Gene therapies rely on engineered virus carriers as vehicles for the delivery of synthetic genes that allow correction of disease-altered changes in multiple organs of the human body. Viruses exploited in gene therapy approaches have been modified to remove harmful properties and carry the therapeutic gene of interest. Multiple gene therapy programmes are currently undertaken in research laboratories using relatively small-scale production of viruses which enables optimisation of the doses and administration routes as well as testing for the safety and therapeutic efficacy of interventions in animal models of disease. However, facilities required for the production of large quantities of clinical-grade viruses of consistent quality-controlled GMP grade are rare in the UK. Thus, it can easily take several years before clinical trials can be conducted. Currently, existing facilities cannot meet the escalating demand of academically-led research needs for clinical-grade virus carriers. This is significantly obstructing numerous UK-funded world-leading disease-modifying discoveries to be translated into clinical trials for human benefit. The lack of suitable GMP facilities seriously hinders the development of much-needed novel effective treatments for multiple incurable diseases which cannot be treated by conventional drug compounds. We propose to address the manufacturing shortage by creating a Gene Therapy Innovation and Manufacturing Centre (GTIMC) which includes provision of a new state-of-the-art GMP manufacturing facility to support gene therapy projects emerging from UK universities. GTIMC will also support the development of improved viral vectors, improved yield from the manufacturing process and will also provide essential regulatory and training support. Moreover, the GTIMC hub will allow new training and high-skilled employment opportunities through the ShefVec facility itself, and future start-up companies.

The project seeks to exploit molecular biology to construct designer viruses that are structurally adapted for simple affinity purification and hence easier manufacture. The Gene Medicine web site shows that viruses are currently used in two-thirds of all gene therapy trials, with retro- and lentiviruses constituting -40% of these. Chromatography is preferred for vector purification and ion-exchange has been used in the multi-step purification of a lentiviral vector for a Phase 1 clinical trial. Precedence in protein manufacturing shows that such low specificity processes are now almost entirely superseded by affinity processes and this preference will likely emerge in virus manufacturing in order to exploit the resolution and simplicity of affinity chromatography. Recognising this we engineered a novel packaging cell line, Bio-293T, that metabolically produces an affinity-tagged lentivirus and we have demonstrated the enormous efficiency savings and flexibility offered by affinity capture for virus concentration. This first generation Bio-Lentivir packaging cell line has highlighted the inadequacy of conventional matrices for virus affinity purification, the critical need to optimise affinity-tag density on the viral envelope for efficient recovery and the potential of novel macroporous adsorbents to provide simple, single-step processing. It has also shown that biotin-tagged lentivirus can be complexed with streptavidin paramagnetic particles, resulting in the most efficient isolation and concentration method yet described for lentiviral vectors. However, most clinical applications require free virus, for which use the paramagnetic nanoparticle capture approach is unsuitable without virus elution, which, for this high affinity system leads to low process yields. Following our proof-of-concept there is consequently a need for a second generation Bio-Lentivir packaging cell line designed to overcome these limitations and fully exploit the potential for the efficient production of high titre, highly purified, clinical grade retroviral and lentiviral gene transfer vectors.The project outputs will be novel His-tag/Bio-Lentivir packaging cell lines, new adsorbent materials for virus purification and an integrated virus production scheme. These objectives stem from the joint experience of the Kings and Cambridge teams on the successful development of the Bio-293T packaging cell line and will be addressed by bringing together expertise in molecular biology, vector design and packaging cell line construction (at Kings) with bio-materials, virus and affinity processing expertise (at Cambridge).

The UK government's support for the Life Sciences Industry Strategy (Bell Report, 2017) recognises the importance of developing new medicines to facilitate UK economic growth. Examples include new antibody therapies for the treatment of cancer, new vaccines to control the spread of infectious diseases and the emergence of cell and gene therapies to cure previously untreatable conditions such as blindness and dementia. Bioprocessing skills underpin the safe, cost-effective and environmentally friendly manufacture of this next generation of complex biological products. They facilitate the rapid translation of life science discoveries into the new medicines that will benefit the patients that need them. Recent reports, however, highlight specific skills shortages that constrain the UK's capacity to capitalise on opportunities for wealth and job creation in these areas. They emphasise the need for 'more individuals trained in advanced manufacturing' and for individuals with bioprocessing skills who can address the 'challenges with scaling-up production using biological materials'. The UCL EPSRC CDT in Bioprocess Engineering Leadership has a successful track record of equipping graduate scientists and engineers with the bioprocessing skills needed by industry. It will deliver a 'whole bioprocess' training theme based around the core fermentation and downstream processing skills underpinning medicines manufacture. The programme is designed to accelerate graduates into doctoral research and to build a multidisciplinary research cohort; this will be enhanced through a partnership with the Synthesis and Solid State Pharmaceutical Centre (SSPC) and the National Institute for Bioprocess Research and Training (NIBRT) in Ireland. Research projects will be carried out in partnership with leading UK and international companies. The continued need for the CDT is evidenced by the fact that 96% of previous graduates have progressed to relevant bioindustry careers and many are now in senior leadership positions. The next generation of molecular or cellular medicines will be increasingly complex and hence difficult to characterise. This means they will be considerably more difficult to manufacture at large scale making it harder to ensure they are not only safe but also cost-effective. This proposal will enable the CDT to train future bioindustry leaders who possess the theoretical knowledge and practical and commercial skills necessary to manufacture this next generation of complex biological medicines. This will be achieved by aligning each researcher with internationally leading research teams and developing individual training and career development programmes. In this way the CDT will contribute to the future success of the UK's bioprocess-using industries.

We have developed a novel gene expression technology that has applications in the R&D kits market and in the manufacture of biopharmaceuticals. We plan through this current study to benchmark our expression technology against those systems currently available in the marketplace. This study will showcase the key areas of technical superiority of our technology, inform its subsequent positioning in the market place, inform our business planning and act as an enabling primer for private equity investment. The outputs from this grant will lead to partnering collaborations with industry, provide impact for BBSRC funded research, and potentially lead to wealth generation and job creation for the UK.

The Kings/Royal Free/UCL Gene Therapy Innovation Hub will manufacture clinical-grade gene therapies for the UK academic and clinical community. This will allow promising treatments for a wide range of rare and common diseases to be tested in patients. The results from these early clinical studies may then support larger-scale trials and ultimately new therapies for patients. Provision of suitable quality (GMP) gene therapy product is a key limiting factor for progress in this exciting field. Our Hub will address this directly through a major increase in UK capacity. This will cover the major types of gene therapy used, adeno-associated virus (AAV) and lentivirus, as well as gamma-retrovirus. In addition, we will invest in developing new approaches to increase the amount of gene therapy product that can be made at one time (in one batch). This will mean that each trial needs fewer batches, and that applications needing high doses (for example, administration systemically) become possible. It will also reduce costs of manufacture. We will also work to increase the UK's overall capabilities to manufacture gene therapies, through creation and provision of dedicated training courses, both online and in person, at a variety of levels, to suits varying needs of different groups. In addition to this, we will lend our expertise to other developing Gene Therapy Innovation Hubs, to help them to become operational as quickly as possible. King's, Royal Free and UCL offer unmatched expertise in the UK in gene therapy manufacturing, and we are ideally-placed to contribute to the UK's success in improving therapies for many poorly-treated diseases, and generating sustainable economic benefit for the country.