In this Repurposing TIN interview as part of the Early Career Innovators series, acknowledging the amazing translational work being done by early career and non-tenured researchers within the UCL Therapeutic Innovation Networks (TINs), Dr Melissa Rayner highlights her Repurposing TIN Pilot Data Fund awarded project, establishing a relationship between a drug’s affinity for PPARγ and nerve regeneration.

What is the title of your project and what does it involve?

The title of the project is ‘Investigating the correlation between PPARγ affinity and nerve regeneration’. Previous studies I conducted demonstrated the beneficial effects of the PPARγ agonist, ibuprofen, on regeneration and functional recovery following a peripheral nerve injury. However, in vitro data has shown that PPARγ agonists with a higher affinity correlate to greater regeneration. Therefore this study will allow us to determine whether a corresponding correlation occurs in vivo. Once this is established the optimal drug could be taken forward and developed to be delivered locally to an injured nerve through a biomaterial or injection.

What is the motivation behind your project/therapeutic?

Peripheral nerve injuries have a high prevalence and can be debilitating, resulting in life‐long loss or disturbance in function, which compromises quality of life for patients. Current therapies use microsurgical approaches to repair the nerve but there is the potential for enhancing recovery through other therapies. Following an injury, a repaired nerve can regenerate and grow back to the target organ (e.g. muscle or skin) to restore function, however, outcomes are generally poor with incomplete restoration of function. This is due to regeneration being remarkably slow, ~1mm/day. The problem is that there are no current treatments available to increase this regeneration rate. The research we have done has identified drugs and a local delivery platform that could provide a therapy for this clear clinical need.

Why did you want to apply to the Repurposing TIN Pilot Data Fund?

Following an initial experiment that I conducted during my PhD I had found an interesting correlation between a drug’s affinity for PPARγ and its capacity to increase nerve regeneration. I wanted to confirm whether this correlation is reproduced in vivo but unfortunately I had no funding to conduct the study. I applied for the TIN pilot fund so I was able to take this work forward and move it closer to the clinic. In addition, the TIN pilot fund gave me an opportunity to apply for a grant as the lead applicant which is a stepping stone in my development towards my goal of building an independent research career.

The UCL TINs are hoping to run another round of Pilot Data Funds in the summer. Subscribe to the TINs newsletter to keep updated.

How did you find the process for the TIN Pilot Data Fund? What did you learn?

The application process gave me the opportunity to write my first grant application as a lead applicant. Furthermore, the application process involved a Dragon’s Den event where we had to pitch our project/therapeutic. This was a new experience and allowed me to develop a lot of new skills. I had the great opportunity to attend an ACCELERATE pitching coaching session and receive one-to-one feedback on my presentation style.

What do you hope to achieve in the 6 months duration of your project?

I hope to be able to establish a relationship between a drug’s affinity for PPARγ and nerve regeneration in vivo which will enable us to identify a leading repurposed drug for use in peripheral nerve injury. I can then further develop local controlled drug treatments with a more potent PPARγ agonist which may be more commercially competitive than ibuprofen. I hope this will provide positive data which can be used in subsequent bids for funding which will allow me to move this work closer to translation into the clinic.

About Dr Melissa Rayner

Dr Melissa Rayner qualified as a pharmacist in 2014 before starting a PhD at the Centre for Doctoral training in Advanced Therapeutics and Nanomedicines at UCL School of Pharmacy. Her PhD was a multi-disciplinary project combining tissue engineering and drug development to improve regeneration and functional recovery following peripheral nerve damage. During this time she also worked for the UCL spin-out company, Glialign, to develop a stem cell based tissue engineered product to repair peripheral nerves.

Melissa is currently a postdoctoral research fellow working in the UCL Institute of Prion Disease on a project funded by the Department of Health. The project involves developing a cell based assay that could be translated to the clinic as a tool to diagnose Creutzfeldt-Jakob disease. Through collaborations with UCL School of Pharmacy she still continues her research on regenerative medicine for peripheral nerve injury, including work on the development of local drug therapies to improve regeneration.

