thenakedscientists.com/users/lucka-bibic As part of the Naked Scientists team, I produced my own 1-hour radio show "Animation: A Reel Deal" which was broadcasted on the BBC (UK) and ABC (Australia) reaching more than 1 million listeners worldwide. This radio show aimed to engage listeners in the science behind animated movies, highlighting how science can help us get from script to screen, from the physics of balancing a giraffe on a tightrope to the researcher putting voice actors in a brain scanner. One of the outcomes of my radio show was an interview with TV celebrity and neuroscientist- Dr. Mayim Bialik of the 'Big Bang Theory' and scientific advisor for Dreamworks studios and a professor in physics - Prof Alejandro Garcia from San Jose State University. Reference to my radio show: https://www.thenakedscientists.com/podcasts/naked-scientists/animation-reel-deal 
Type Of Art Film/Video/Animation 
Year Produced 2016 
Impact My podcasting work has reached a total of 100K+ downloads and my radio show "Animation is the Reel Deal" was broadcasted on the BBC (UK) and ABC (Australia) - it reached more than 1 million listeners worldwide. One outcome of my radio show was an interview with TV celebrity and neuroscientist- Dr. Mayim Bialik of the 'Big Bang Theory' through whom I made useful industry contacts. Furthermore, I co-produced a radio show "Drugs" and lead 3-day science communication workshop for the University of Cambridge, helping international students overcome a language barrier when communicating science. Through attending these workshops, students have the opportunity to learn to communicate science accurately, without using too much jargon. My efforts were recognised by my university (UEA) and I now organize and lead science production workshops for PhD students at UEA, titled as "From Geek to Low-Tech Speak" as part of PPD-credited (professional and personal development) courses which have proven a tremendous success with 90% positive feedback (as reported by PPD coordinator, Dr Sue Jickells) and UEA further contracted me to deliver such workshops annually. REF: (https://www.uea.ac.uk/pharmacy/news-and-events/-/asset_publisher/w7O8j7rUDTtg/blog/coming-soon-science-the-podcast-from-geek-to-low-tech-speak-with-lucka-bibic-?inheritRedirect=false) 
 
Description I am looking at how spider venom toxins might come in handy when treating chronic pain. Our novel screening assays identified the toxins to be uncharacterized small molecules, found in a number of spider species (one manuscript submitted and another in preparation) and this context motivated me to create Bug Off Pain - a VR game that aims to take the pain out of public engagement. Bug Off Pain is an educational game that allows the traditional public engagement process to become more appealing and effective to the general public and students through the use of VR tools.
Exploitation Route Three manuscripts in preparation with Bibic being the first author of the publication and further grant applications to advance these findings.
Sectors Education,Pharmaceuticals and Medical Biotechnology

 
Description I am looking at how spider venom toxins might come in handy when treating chronic pain. So far, our novel screening assays identified the toxins to be uncharacterized small molecules, found in a number of spider species and this context motivated me to create Bug Off Pain - a VR game that aims to take the pain out of public engagement. Bug Off Pain is an educational game that allows the traditional public engagement process to become more appealing and effective to the general public and students through the use of VR tools. As a player, you must figure out which spider venom is capable of shutting down the pain until the game ends with "pain over". Interestingly, our study measuring the effectiveness of Bug Off Pain revealed that VR is a better learning method then video. The game is now freely downloadable via our website www.bugoffpain.com. One potential drawback of using VR is the accessibility of the VR equipment to the general public. For example, top-of-the-range systems such as the Oculus Rift can be prohibitively expensive. However, the Google Cardboard system, which uses ones smartphone and a cardboard phone holder at the expense of some immersive experience, provides an accessible way of implementing VR since the majority of people now use the smartphones. Since we strived to get the best of the both worlds, we launched Bug Off Pain on both platforms. An additional challenge was that motion sickness can be a problem with VR systems due to the small delay in the user making a movement and the screen showing that movement. Therefore it is usually recommended not to play VR games for more than 20 minutes at a time. We therefore aimed for a user experience between 7-10 minutes long and our later evaluation reported zero VR-related sickness during gameplay. Once we started evaluating the game in terms of 1) the general public's opinion about Bug Off Pain, and 2) VR-based learning using Bug Off Pain among high school students, we had to utilize educational research skills and team up with the educators at University of East Anglia to design effective questionnaires and forms that would effectively assist us in these evaluations. Here, we had to consider the design and development of self-administered surveys to assessing student learning, employ them and, in turn, using that information to improve teaching. Bug Off Pain brought together a wide range of partners from academia and education authorities, research active schools, science communicators, students and software developers with an interest in unlocking not only the massive opportunity already exposed, but in implementing long-term and sustainable quality teaching practices in schools. Launching the game on the Norwich Science Festival helped us to reach out to more people and offer immersive educational content through VR. Crucially, VR offers insight in a way you simply can't get in a classroom through traditional methods, as this immersive experience fosters learning retention that beats a text book or PowerPoint presentations. After Bug Off Pain's successful launch at the Norwich Science Festival in 2017, the game has also been very favorably received at international academic conferences (notably at the ACS National meeting 2018 in Boston), online and other events, such as "Cambridge Science Festival", "Norwich Gaming Festival" and "UEA Innovation Showcase" in 2018. Feedback from the users online or attending these events has been universally positive. Specifically, by using pre - and post-tests, our evaluation showed the effectiveness of the instructional role of the game in promoting learning about chronic pain and the biochemistry of venoms via a virtual reality-medium. Moreover, by using a 10 point Likert-type scale, we show that the game is fun, dynamic, easy to play, has an attractive easy to understand interface and relevant content. Interestingly, our data suggest that extending Bug Off Pain to other STEM-relevant subjects might be educationally beneficial as well. Together, our findings reveal that virtual reality games, such as Bug Off Pain, allow the traditional public engagement process to become more appealing and effective to the general public and students. Since we believe that our findings would be of significant interest to the other educators and researchers that would like to employ similar ideas to their own field, Bibic submitted our work as a Technology Report (Manuscript ID: ed-2018-009052) to ACS Chemical Education Research journal which is now under minor revision. Moreover, our results have clear implication on how to design and implement virtual reality games in chemistry education and offer insights into the implications for the broader teaching practice. The last, but not the least, Bug Off Pain directly addressed the BBSRC strategic goals of lifting productivity and impact on society with creating an innovative platform called Bug Off Pain.
First Year Of Impact 2017
Sector Education,Pharmaceuticals and Medical Biotechnology
Impact Types Cultural,Societal

