Impact case study database

1) Stimulation of innovation and entrepreneurial activity by industry

Following a competitive process, a collaborative project on ‘Therapeutic activators of TREK-2 K2P channels’ was adopted by LifeArc’s Centre for Therapeutic Discovery in March 2019 [a]. LifeArc stated: ‘University of Kent and LifeArc have entered into a collaboration agreement on 6 March 2019 with respect to activators of TREK-2 potassium channels for the treatment of chronic pain under the direction of Prof Mathie and Dr Paul Wright’ [b]. Furthermore, LifeArc ‘will conduct target validation, high throughput screening and will further develop any “hits” identified through that screening’ [a]. This comprised an initial commitment from LifeArc of around £600k internal expenditure and £200k external expenditure [b, c]. In accordance with the agreed plan of work, LifeArc had, by October 2020, ‘carried out thallium flux screens of the LifeArc Diversity Set (100K compounds) and identified a number of hit compounds for further analysis’ [c]. They ‘studied the detailed chemical and biological properties of these compounds and commissioned pharmacokinetic profiling of them’. ‘The “in vivo” efficacy of a representative lead compound in models of pain has been confirmed through investment in studies by an independent external investigator.’ Through this collaboration, LifeArc ‘have already identified at least one family of novel TREK-2 channel activators as potential therapeutic agents to treat pain’ [d].

This project has also directly led to the ‘refinement of existing assays used within LifeArc to optimise identification of K2P channel activator compounds’ [d]. These developments have been published for the benefit of other research organisations working in this area [R2, R6, d]. In September 2020, LifeArc (with the University of Kent) were awarded an Industrial Fellowship from the 1851 Royal Commission of the Exhibition linked to this project [e]. The award covers salary costs for the fellow and other expenses and ‘plays a crucial role in facilitating the relationship between institutions and industry in the UK, offering highly valued funding for research and development’.

2) New products recommended, repurposed, and adopted for use

Two compounds identified through the University of Kent team’s collaborative research with LifeArc (terbinafine, since 2017; and pranlukast, since 2019) have been repurposed for use and are now sold by Biotechne and associated companies (2,200+ employees worldwide) as TASK-3 and TREK-2 channel activators, respectively. For both compounds, Biotechne cite the Kent team’s original research with LifeArc [R2, R6] in their catalogue and product datasheets [f], as sole primary evidence for K2P channel activation by these compounds.

A related collaboration with Pfizer and Icagen identified GI-53059 [R5], which has been recommended by the International Union of Basic and Clinical Pharmacology’s (IUPHAR) authoritative Guide to Pharmacology as a selective activator of TREK K2P channels. Since 2018, GI-53059 has been sold commercially as a TREK channel activator by several international pharmaceutical companies (Biotechne, Glixx Biologicals, Probechem, and MedKoo). In catalogues and product datasheets [g], the University of Kent team’s original research results [R5] are cited as sole, primary evidence for the mechanism of action of GI-53059. Since 2019, GI-53059 and pranlukast have been included in Biotechne’s Tocriscreen 2.0 compound library as ‘best in class’ biologically active chemical tools [f, g].

3) Transformation in patient treatment

The Kent team’s research has led to a new treatment for patients suffering from BBMRDS. Fenamate compounds, which the team identified as activators of the mutated TASK-3 K2P channels seen in BBMRDS [R1], have been repurposed, as a direct result of the research, for use in this condition. This is the only treatment used for BBMRDS to date. In a 2016 publication, Dr John Graham and colleagues stated: ‘Treatment of one Brazilian child with FFA ointment has been attempted without any obvious complications. Treatment of other patients with the drug MFA (Rx Ponstel) is currently underway in the USA with no apparent adverse effects. Patient 1 has been on MFA for one year, and his development and responsiveness is clearly better while on MFA than while off it during treatment rest periods. Patient 3 has been on MFA for 6 months, and he experienced improved developmental milestones about 4 months after starting the drug’ [h].

Since taking MFA, a patient in the UK has ‘made remarkable progress and now reached the ceiling of the Hammersmith motor scale’ [i]. The Director of Personalised Care and Professor of Clinical Pathology and Pediatrics at the Children’s Hospital, Los Angeles, states: ‘Four years of data is available for one patient. MFA was started at 15 months of age. Like other treated patients, he had an increase in energy and stamina while on MFA. At 24 months of treatment, he had significant improvement [in tone and motor outcomes] and has continued to improve by 5.5y.’ The Director also confirms that ‘without the pioneering experiments of Dr Veale and Prof Mathie, we would never have had the insight to test this promising treatment for Birk-Barel patients. Both clinicians and families are in their debt.’ [j].