In the first Devices & Diagnostics TIN interview as part of the Early Career Innovators series, acknowledging the amazing translational work being done by early career researchers within the UCL Therapeutic Innovation Networks (TINs), Dr Hayley Pye highlights her Devices & Diagnostics TIN (co-lead by the UCL Institute of Healthcare Engineering’s Translational & Industry Delivery Group) Pilot Data Fund awarded project, “Personalising pathology for prostate cancer, a unique opportunity at UCL/UCLH”.

What is the title of your project and what does it involve?

I was recently awarded money from UCL TRO’s Therapeutic Innovations Network, specifically the Devices & Diagnostics TIN, to carry out some proof on concept work predicting molecular pathways from diagnostic histology images. The project is called: “Personalising pathology for prostate cancer, a unique opportunity at UCL/UCLH.”

Prostate cancer is the 2nd most common cause of cancer death in males in the UK with over 11,000 men dying each year. Around 350 people are diagnosed with prostate cancer every day. In the UK there are no molecular profiling tests recommended for risk profiling or treatment selection. Despite high quality research in this area, molecular tests to diagnose only the more aggressive cancers that require treatment are largely unadopted. Our device will provide a completely novel way to implement personalised diagnosis and pave the way for the implementation of these tests.

Every cancer diagnosis requires the analysis of a histopathology image, this image (right) is produced from staining a human tissue sample with two dyes (hematoxylin and eosin).

I want to apply machine learning techniques to these whole-slide-images (WSI) and predict for clinically actionable genetic mutations or pathway activation in prostate cancer. This would promote personalised therapy from the point of diagnosis.

Our network provides a unique environment to test this in clinical hands and in real life clinical data sets that are relevant to the target population.

What is the motivation behind your project/therapeutic?

Traditional ‘one size fits all’ treatments for prostate cancer can cause a lot of off target urinary, sexual and bowel symptoms, and overtreatment is common. Our device ultimately will be pathway to implement augmented filters that would overlay an individual’s standard of care diagnostic whole-slide-images (WSI) and inform the patient if they could be spared treatment and/or if a modern targeted treatment could work. This device would help introduce molecular profiling at diagnosis (not yet standard of care) and therapy choice based on the individual cancer’s molecular characteristics.

The diversity in molecular characteristics present in prostate cancer means simple molecular signatures often either struggle to predict treatment sensitivity accurately, or are not present in sufficient numbers of patients. In depth genetic and transcriptomic sequencing can provide comprehensive information about individual tumours but it is expensive, requires the destruction of tissue samples, and the time to result can be long. The translation of in depth genetic and transcriptomic sequencing tests into clinical implementation is challenging due to issues of scale, as well as financial and infrastructural limitations in many centres. The ubiquity of the ‘standard of care’ histopathology slides alongside our device would allow for quick and cheap pre-screening of patients and without any additional tissue.

Why did you want to apply to the Devices & Diagnostics TIN Pilot Data Fund?

Since I am a postdoctoral research associate, the opportunities to raise money and lead your own research project are rare and competitive. However they are also essential if you want to continue a career in academic research. These opportunities are especially challenging as it all has to be done on top of an already busy and challenging day job as a research associate. The TIN Pilot Data Fund is perfectly sized and suited for an opportunity to carry out some independent research within my current role at UCL. In addition, the amount of support and guidance is phenomenal, ideal for someone like myself trying to make that first step into independence.

Learn more about TIN opportunities for researchers

How did you find the process for the TIN Pilot Data Fund? What did you learn?

I felt very supported and encouraged throughout the process. I am still learning and hoping to take advance of all that ACCELERATE can provide, they are very approachable and all the content so far has been appropriate and all very reasonable and relevant. I have attended the following courses: ‘Mentoring for Medical Innovation’, ‘Grant Writing for Translational Research’ and a ‘Pitching Skills Workshop’ as well as applying for a ‘Dragon’s Den’ style grant from the Devices & Diagnostics Therapeutic Innovations Network.

Register for upcoming ACCELERATE events

What do you hope to achieve in the 6 months duration of your project?

