Elucidate exosome transfer between cells and determine the role of caspases in the regulation of exosome assembly and secretion
Lead Research Organisation:
University of Oxford
Department Name: Interdisciplinary Bioscience DTP
Abstract
Establishing a novel biological model to elucidate exosome transfer between cells and determine the role of caspases in the regulation of exosome assembly and secretion under physiological conditions.
Exosomes are a subtype of extracellular vesicles that efficiently transport proteins, lipids and RNA, between cells. Although their instrumental role during cell communication is broadly accepted, the basic molecular mechanisms responsible for their loading and cellular release remain largely unknown. We are developing a Cre-LoxP reporter system suitable for investigating exosome loading and transfer between Drosophila S2 cells and fly tissues. Preliminary results using this system in cell culture based assays suggest that Cre mRNAs can be potentially packaged within extracellular vesicles and transferred between cells. Interestingly, specific exosome-loading and subsequent transfer of mRNAs can be virtually enhanced, by adding a 25-nucleotide extracellular vesicle 'zipcode'. Our tool is currently in the process of adaptation within complex Drosophila tissues, aiming to investigate further the biology of exosome-mediated cell communication. In this regard, we will focus our scientific efforts on uncovering the molecular features of mRNAs that facilitate the loading within exosomes using RNA-Seq analysis. In addition, we will investigate the involvement of the apoptotic factors known as caspases in exosome release. Finally, we aim to exploit this knowledge to change cellular properties remotely, by using exosomes loaded with customised cargos. Our project holds the potential to improve our fundamental understanding of exosome loading/transfer, whilst paving the way to the development of biological tools to manipulate intercellular communication via exosomes.
BBSRC Priorities Covered
The replacement, refinement and reduction (3Rs) in research using animals - Adapting the Cre-LoxP system for studying exosome loading and transfer to Drosophila cell culture and in vivo models, reducing the need to utilise a comparable system established in mice.
Technology development for the biosciences - Drosophila cell based and in vivo models to investigate the molecular mechanisms of exosome loading and their potential as molecule carriers, providing a novel resource to the EV and biological research communities.
World-class bioscience - Using a collaborative approach and varied biological/bioinformatic techniques to answer fundamental biological questions.
Bioscience for health - Exosomes have been implicated in a number of diseases and thus, understanding their fundamental biology improves our understanding of their role in disease. Our work paves the way to harnessing the ability to utilise exosomes for the loading, release and directed transfer of therapeutic molecules for drug delivery.
WUB, ENWW
Exosomes are a subtype of extracellular vesicles that efficiently transport proteins, lipids and RNA, between cells. Although their instrumental role during cell communication is broadly accepted, the basic molecular mechanisms responsible for their loading and cellular release remain largely unknown. We are developing a Cre-LoxP reporter system suitable for investigating exosome loading and transfer between Drosophila S2 cells and fly tissues. Preliminary results using this system in cell culture based assays suggest that Cre mRNAs can be potentially packaged within extracellular vesicles and transferred between cells. Interestingly, specific exosome-loading and subsequent transfer of mRNAs can be virtually enhanced, by adding a 25-nucleotide extracellular vesicle 'zipcode'. Our tool is currently in the process of adaptation within complex Drosophila tissues, aiming to investigate further the biology of exosome-mediated cell communication. In this regard, we will focus our scientific efforts on uncovering the molecular features of mRNAs that facilitate the loading within exosomes using RNA-Seq analysis. In addition, we will investigate the involvement of the apoptotic factors known as caspases in exosome release. Finally, we aim to exploit this knowledge to change cellular properties remotely, by using exosomes loaded with customised cargos. Our project holds the potential to improve our fundamental understanding of exosome loading/transfer, whilst paving the way to the development of biological tools to manipulate intercellular communication via exosomes.
BBSRC Priorities Covered
The replacement, refinement and reduction (3Rs) in research using animals - Adapting the Cre-LoxP system for studying exosome loading and transfer to Drosophila cell culture and in vivo models, reducing the need to utilise a comparable system established in mice.