Transforming research strategies and priorities at the European Commission Joint Research Centre, Diamond Light Source Ltd, and the University of Oldenburg

The European Commission’s Joint Research Centre, Diamond Light Source Ltd, and the University of Oldenburg have altered their research strategies and priorities as a result of collaborative work carried out with Sumbayev and colleagues on the AML project [a, b, d].

1. European Commission’s Joint Research Centre

The Joint Research Centre (JRC) is the European Commission's science and knowledge service, which employs scientists to carry out research in order to provide independent scientific advice and support to EU policy. Between 2017 and 2019, the JRC collaborated with Sumbayev on the AML project. Dr Luigi Calzolai, a scientist working on nanobiotechnology at the JRC (Italy), affirms that ‘This work had a strong impact on the research priorities of our institution and we invested funding and resources into this work’ [a]. Calzolai goes on to explain that the JRC designed and provided gold nanoparticles that Sumbayev and his group used to develop functional nanomaterials to specifically recognise human acute myeloid leukaemia cells [a]. The collaboration with Sumbayev resulted in several projects on which the JRC is now working. Calzolai confirms that ‘These projects are dealing with development of functionalised nanomaterials in order to recognise specific cells and deliver specific compounds into them (starting in 2017 and running until now [November 2020]). This includes also development of novel and advanced techniques for characterisation and design/optimisation of biocompatible functional nanomaterials. This work included financial investments [c. 140,000 euros] and involvement of human resources – two postdoctoral scientists’ [a].

1. Diamond Light Source Ltd

Sumbayev and colleagues have collaborated with Diamond Light Source Ltd [b, c], which runs the UK’s national synchrotron facility. Diamond’s circular dichroism Beamline (B23), which enables structural, functional, and dynamic interactions in materials such as proteins, nucleic acids, and chiral molecules to be observed, was used in the AML project to investigate interactions between key proteins involved in the immune evasion pathway. Dr Rohanah Hussain (Senior Beamline Scientist at Diamond) states that ‘This work [carried out since 2016] had a significant impact on our research strategy and appeared to be one of the major research highlights of Diamond Light Source during the last two years’ [b].

In 2019, Beamline B23 initiated a new project which includes the development of functional non-toxic peptides to target protein kinase C in order to block exocytosis of immunosuppressive proteins by AML and other cancer cells. Research carried out by Sumbayev led Diamond Light to start the project and revise their research plans [b]. Hussain confirms that ‘As a result of knowledge obtained via Dr Sumbayev’s research, we started a fundamentally novel project here at Diamond Light Source Ltd. We have been developing peptides for the purpose of blocking the enzyme called protein kinase C (PKC) which facilitates exocytosis. Dr Sumbayev’s research demonstrated that PKC is involved in the latrophilin-1 triggered immune evasion pathway operated by human cancer cells’ [b]. Diamond has already invested around £60,000 into the new project to cover the costs of peptide design and characterisation of their ability to bind and inhibit PKC. Hussain explains how the long-term idea is to design cell-penetrating peptides to block the immune evasion pathway, allowing cytotoxic lymphoid cells to display maximal anti-cancer activity [b].

1. The University of Oldenburg

Dr Bernhard Gibbs, Principal Research Associate and Reader in Experimental Allergology at the University of Oldenburg in Germany, has collaborated with Sumbayev and colleagues on research into acute myeloid leukaemia since 2013. Dr Gibbs provided primary human blood samples obtained from healthy donors. He also shared his expertise on the use of primary human blood samples and helped with the ethics documentation. Dr Gibbs confirms that ‘the research work of Dr Vadim Sumbayev, to which our institution has contributed, had a significant impact on our research strategy’ [d]. As a result of the discovery of the latrophilin-1-induced immunosuppressive pathway operated by AML cells [R1-R6], Gibbs confirmed in 2020 that the Division of Experimental Allergology and Immunodermatology have revised their research strategy to include the role of immunosuppressive pathways in cancer and autoimmune diseases. Furthermore, Gibbs confirmed that ‘the role of this pathway in AML and other cancers is also being considered as a new research topic in one of our Faculty research priority areas (“Potentialbereich Onkologie”), which is currently being established’ [d].