Funds from this grant will help to progress a proof of concept computer algorithm through to testing on real-world diagnostic data. Work will involve curating and cleaning the cohort(s) for this type of work, building a strong hypothesis on which to build a model, generating molecular label data for the test cohort, designing suitable approaches for the data pipeline, and finally applying at least one algorithm to real-world diagnostic data. Hopefully I will then be in a strong position to apply for further funding, leveraged by this work, to both access a larger clinically relevant biobank, expand the algorithms to other molecular signatures and embed our tools into a commercial platform already embedded for NHS-wide deployment.

What are your next steps from now?

The most difficult and time-consuming task will be generating reliable and high-quality molecular label data. So this is my main focus at the moment.

About Dr Hayley Pye

Dr Hayley Pye is a Postdoctoral Research Associate based in the Division of Surgery at UCL. She is currently working on a clinical trial evaluating new blood and urine biomarkers in combination with MRI imaging for improved prostate cancer diagnosis and prognosis. She also has experience in the early-stage development of antibody-drug conjugates (ADC) that can be activated by light (photodynamic) sound (sonodynamic) or carrying chemotherapeutic drugs for cancer.

She is driven to better understand the way we currently diagnose and choose modern therapeutics for cancer in the clinic and wants to disrupt the stratification by ‘organ of origin’ and the research silos we currently work in.

In the first Small Molecules TIN interview as part of the Early Career Innovators series, acknowledging the amazing translational work being done by early career researchers within the UCL Therapeutic Innovation Networks (TINs), Clara Gathmann highlights her Small Molecules TIN Pilot Data Fund awarded project, “Screening a DNA encoded library on GEF-H1 for drugs targeting ocular diseases”.

What is the title of your project and what does it involve?

The title of my project is ‘’Screening a DNA encoded library on GEF-H1 for drugs targeting ocular diseases’’. GEF-H1 is a protein which our group has identified as a potential target for fibrotic and inflammatory ocular disorders. We have started to develop small molecules against GEF-H1, using computational and medicinal chemistry drug discovery techniques. However, our molecules are not very potent yet and we haven’t tried using high throughput screening.

DNA encoded libraries (DELs) contain billions of molecules in a single tube and can be screened on a protein within days. This technology is made possible by DNA tags attached to each molecule that encode their structures. As you may know, DNA can be amplified with PCR and sequenced. This enables a read-out of the structures of the most active molecules from picogram quantities in the tube. Hence, only one small tube is needed instead of thousands of 96-well plates. We decided to use the open source DEL library from WuXi and subject it to GEF-H1. With this, we hope to discover new molecules that bind GEF-H1 within a few weeks, giving a real kick to our drug discovery plans!

What is the motivation behind your project/therapeutic?

Preventable ocular disorders are still a major cause for vision loss. In addition to the high impact on human lives, sight loss is also a real economic issue for our societies. Even quite common ocular disorders like uveitis and retinopathies can cause vision loss if left untreated. Unfortunately, current treatments for inflammatory and fibrotic eye disorders either involve invasive surgeries or the heavy use of drugs like corticosteroids and anti-proliferative agents. These interventions are first of all not necessarily successful and have several side effects, including even vision loss!

As for many diseases, it all starts with a good drug target (in other words, a protein to inhibit). By finding potent GEF-H1 inhibitors, we hope first of all to produce useful clinical candidates that can prevent inflammatory and fibrotic damages done to the eye. But also importantly, we would show for the first time that GEF-H1 is a suitable protein to inhibit, paving the path for a new class of biological targets and providing a proof of concept for future drug discovery projects. I think this is truly motivating, to not only contribute to a cause such as a particular disorder, but also to a whole scientific field which might help completely unrelated disease classes.

Why did you want to apply to the Small Molecules TIN Pilot Data Fund?

DNA encoded libraries are an emerging technology that have gained a lot of attention lately. More and more data is published on the construction of such libraries, with some successful examples taken to clinical trials. However, the biggest libraries are often in-house libraries of pharmaceutical companies or offered by specialised companies to pharma giants. This means that drug discovery groups in academia don’t have easy access to these libraries for cost reasons, unless a collaboration is put in place. When we heard that WuXi launched this open source DNA encoded library for academia, it sounded like a huge opportunity to bring that technology into UCL and our department.