 
Description Biochemical Society Travel Grant
Amount £550 (GBP)
Organisation Biochemical Society 
Sector Learned Society
Country United Kingdom
Start 08/2018 
End 08/2018
 
Description Royal Society of Chemistry Travel Grant
Amount £750 (GBP)
Organisation Royal Society of Chemistry 
Sector Learned Society
Country United Kingdom
Start 08/2018 
End 08/2018
 
Title Bug Off Pain: an educational virtual reality game on spider venoms and chronic pain for public engagement 
Description Bug Off Pain is an immersive, interactive and educational virtual reality (VR) game that increases scientific literacy about chronic pain and spider venoms among the general public and high school students. Here, VR was shown to be an innovative and fun approach to learning and public engagement in biochemistry. Bug Off Pain places the viewer inside the brain and shows the molecular system that allows people to sense pain. After securing three (learning) points via the multimedia-based clips, this experience translates to the interactive game. Here, a player has to choose a venom that shuts down the pain until that results in "pain over". Bug Off Pain can be played (free of charge) on two different VR platforms; Oculus Rift and Android devices as a downloadable software application. 
Type Of Technology Webtool/Application 
Year Produced 2017 
Open Source License? Yes  
Impact After the initial launch, "Bug Off Pain" attracted the attention of more than 500 people attending the festival and has been favorably received at the others academic conferences and events such as "Norwich Gaming Festival 2018" and "UEA Barclay's Showcase Event" in 2018. We've also undertaken two crucial evalutations for the game: 1) evaluation of general public opinions about Bug Off Pain" and 2) evaluation of VR-based learning by use of Bug Off Pain among high school students. The manuscript is currently in preparation for the publication in a scientific journal. The current download score on Bug Off Pain is: 4315 (checked on the 18/09/2018). WEB REFERENCE: http://bugofpain.s3-website.eu-west-2.amazonaws.com/ 
URL https://www.uea.ac.uk/pharmacy/research/bug-off-pain
 
Description Interview with Scientists: Lucka Bibic 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Invited to participate in HelloBio "Interviews with Scientists" series which reached 3.4k+ views.
Year(s) Of Engagement Activity 2017
URL https://www.hellobio.com/blog/interviews-with-scientists-lucka-bibic.html
 
Description Making ice-cream out of the liquid nitrogen 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact As part of the Norwich Science Festival 2017 launching event, I and my colleague (Oliver Cartwright) were asked to show and explain the chemistry behind making ice-cream out of the liquid nitrogen. Following this event, we were asked to come to various schools around Norfolk (UK) and deliver the same workshops for kids aged 8-12.
Year(s) Of Engagement Activity 2017
URL https://twitter.com/LuckaBibic/status/921074925481414656
 
Description Norwich Science Festival 2017 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact We provided engaging content with specific subject matter expertise about spider venoms that will provide essential insights on scientific trends in accordance to the best practices to the audience at the Norwich Science Festival 2017. This project aimed to target students and smash stereotypes of scientists, improve understanding of how science works, give a more nuanced view of science, broaden understanding of scientific disciplines and making science more relevant and obtainable for students. "No More Ouch: Brewing better painkillers from the spider venoms" was a science engagement event, launched at the Norwich Science Festival in 2017, and had reported being an engaging way of learning about spiders, their webs, fangs and its venom and can be used for public outreach and education. These workshops consisted of two main events: 1) a virtual reality (VR) game called "Bug Off Pain" as an innovative approach to public engagement and education. This immersive and educational VR experience which places the viewer inside the brains and shows them first-hand the molecular system in our brains that allows us to sense the pain. And 2) creating spider webs from hula hoops and colored wool. Once a hula hoop web is created, kids glued colored spiders on it and put glitter on their fangs as the venom. This activity entertained and educated them in equal measures and they also became familiar with the spider's body parts and their functions. (https://twitter.com/Stokes_Lab/status/923943974359977984) This was featured on UEA Live TV and broadcasted via Facebook platform (https://www.facebook.com/ueaofficial/videos/discover-more-about-the-spooktacular-powers-of-spider-venom-and-the-vr-game-bug-/1590537734337169/ ) and reached 4,3k people.
Year(s) Of Engagement Activity 2017
 
Description Video clip about how to make icre-cream out of liquid nitrogen 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact http://www.uea.ac.uk/pharmacy/news-and-events/-/asset_publisher/w7O8j7rUDTtg/blog/the-brilliant-club-tutor-oliver-cartwright-chills-out-at-colman-junior-school?inheritRedirect=false
Year(s) Of Engagement Activity 2017
URL https://www.youtube.com/watch?v=4Zo3P7x_jlU