Facilitating changes in medical screening procedures

The findings of the Kent research team and collaborators have had a significant impact on the medical screening procedures at two hospitals. The University Medical Centre Hamburg-Eppendorf (Germany) and the University Hospital of Bern (Inselspital), Switzerland, have changed their sample screening practice for patients with leukaemia and paediatric tumours [e, f]. Professor Walter Fiedler (MD) from the University Medical Centre Hamburg-Eppendorf states: ‘As a result of our collaborative research [with Sumbayev since 2017] our department amended patient sample screening procedures detecting latrophilin-1 and galectin-9 in them in order to characterise the disease’ [e]. In 2019, Inselspital Bern also amended their screening procedures of paediatric tumour samples in the hospital as a result of collaborative work led by Sumbayev. Dr Elizaveta Fasler-Kan from the Department of Paediatric Surgery explains that they have found that galectin-9 and its receptor Tim-3 are highly present in paediatric tumours, including, but not limited to, Wilms tumours. Fasler-Kan states: ‘Currently, we are screening paediatric tumour samples for expression/levels of Tim-3 and galectin-9 in order to estimate their ability to escape immune surveillance’ [f].

Informing teaching programmes for medical and biomedicine students

Two universities (Oldenberg and Basel) and a medical school (Inselspital, Bern) have changed their education programmes for students to include the findings of the Kent team’s research [d, f, g].

Professor Gibbs at the University of Oldenburg confirms that ‘the research work of Dr Vadim Sumbayev […] has had a significant impact on our […] teaching programmes’ [d]. Gibbs describes how the Division of Experimental Allergology and Immunodermatology has revised its teaching programmes on immunology and anti-cancer immunity to include the latrophilin-1-induced immunosuppressive pathway, giving the example of latrophilin-1 as a potential biomarker for AML diagnostics [d]. Gibbs states: ‘This includes undergraduate medical students doing electives in our Division as part of “Longitudinales Forschungscurriculum” (which took place in both summer and winter semesters since 2019), as well as a new international M.Sc. course, which has just started, in Molecular Biomedicine (currently involving 14 students)’ [d].

Sumbayev’s discovery is also having an impact on the teaching programmes for Biomedicine students at the University of Basel (Switzerland) [g]. Dr Fasler-Kan confirms that ‘Each year group includes 100 students and since 2018 we have included the Tim-3-galectin-9 immune evasion pathway into the Basic Immunology course when covering aspects of anti-tumour immunity and immune evasion pathways’ [g]. Inclusion of Sumbayev’s findings is helping students to better understand the severity of blood cancers and the inability of the immune system to deal with them [g]. Fasler-Kan explains how teaching about the pathway helps to illustrate the power of anti-cancer immune evasion machinery to impair the activity of immune competent lymphocytes leading to disease progression [g]. Publications from the Kent team and a sound recording of a webinar given by Sumbayev at the 23^rd World Congress on Advances in Oncology in 2018 are used as teaching materials on the course [h].

Dr Fasler-Kan also states that the research work of Sumbayev and colleagues has influenced teaching programmes and led to the inclusion of Kent’s discovery in teaching programmes for medical students at Inselspital Bern [f]. Fasler-Kan confirms that ‘we have included [Sumbayev’s] findings in our Advanced Immunology programme for medical students (undergraduate) – roughly 100 students’ [f]. Kent’s research is helping students at Inselspital to understand the concepts of tumour immune evasion and tumour associated antigens. Fasler-Kan also confirms that students are being taught about latrophilin-1 as an example of a biomarker that is expressed in the cancer cells of the vast majority of AML patients [f].

1) Rapid development of reference standards for Ebola, Lassa Fever, MERS-CoV and SARS-CoV-2 using pseudotype technology