This technology can lead very quickly to positive results but would simply not have been possible to follow up on hits without the funds from the Small molecules TIN. The TIN pilot data fund seemed to be adapted due to the short-term character of the project and we hoped that such an unusual idea could awaken interest. In addition to this, applying for this kind of fund seemed like the perfect opportunity for me to learn about grant writing and how to fund research in general.

Learn more about the Therapeutic Innovation Networks and join a TIN

How did you find the process for the TIN Pilot Data Fund? What did you learn?

Overall, super exciting. While making the written application, I realised how important it is to connect our (sometimes crazy) scientific ideas to real life goals. For the first time in my life, I had to define the purpose for what I am doing in such details. In some disciplines like the ones involving clinical research, it might be easier to relate to the patients, but when you are in a lab synthesising molecules, sometimes you just loose that connection. Applying for the fund made me realise that the translational aspect of research even at its early point is really important.

Then, there was the pitching, which felt a bit like preparing for a TV show! We candidates had the chance to participate to an ACCELERATE workshop on pitching, probably the most important part of the application process. My pitch before and after that session was transformed thanks to the honest comments I received. I learnt to shift completely the initial ‘science nerdy’ focus of the talk to ‘why you should fund my project’. In summary, I learnt to tell why my project is going to make a change, and how I am going to achieve my goals in time.

What do you hope to achieve in the 6 months duration of your project?

The goal for this project is mainly to generate molecules that can be useful biological tools or clinical candidates which target GEF-H1 in ocular diseases. We want to generate accurate binding data on the hit molecules to prepare them for in vivo testing. If the screen generates potent molecules that bind GEF-H1 tightly, this will enable us to irrevocably confirm that GEF-H1 inhibition is beneficial for those diseases, fast-tracking us to clinical testing.

What are your next steps from now?

In those six months, I will first prepare materials for the screen (immobilise the proteins on beads for example), to then subject the protein to the DEL screen. After the screen is done, WuXi will process our samples, amplify the DNA tags and perform a statistical analysis on these to reveal the structures of the binders. We will then order the most potent binders for resynthesis on a milligram scale and validate them using biological assays. We typically perform biophysical assays like surface plasmon resonance (SPR) and cellular assays that model for example inflammation.

About Clara Gathmann

Clara Gathmann works between the UCL Institute of Ophthalmology and the Wolfson Institute for Biomedical Research. She is working in the groups of Prof. Balda/Matter and Dr.Chan/Prof. Selwood, focussing on the discovery of small molecules as drug candidates for common ocular diseases. She started on a Moorfields Eye Charity funded project in October 2019 involving the design, synthesis and biological assessment of molecules inhibit the GEF-H1/RhoA interaction.

In the next interview as part of the Early Career Innovators series, acknowledging the amazing translational work being done by early career researchers within the UCL Therapeutic Innovation Networks, Dr Athina Dritsoula and Dr Carlotta Camilli highlight their joint Biologics TIN Pilot Data Fund awarded project focusing on the effect of LRG1 blockade in pancreatic cancer.

How did this joint project come about?

CC: Athina and I both arrived at the Institute of Ophthalmology to do our post-docs where, weirdly enough, we don’t do eye-related research but explore vessel behaviour using ex-vivo models of angiogenesis and in vivo models of solid tumours.

What is the title of your project and what does it involve?

AD: The interest of our lab is focused on the LRG1 protein, which is involved in pathological angiogenesis, but we don’t know much about its normal function. Based on our research, we believe that blocking the function of the LRG1 protein by using a specific monoclonal antibody that we have developed in the lab will be beneficial in conditions with abnormal angiogenesis like cancer. In fact, LRG1 is upregulated in pancreatic cancer, which is a type of cancer with minimal survival that remains untreated. So, Carlotta and I designed this project to study the effect of LRG1 blockade in pancreatic cancer.