Technology development for the biosciences - Drosophila cell based and in vivo models to investigate the molecular mechanisms of exosome loading and their potential as molecule carriers, providing a novel resource to the EV and biological research communities.
World-class bioscience - Using a collaborative approach and varied biological/bioinformatic techniques to answer fundamental biological questions.
Bioscience for health - Exosomes have been implicated in a number of diseases and thus, understanding their fundamental biology improves our understanding of their role in disease. Our work paves the way to harnessing the ability to utilise exosomes for the loading, release and directed transfer of therapeutic molecules for drug delivery.
WUB, ENWW
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011224/1 | 01/10/2015 | 31/03/2024 | |||
| 1757829 | Studentship | BB/M011224/1 | 01/10/2016 | 31/03/2021 |
| Title | Science in Ten Podcast |
| Description | Science is discovering things that have never been discovered before. It is one of the most exciting fields. There is so much science research going on out there, it is hard to keep track! Claire Hill, a PhD student at Oxford University, takes a new topic each month - summarises the science, outlines the debates and excites us about the future. |
| Type Of Art | Artefact (including digital) |
| Year Produced | 2019 |
| Impact | Advancing skills in science communication. |
| URL | https://podcasts.ox.ac.uk/series/science-ten |
| Description | Collaboration with Dr Alberto Baena |
| Organisation | University of Oxford |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We have provided our expertise in EVs to establish a new collaboration to investigate EV transfer and stress response using the model organism Drosophila Melanogaster. |
| Collaborator Contribution | We provided intellectual input and help to supervise two PhD students working directly on this collaboration. |
| Impact | The students are in their second year and have produced excellent preliminary data which are being prepared for a manuscript |
| Start Year | 2017 |
| Description | Articles in Oxford 'Phenotype' Magazine |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | A number of my articles have been published in the Oxford University 'Phenotype' magazine. "The biochemistry of Fragile X Syndrome (FXS)" Michaelmas Term 2017 "The mystery of planarian regeneration" Hilary Term Issue 2017 "Caspases and their roles beyond death" with Dr. A. Baena Lopez Hilary Term Isssue 2019 I wanted to get involved in writing articles to improve my ability to discuss science topics to a non-specialist audience. |
| Year(s) Of Engagement Activity | 2017,2019 |
| URL | https://issuu.com/phenotypejournal/docs/mt17-1/20 |
| Description | Co-organiser and speaker at the Balliol College Life Science Symposium 2018 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | I was the co-organiser of the Balliol College Life Science Symposium, an annual event at my Oxford college to encourage PhD students to share their research in fun and exciting ways to other life science students and the entire Balliol postgraduate community. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Curator and host of panel event at Northern Ireland Science Festival 2019 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | As part of my PhD internship with Northern Ireland Science Festival I designed a panel event called 'Diagnose me Please, Treat my Disease'. I invited three speakers, Prof. Dave Carter, Prof. Denise Fitzgerald and Prof. Patricia Maguire to discuss how their cutting-edge research was paving the way to new diagnostics and therapeutics. I invited speakers who had overlapping areas of interest; feedback indicated that this helped audience understanding. Feedback was also received that the use of a casual café/pub venue (Black Box, Belfast) was less intimidating and encouraged attendance by non-scientists. All speakers did a wonderful job explaining their work in an accessible way. As the event host, I focused on making the audience feel comfortable, encouraging them to get involved and leave feeling inspired. One visitor requested more of these events throughout the year. The event was sold out and reached approximately 40 people. This event may have also sparked collaborations between speakers. |
| Year(s) Of Engagement Activity | 2019 |
| URL | https://www.nisciencefestival.com/event.php?e=89 |