The National Institute for Biological Standards and Control (NIBSC) has identified pseudotypes as a technology that can facilitate the development of accredited reference standards in a significantly shorter timeframe [a], which is particularly important for emerging viruses. Collaboration between Temperton and NIBSC, initiated in 2015, resulted in the development of WHO-accredited nucleic acid and antibody reference reagents for Ebola [b]. Mark Page (Head of the Emerging Viruses Group at NIBSC) states that ‘most of the WHO R&D Blueprint are pathogens in hazard group 3 or 4’, and that Temperton’s PV have been identified by NIBSC ‘an essential tool for the characterisation of candidate reference reagents, alleviating the need to work at a high containment level, which for Ebola would not have been possible as NIBSC does not possess a CL4 laboratory’ [a]. Temperton’s PV-based neutralisation assays were critical for the evaluation of the Ebola plasma standard and PV were essential for production of the nucleic acid test (NAT) standards that are made by packaging the Ebola genome inside Temperton-designed retroviral pseudotype particles. Without access to Temperton’s pseudotype technology, these standards would have taken significantly longer to develop and release. Page confirms that ‘The use of pseudotype viruses for the development of reference material has now been adopted by default at NIBSC and permits the production of standards to a responsive and fluent timeframe for the priority pathogens listed in the WHO R&D Blueprint’, and further states that ‘The impact of this is that NIBSC has been made an implementing partner with Coalition for Epidemic Preparedness Innovations (CEPI) […] to produce antibody standards to enable vaccine development against priority pathogens’ [a]. In February 2019, NIBSC, enabled by Temperton’s pseudotype technology, leveraged contracts from CEPI for 1m USD to produce standards for Lassa fever virus [a], and, in collaboration with Temperton, NIBSC have also produced reference reagents for MERS-CoV and SARS-CoV-2 ‘with an accelerated time frame’ [R6] [a]. Page concludes that ‘There will always be the potential for […] outbreaks that could have global impact and the VPU and NIBSC alliance is ideally suited to respond to the next emergency’ [a].

2) Employing SARS-CoV-2 pseudotypes as direct assay standards for clinical diagnostic tests

In 2020, UKRI funded the Humoral Immune Correlates of COVID-19 (HICC) consortium, which is comprised of a core of founder members in receipt of direct funding (University of Cambridge, Kent Viral Pseudotype Unit. MAP Diagnostics, Royal Papwoth and Addenbrookes NHS Trusts) and a large network of collaborators [G3], to carry out SARS-CoV-2 research into immune correlates. In 2020, a collaboration between Temperton and the diagnostics company MAP Diagnostics (Bedford, UK), which develops novel diagnostic tests based around MALDI-ToF mass spectrometry, was initiated via HICC [c]. This collaboration involves the use of Temperton’s SARS-CoV-2 pseudotypes directly as assay standards. MAP Diagnostics is developing SARS-CoV-2 clinical diagnostic tests based on direct MALDI-ToF mass spectral analysis of gargle/saliva samples, and Temperton’s pseudotypes have been incorporated into the company’s protocols, enabling safer and rapid methods development at reduced cost [c]. Professor Ray Iles (CEO of MAP Diagnostics) states: ‘The COVID-19 pandemic prompted the development of a mass spectral technique for identification of viral envelope proteins direct from gargle/saliva samples. This would not have been possible without the use of SARS-2 pseudo-type, and other corona viral pseudo-types, developed by Dr Temperton’s research’ [c]. The timely availability of ‘safe to manipulate mock [pseudotype] virus; with the same biochemical, conformational and physical biology of the target proteins in situ within a viral-like envelope exosome, allowed not only the spectral but pre-processing biochemistry to be developed in a non CAT3 restrictive laboratory setting’ [c]. Isles further states that their protocol is now ‘being adopted by other mass spectral analysis centres’, and the company is sending Temperton’s pseudotypes ‘to global collaborators as a quality and positive control to be run with every diagnostic test’ [c]. In addition to using Temperton’s SARS-CoV-2 pseudotypes, Isles states that, via collaboration with Temperton, ‘Other viral pseudo-types are being tested in order to discover the […] “spectral fingerprint” of virus infections’, and highlights that ‘the cost, speed and utility of the technique is going to have major impacts on global health care and disease diagnosis beyond the COVID-19 pandemic’ [c]. The assays used by HICC and MAP Diagnostics are all calibrated using the NIBSC SARS-CoV-2 reference reagents [R6].

3) Applying the pseudotype virus platform to vaccine research and development for influenza, SARS-CoV-2, Marburg and Lassa Fever

Temperton has a longstanding collaboration with VisMederi (Siena, Italy), which undertakes serological assays for pharma and towards licensure of influenza, coronavirus, and other vaccines [d]. Temperton’s influenza and SARS-CoV-2 neutralisation assays and know-how have been successfully translated to VisMederi via this collaboration, and in 2016 a new PV-based enzyme-linked lectin assay (PV-ELLA) was co-developed that enables the measurement of responses to neuraminidase (NA) in influenza vaccines [d]. This new assay is highlighted by CEO Professor Emanuele Montomoli: ‘In 2016, a VisMederi researcher, Fabrizio Biuso, joined Nigel Temperton’s laboratory for refining the […] pseudotypes platform. In particular the project […] aimed to study the development of [an] ELLA assay […] to evaluate human anti-NA antibodies’ [d]. According to Montomoli, the transfer of Temperton’s PV technology ‘allowed VisMederi to execute several clinical and preclinical studies and it was […] applied to Flu, Rabies and now [in 2020] it will be useful […] in SARS CoV-2 studies in order to accelerate the development of SARS-CoV-2 assays’ [d]. Montomoli states that working with PV will ‘eliminate the need [… for] wild-type virus, meaning […] this assay can be performed at biosafety level II (BSL2)’, and that ‘R&D of this nature would be laborious and expensive to perform with the native virus’, confirming that ‘the collaboration with Nigel Temperton […] saved costs to the company and increased the staff safety level’ [d].