What is the motivation behind your project/therapeutic?

CC: Pancreatic cancer is a leading cause of deaths from cancer that kills about half a million people worldwide each year. The current standard of care involves combination cytotoxic chemotherapy, which often fails due to the complex tumour microenvironment. So, there is a great need for developing novel therapeutic strategies that will target new molecules and pathways, and we believe that our anti-LRG1 antibody could be a great novel therapeutic candidate.

Why did you want to apply to the Biologics TIN Pilot Data Fund?

AD: We both thought that the Biologics TIN Programme is a great opportunity to get enough funding to support a short 6-month project that would allow us to a) test our hypothesis and b) generate pilot data to design a bigger project in future if (a) proves right.

Join the Biologics TIN

What do you hope to achieve in the 6 months duration of your project?

CC: Well, we hope that we will manage to prove that our hypothesis is correct, and COVID-19 situation allowing, generate data to apply for more funding and get this project further. Our -very ambitious – aim is to get our antibody into clinical trials for pancreatic cancer in a few years’ time!

What are your next steps from now?

AD: Hope that our orders will be delivered on time and that our experiments will work as planned! We need to complete the project before a second lock down crushes.

Do you have any top-tips for applicants currently going through the application process for the other TIN Pilot Data Funds?

AD: The 10 minutes Dragons’ Den round felt much longer than it was! Be well prepared for all types of questions, and maybe have a mock interview with your PI, if possible.

CC: Spend enough time to prepare the slide presentation. It might seem easy but it’s not, as it needs to be very concise and straight forward!

About Dr Athina Dritsoula

Dr Dritsoula studied an undergraduate degree in Molecular Biology and Genetics in Greece over a decade ago before arriving in the UK for a Master’s and PhD, and UCL has been home to Dr Dritsoula since. Although human genetics was Dr Dritsoula’s first love, Dr Dritsoula quickly found her “forte” in vascular biology – studying the biology of big human vessels during a PhD, and then smaller vessels stability and angiogenesis during post-doc.

About Dr Carlotta Camilli

After completing a Master’s in Medical Biotechnology, Dr Camilli left Italy to start a PhD at UCL focusing on the use of vascular progenitors for the development of a bioengineered muscle.

However, Dr Camilli’s broad interest in translational medicine pushed her to explore a different pathological context during post-doc, namely the tumour angiogenesis. Dr Camilli found jumping on this new field a difficult but exciting challenge!

In the third interview as part of the Early Career Innovators series, acknowledging the amazing translational work being done by early career researchers within the UCL Therapeutic Innovation Networks, Dr Gathoni Kamuyu highlights her Biologics TIN Pilot Data Fund awarded project “Identifying monoclonal antibodies for the treatment of Acinetobacter baumannii infections” and presents some advice for future applicants.

Please provide an overview of your Biologics TIN Pilot Data Fund awarded project. 

My project, “Identifying monoclonal antibodies for the treatment of Acinetobacter baumannii infections” will use single B-cell sequencing and cloning techniques to identify monoclonal antibodies targeting A. baumannii [3]. This will involve immunising mice to generate a robust antibody response against selected proteins, obtaining antigen-specific single B cells by fluorescence-activated cell sorting and screening each individual B cell for the production of antibodies effective in controlling the bacterial infection. Once a positive antibody-secreting B cell is identified, the corresponding monoclonal antibody it secretes, can be made in large quantities by recombinant protein expression [4, 5].

What is the motivation behind your project/therapeutic?

Acinetobacter baumannii has been referred to as the perfect predator in the media [6] and is number one on a recent WHO list of antimicrobial resistant (AMR) pathogens to which alternative therapies are urgently required [7, 8].

Through many different mechanisms, A. baumannii can survive and spread rapidly within hospitals, causes approximately 6-24% of nosocomial bacteraemia and pneumonia (particularly within intensive care units) and is associated with high morbidity and mortality rates [8-10]. Current treatment options uses complex antibiotic combinations to overcome the AMR profile, and there is an increase in reports on the incidence of infections caused by pan-drug resistant A. baumannii (non‐susceptibility to all agents in all antimicrobial categories) [11].