| Description | Design and delivery of an interactive activity to educate the general public and children about how extracellular vesicles can be used in disease diagnosis. |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | As part of my PhD internship with Northern Ireland Science Festival (NISF) I designed an activity, called 'Disease Detectives', to educate children aged 4-8 about the use of extracellular vesicles (EVs) to diagnose disease. I delivered the activity alongside my friend, Patrick Martin, as part of the NISF Roadshow; a touring, four-day event across NI, visiting Newry, Enniskillen, Dungannon and Ballymena. This event allowed families from rural areas of NI to visit NISF; engaging a wider section of the community. It is estimated that the Roadshow had 2,400 visitors in total. Some feedback received included: • 'Really good way to put it across.' • 'Age appropriate.' • 'You should make this into a book to explain disease simply and well, when it can be a scary subject.' • 'Well explained, both for kids and for Joe Bloggs.' • 'Gets kids interested.' • 'Explained really well.' • Proof that our explanations were well understood by children was when we heard comments such as 'the bad guys are sending me a message' whilst pointing at the ball pit. • One child (around 3 years old), ran to his mum after receiving his sticker inside his EV saying, 'I got a message, I got a message.' • Some children came back to help us explain the stand to their siblings. • Many adults were excited to hear about this area of research and were asking questions to find out more. • Some children said it was their favourite, or one of their favourite, Roadshow activities. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Flash Talk at UK Society for Extracellular Vesicles (UKEV) Forum |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Other audiences |
| Results and Impact | Presentation describing my project, "The identification of extracellular vesicle enriched mRNA features which facilitate long-range mRNA transfer in Drosophila" to the scientific community focused on extracellular vesicles. |
| Year(s) Of Engagement Activity | 2020 |
| Description | Lightning Talk and Poster Presentation, UK Society for Extracellular Vesicles Meeting, Francis Crick Institute. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Other audiences |
| Results and Impact | Presenting our recent findings via a Poster presentation and lightning talk entitled "Biological principles of mRNA loading into extracellular vesicles uncovered by a novel bioinformatic pipeline and in vivo approaches." Presentations were made at the UK Society for Extracellular Vesicles Annual Meeting in the Francis Crick Institute, London. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Poster presentation, Annual Oxford Developmental Biology Symposium, DPAG, Oxford. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Other audiences |
| Results and Impact | Poster presentation entitled 'Biological principles of mRNA loading into extracellular vesicles uncovered by a novel bioinformatic pipeline and in vivo approaches' given at the Annual Oxford Developmental Biology Symposium, DPAG, Oxford. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Science Fair outreach at Oxford Science Festival |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | I was part of a team who presented the work of the Extracellular Vesicles Brookes University lab to the general public. This took place during the Science Bazaar 2017 in the Oxford Town Hall. The general public, particularly children with their parents, were engaged with our hands on stall and left having learnt about extracellular vesicles, cell communication and disease. |
| Year(s) Of Engagement Activity | 2017 |
| Description | Speaker at an Oxford Pathways taster day |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Schools |
| Results and Impact | I described my research to school children (GCSE to A Level age) at an Oxford Pathways event. Oxford Pathways is designed to provide information and insight into the different courses available at Oxford, and provide guidance on higher education in general, to students and teachers from non-selective state schools with little history of pupils applying and attending Oxford University. This was a great opportunity for me to explain my research in an accessible and engaging way. My presentation sparked questions about studying science in general and more specific questions about the research I had discussed. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Speaker at the 2018 Oxosome Meeting |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | Speaker at Oxosome meeting with talk entitled "Novel in vivo tools for investigating EV-mediated cell communication." This was the first opportunity for me to present my work and it sparked collaboration with another research group at Oxford University. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Talk at International Society for Extracellular Vesicles (ISEV) Annual Meeting |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | A virtual talk entitled 'A novel in vivo Drosophila model of vesicle-mediated intercellular communication', describing my research to the international scientific community focused on extracellular vesicles. |
| Year(s) Of Engagement Activity | 2020 |
| URL | https://www.tandfonline.com/doi/full/10.1080/20013078.2020.1784511 |