Temperton has been collaborating with DIOSynVax, a Cambridge University spin-out company, since 2017. Professor Jonathan Heeney, CEO of DIOSynVax, explains that DIOSynVax is focused on vaccine R&D using a technology platform that encompasses a computational approach ‘for the rapid selection of vaccine antigens that are able to elicit cross protective immune responses to an array of different viruses’ [e]. Temperton acted as a scientific advisor from September 2016, and sought to develop a trivalent vaccine to protect against Ebola, Marburg and Lassa Fever. Heeney states that ‘The project included successful animal trials and the vaccine has now moved to Phase 1 clinical trials’ [e]. Temperton’s PV-based serological assays form a key component of the company’s new technology platform, and scientists at DIOSynVax are routinely trained in PV development, production, and deployment with Temperton’s support [e]. Heeney states that ‘The PV assays are integral to the vaccine development pipeline as the data is fed back into machine learning processes to improve vaccine antigen selection’, and that ‘R&D of this nature would usually be laborious and expensive to perform with the native virus, however, our process design enables adaptability to new virus strains and is […] safer for staff to work with’ [e]. According to Heeney, results obtained using Temperton’s PV assays have underpinned ‘successful grant applications’ that have enabled the company to ‘move into new areas of vaccine R&D’. Current collaborative projects include vaccine development for influenza [G1, G2] and coronavirus, with the DIOS-CoVax vaccine having recently received funding to enter a Phase 1 clinical trial [e]. Protective Ab responses and critical readouts for clinical trials of COVID-19 vaccines including DIOS-CoVax are being addressed in a joint-funded NIHR project [G3].

In 2018, Temperton’s collaboration with Oxford University on the discovery of a new ‘universal’ influenza vaccine epitope, which involved the use of the Kent team’s PV-based neutralisation assays [R4], resulted in a US-based spin-out company, Blue Water Vaccines. The company’s vaccine program aims to commercialise a vaccine that could protect against all influenza strains and provide life-long immunity. Dr Craig Thompson (CEO at Blue Water Vaccines) states that ‘our relationship with Dr Temperton has centred around the pseudotyped virus assay technology his lab has developed […]. This technology has become central to the discovery of novel antigen targets as part of our influenza vaccine program’ [f]. Thompson confirms that the use of Temperton’s PV-based neutralisation assay has enabled the assessment of ‘protective neutralising antibody responses […] under CL2 conditions’, and that ‘Without this assay we would need to work under more restrictive CL3 or SAPO4 conditions using live influenza virus, which would be impossible to undertake at the scale we require. Consequently, our use of the […] assay has sped up the development of our influenza vaccine’ [f]. Thompson also states that the provision of laboratory assistance, constructs, and techniques developed by the Kent team ‘has been vital in expanding the range of influenza glycoproteins analysed’ by the company [f].

4) Enabling isolation and characterisation of antibodies with potential immune prophylaxis and vaccine potency application through the use of pseudotype neutralisation assays

Since 2017, Temperton has built a strong relationship with NIBSC Biotherapeutics, which discovers and develops single domain antibodies (sdAbs). They have been using Temperton’s influenza pseudotypes in neutralisation assays for first-line screening to identify broadly neutralising sdAbs with potential therapeutic, diagnostic, and vaccine potency application [R1, R3]. Live pathogenic viruses could not have been used in such assays owing to the exorbitant costs and safety constraints associated with the running of their R&D screening pipeline under high containment [g]. Through this partnership, Dr Hufton has additionally leveraged funding from the Biomedical Advanced Research and Development Authority (BARDA) in the US to ‘develop an influenza vaccine potency assay based on broad neutralising and lineage specific single domain antibodies. This “universal” assay will ultimately reduce the need for the seasonal generation of strain specific reagents and so speed up the influenza vaccine development pipeline.’ [g].