Monoclonal antibodies (MAbs) are a viable alternative to antibiotics that avoids the problem of drug resistance [12].  A carefully selected MAb offers multiple advantages over antibiotics that include rapid development with low toxicity, have minimal effect on the human microbiome, do not drive resistance to antimicrobials and can be conjugated to additional molecules to enhance antimicrobial effects.

Why did you want to apply to the Biologics TIN Pilot Data Fund?

The biologics TIN pilot data fund was specifically interested in funding projects by early career researchers (ECR), on biologics (including monoclonal antibodies), that were between the discovery and translational phases. This would allow the ECR to generate pilot data that could be used to apply for larger grants. My current research work had identified potential protein targets that elicited protective antibody responses against Acinetobacter baumannii making them ideal targets for monoclonal antibody development.

In addition, it was an opportunity to identify, interact and establish collaborations through the TINs, with groups within UCL that have similar research questions or have specialised research techniques.

What do you hope to achieve in the 6 months duration of your project?

I hope to have identified a subset of monoclonal antibodies targeting A. baumannii for further validation. I would also like to establish the pipeline I would use to identify monoclonal antibodies against additional antigens of interest that we would identify.

What are your next steps from now?

Hit the lab and generate data…

Do you have any top-tips for applicants currently going through the application process for the other TIN Pilot Data Funds?

Read the application instructions carefully, keep within the word count on your application and keep your 2 min pitch, simple, straightforward and to the point.

Learn more about and join the TINs

About Dr Gathoni Kamuyu

Dr Gathoni Kamuyu obtained a BSc. in Biochemistry (1^st Class Honours, University of Nairobi, Kenya) and MSc. in Molecular Biology of Infectious Diseases (Distinction, LSHTM,United Kingdom). Dr Kamuyu’s research career started at the KEMRI-Wellcome Trust Research Programme (Kilifi, Kenya), evaluating the link between exposure to parasitic central nervous system infections and epilepsy [1].

In 2017, Dr Kamuyu obtained a PhD from the Open University/KEMRI-Wellcome Trust programme, which focused on identifying the targets of protective antibodies against Plasmodium falciparum, one of the causative agents for Malaria [2]. During her initial post-doctoral training at University Hospital Heidelberg, Germany, Dr Kamuyu used in vivo models to evaluate a panel of P. falciparum proteins as targets of protective antibodies.

Currently, Dr Kamuyu is a Research Fellow within Prof. Jeremy Brown’s group in the Department of Respiratory Medicine, Centre for Inflammation and Tissue Repair (CITR), UCL. Her research focus includes understanding acquired immunity to Acinetobacter baumannii (A. baumannii), identifying the potential targets of protective antibodies and the mechanisms employed by A. baumannii to evade the effector functions mediated by the complement system.

References

1. Kamuyu, G., et al., Exposure to multiple parasites is associated with the prevalence of active convulsive epilepsy in sub-Saharan Africa. PLoS Negl Trop Dis, 2014. 8(5): p. e2908.

2. Kamuyu, G., et al., KILchip v1.0: A Novel Plasmodium falciparum Merozoite Protein Microarray to Facilitate Malaria Vaccine Candidate Prioritization. Front Immunol, 2018. 9: p. 2866.

3. Lu, R.M., et al., Development of therapeutic antibodies for the treatment of diseases. J Biomed Sci, 2020. 27(1): p. 1.

4. Carbonetti, S., et al., A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning. J Immunol Methods, 2017. 448: p. 66-73.

5. von Boehmer, L., et al., Sequencing and cloning of antigen-specific antibodies from mouse memory B cells. Nat Protoc, 2016. 11(10): p. 1908-1923.

6. Patterson, S.S.a.T., The Perfect Predator: A Scientists’s Race to Save Her Husband from a Deadly Superbug: A Memoir.

7. (WHO), W.H.O., Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotic. 2017.

8. Tacconelli, E., et al., Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis, 2018. 18(3): p. 318-327.

9. Allegranzi, B., et al., Burden of endemic health-care-associated infection in developing countries: systematic review and meta-analysis. Lancet, 2011. 377(9761): p. 228-41.

10. Cerceo, E., et al., Multidrug-Resistant Gram-Negative Bacterial Infections in the Hospital Setting: Overview, Implications for Clinical Practice, and Emerging Treatment Options. Microb Drug Resist, 2016. 22(5): p. 412-31.

11. Magiorakos, A.P., et al., Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect, 2012. 18(3): p. 268-81.

12. McConnell, M.J., Where are we with monoclonal antibodies for multidrug-resistant infections? Drug Discov Today, 2019. 24(5): p. 1132-1138.

In the second interview as part of the new Early Career Innovators series, acknowledging the amazing translational work being done by early career researchers within the UCL Therapeutic Innovation Networks, Dr Anais Cassaignau highlights her Biologics TIN Pilot Data Fund awarded project “Developing an scFv binder against nascent huntingtin” and presents some advice for future applicants.

Please provide an overview of your Biologics project.

This project entitled “Developing an scFv binder against nascent huntingtin” is looking to exploit the unique features of nascent proteins, i.e. the shapes they form while they are being made. I am currently pursuing the novel disease angle that is the focus of this award.

Relative to the fully formed protein, the nascent protein is typically protected against misfolding /aggregation. We are looking to show that this entity may be a tractable therapeutic target in Huntington’s Disease.

What is the motivation behind your project/therapeutic?

I am interested in understanding how proteins fold while they are being synthesised by the ribosome, and how the ribosome itself regulates and modulates this process¹. The correct folding of proteins in the cell is vital to all forms of life, and scientists are increasingly recognising that many diseases bear protein misfolding hallmarks including devastating neurodegenerative illnesses, several cancers and also diabetes.

Huntington’s is a devastating neurodegenerative disease, designated as an incurable disease with only symptomatic treatment currently available, and which often involves invasive delivery e.g. via spinal chord injections.  This is despite seminal work in the field that underpins much of what we understand regarding the pathological underlying processes and in particular how the causative agent, huntingtin, forms aggregates. I hope to be part of devising new therapeutic strategies that involve targeting the mutant form of huntingtin at the earliest point of biosynthesis – an angle which has not previously been explored in this manner.

Why did you want to apply to the Biologics TIN Pilot Data Fund?

I wanted to initiate a crucially needed orthogonal extension to the research I have been undertaking; building upon the wealth of collective knowledge that the entire lab and myself have been building together over years about how proteins are made and how they fold, and applying these paradigms to develop relevant disease-related models.

What do you hope to achieve in the 6 months duration of your project?

I want to demonstrate that targeting a nascent protein is possible, through binding an antibody and scFv to a nascent huntingtin during biosynthesis and monitoring how this modulates the folding/misfolding outcomes for this protein.

What are your next steps from now?

Finessing of assays and the production of samples of the nascent huntingtin. The protein will be translationally-arrested (a “snapshot” of biosynthesis) and then we will test the interaction of our antibody and scFv to it, and see how this influences the fate of this aggregation-prone protein.

Do you have any top-tips for applicants currently going through the application process for the other TIN Pilot Data Funds?

I would strongly encourage prospective applicants to reach out to the members of their respective TIN as the first step; their expertise will help you to appropriately refine your initial ideas and define the key questions in order to apply. Finally… Make a list of all the things you don’t know and read about them one by one.

Join the UCL Therapeutic Innovation Networks

About Dr Anais Cassaignau

Dr Cassaignau became interested in protein folding on the ribosome during her final year of BSc Biochemistry at UCL. Following this, Dr Cassaignau initiated a project within the Research department of Structural and Molecular Biology and has not left since, undertaking a Wellcome Trust-funded PhD and postdoc with John Christodoulou.

1. How does the ribosome fold the proteome? Cassaignau, AME, et al Ann. Rev. Biochem, 2020, 89, 389-415.  https://www.annualreviews.org/doi/abs/10.1146/annurev-biochem-062917-012226 

In the first interview as part of the new Early Career Innovators series, acknowledging the amazing translational work being done by early career researchers within the UCL Therapeutic Innovation Networks, Dr Giulia De Rossi highlights her Biologics TIN Pilot Data Fund awarded project “LRG1 antibody for diabetic macular oedema” and presents some advice for future applicants.

Please provide an overview of your Biologics project. 

There are currently 4.8 million people in the UK living with diabetes and over 300,000 of these have their sight threatened by a severe ocular complication called diabetic macular oedema (DMO), which has now become the most common cause of blindness in the working population.

Clinical studies have shown that leucine-rich alpha-2-glycoprotein 1 (LRG1) is enriched in the eyes of diabetic patients and we showed in other settings that it can drive vascular dysfunction. My hypothesis is that LRG1 is an early pathological switch in DMO and I believe that it may represent a novel/alternative pathway we could target therapeutically.

My project “LRG1 antibody for diabetic macular oedema” will test the efficacy of function-blocking antibody against LRG1 using murine models of diabetes. Specifically, I will be looking at the effects of this biologic on vascular homeostasis and permeability.

 

What is the motivation behind your project?

Currently, if you are diagnosed with DMO, you will receive monthly intra-ocular injections of VEGF-neutralizing antibodies. Unfortunately, this line of treatment has only a 50% chance of working and often responses are short-lived. As a result, despite the NHS spending £116m/annum, 2000 people go blind every year.

With diabetes reaching epidemic proportions, there is an urgent unmet need and market for developing new treatments for this devastating condition.

I believe targeting LRG1 with a function-blocking antibody has the potential not only to treat patients who are refractory to current therapies, but also to achieve earlier and therefore better outcomes in all patients.

Why did you want to apply to the Biologics TIN Pilot Data Fund?

I had an interesting set of preliminary data that I felt was suitable for applying for a pilot proof-of-concept grant.

The TIN pilot data fund was also my first opportunity to apply for and manage a grant as the lead applicant, which I hope will be a first stepping stone towards independent research and career development.

What do you hope to achieve in the 6 months duration of your project?

With the funding requested I plan to answer 3 critical questions: 1) Is LRG1 a pathological switch in DMO? 2) Is a Lrg1-deficient mouse protected from DMO? and 3) Can anti-LRG1 antibodies prevent the onset of DMO?

What are your next steps from now?

My planned experiments will support the preclinical dataset necessary to take the anti-LRG1 antibody into clinical trials, by establishing whether LRG1 is a valid target, and, together with the available supporting literature from clinical studies, will constitute the foundation for a translational grant application next year.

Do you have any top-tips for applicants currently going through the application process for the other TIN Pilot Data Funds?

Make sure you clearly describe: what the problem is, your proposed solution, why your approach is better, what you want to do next if successful (clear career and translational path).

As with every application with a limited word count, you might end up with an extremely abridged version of your initial text and some key concepts might become cryptic. Have someone from a different field review your application and tell you if they can still understand the key message you want to convey.

There is no space to describe the experiments in detail, so just explain the scientific questions you want to answer, you will have the opportunity to wear your scientist hat if you get to the Q&A stage.

Good luck future applicants!

Dr Giulia De Rossi will be discussing her Biologics TIN Pilot Data Scheme application process experience in the upcoming ACCELERATE Success event, “Grant Writing for Translational Research” on Tuesday 6th October.  This is an educational, translational training event to help UCL researchers write impactful applications by recognising the important elements of a translational research/innovation grant application and increase chances of funding success. Register here.

About Dr Giulia De Rossi

Dr Giulia De Rossi is a research fellow at the UCL Institute of Ophthalmology in the lab led by Professor Stephen Moss and Professor John Greenwood. Dr De Rossi’s training was in Biotechnology and the main focus of her PhD and post-doctoral work has been understanding the mechanisms underpinning vascular dysfunction and new blood vessel formation.

Dr De Rossi joined UCL in July 2019 to work on a Diabetes UK-funded project aimed at identifying new targets and mechanisms to treat the microvascular complications of diabetes.