Shortage of transplantable livers Liver transplantation is a life-saving treatment for patients with liver failure. Sadly, 15% of patients on the waiting list either die, or become too unwell for a transplant, due to an organ shortage. Despite this, 2 out of 5 potential deceased donor livers are declined for transplant. Many of these declines are because the donors possess characteristics that make their organs too susceptible to deterioration during preservation. Improved preservation will increase the number of transplantable livers. How are livers preserved? The traditional method is on ice. During this period, the organ metabolises without oxygen, generating harmful substances that are released when the liver is implanted into the patient. Machine perfusion at body-temperature has a number of benefits over ice storage. The machine provides the liver with nutrients and oxygen to enable normal metabolism. This allows the organ to recover during preservation, resulting in less injury on implantation and more transplantable livers. Current liver perfusion devices are licensed up to 24 hours. Prolonged perfusion up to 7 days seems possible in the laboratory, but remains at the early development stage. The benefits of prolonged perfusion in transplantation Prolonged perfusion will improve transplant outcomes and increase organ utilisation. The additional time available will enable improved recipient preparation, planned day-time surgery and wider organ-sharing. It will facilitate more detailed assessment of organ quality to minimise discarding of viable livers, and delivery of treatments to resuscitate injured organs. Use of prolonged perfusion in liver failure Beyond transplantation, prolonged perfusion holds promise as a treatment for acute and acute-on-chronic liver failure. These are severe forms of liver injury and many of these patients do not survive. However, the liver possesses the ability to regenerate, meaning that if these patients can be supported temporarily, many will recover. Whereas patients with kidney failure can be supported with dialysis, an equivalent for liver failure has proven elusive due to the complex roles of the liver. However, livers retain many of these functions during machine perfusion. If a liver can be successfully perfused for enough time to allow liver regeneration, a non-transplantable liver could be used to support patients with liver failure to recovery, or to survive long enough to receive a suitable organ. Aims of the research 1) Improve machine perfusion of the liver at body temperature to enable prolonged perfusion to 7 days. 2) Assess whether a liver support system, based on prolonged machine perfusion of a liver, is capable of replacing the function of a failing liver. 3) Determine the best method to connect a patient to the liver support system. How will these be achieved? I will test three interventions targeted at prolonging machine perfusion: 1) Incorporation of a specialist filter to remove harmful substances from the perfusion blood and correct the salt levels. 2) Addition of a solution to the perfusion blood to rejuvenate old red blood cells. 3) Provision of fish-oil based fats with the nutrition for the liver. To determine whether a machine-perfused-liver is capable of replacing a failing liver, I will subject the liver to challenges that mirror those that occur in liver failure. Finally, we will compare two connection methods: one in which blood is directly exchanged between the "patient" (in this case, a second machine-perfused liver) and the support system, and a second in which specialist filters prevent cells mixing between the two circulations. Future Plans Prolonged perfusion will be combined with work exploring organ assessment and gene therapies during preservation to accelerate their development. The liver support experiments will lead to a study in pigs to determine whether the system is effective in practice.

Liver disease is the only major cause of death still increasing year-on-year and the number of people dying from liver disease has doubled in the last 20 years. Liver transplantation is a successful treatment for end-stage liver disease but is limited by donor organ availability. As a result of this donor shortage, 20% of patients are removed from, or die whilst on the UK waiting list every 2 years. It is therefore of great importance to identify ways of making more organs available for transplantation. One way of achieving this is through the utilisation of more "marginal" or suboptimal livers, accepting the potential risk of poorer outcomes. Fatty livers derive poor outcomes when transplanted and as a result, a large number are discarded. It is believed that fatty livers are particularly susceptible to the damage associated with cooling the organ for storage prior to transplantation. Obesity levels in the UK and worldwide are increasing at an alarming rate. With this, comes an increasing incidence of fatty liver disease and it is estimated that 33% of the UK population suffer from non-alcoholic fatty liver disease. An increase of fatty livers in the donor pool is therefore inevitable and identifying methods of optimising and salvaging these livers for transplantation is essential. The key to successful transplantation of fatty livers may lie in organ preservation. Current practice involves cooling the organ with ice and a specialised preservation solution, termed static cold storage (SCS). New technology involves maintaining the liver on a machine that provides oxygen and nutrition to the liver at normal body temperature, known as normothermic machine perfusion (NMP). We believe that NMP will allow the safe transplantation of fatty livers by: 1) avoiding the deleterious effects of cooling; and, 2) enabling the delivery of de-fatting agents to the perfusion circuit to remove the fat from the liver before it is transplanted. At present the University of Oxford are leading a multi-centre randomised controlled trial comparing SCS and NMP as a preservation technique. Several fatty livers have been transplanted as part of the study and they provide a unique opportunity to compare outcomes from fatty livers transplanted via the two preservation techniques and compare them to normal, non-fatty livers. As part of the study protocol, liver biopsies are obtained prior to and at the end of the preservation period and following implantation of the liver in the recipient. I will analyse these biopsies and compare the fat content at the three time points in each group. I will also be able to assess the degree of damage associated with cooling after transplantation and compare this for fatty livers in the SCS and NMP groups. Post-transplant patient and organ outcomes will be compared in each group. The results obtained will provide a unique insight into the effects of NMP on fatty human livers, data which has not been reported to date. I also plan to obtain 20 livers which have not been transplanted and discarded due to a high fat content. Tissue samples will be obtained and de-fatting agents which have previously shown to reduce the fat content of fatty rat liver cells will be validated and refined for human cells in a laboratory setting. Once the agents have been optimised, I plan to apply them to whole livers through the NMP circuit. By examining biopsy and perfusate samples from the circuit for fat content at specific time points during perfusion I will be able to establish the effect on NMP +/- de-fatting agents on fatty livers. I aim to gain an understanding of the process by which de-fatting occurs in the liver by investigating key processes of fat breakdown in the liver. Through the above experiments I plan to demonstrate that a combination of NMP and de-fatting agents will allow the safe and reliable transplantation of fatty livers. This will result in more livers being transplanted and ultimately, lives saved.

The greatest challenge in oncological drug delivery is achieving successful penetration and distribution of the therapeutic agent throughout the tumour: billions of pounds have been spent to date in deploying biochemical approaches in an attempt to solve what is essentially an engineering problem, namely the transport of therapeutics from the blood stream to reach every cancer cell. OxCD3 will seek to transform both clinical and industry practice in drug delivery by demonstrating the value and feasibility of engineering approaches, involving a combination of stimulus-responsive nanocarriers and medical devices already in clinical use, for improved tumour uptake and therapeutic outcome. The Programme Grant will enable the creation of a sustainable, world-unique multi-disciplinary environment for combinational engineering of biology, chemistry and medical devices to improve drug delivery under a single roof. It is also expected to create a unique training environment for the next generation of young scientists working on combination therapies and biomedical nanotechnology, by providing direct exposure to regulatory and manufacturing issues encountered when translating laboratory research into production and clinical practice. A unique feature of the Centre is the capability to design both devices and drug delivery vehicles under a single roof. In the first 5 years, under EPSRC funding, up to 3 carefully selected "Device+Drug" exemplars will be manufactured to GMP, ready for Phase I clinical trials, to provide compelling evidence of feasibility to industrial partners and clinicians; in the next 5 years, a private-public partnership will be built to complete clinical trials of these exemplars using therapeutics of strategic significance to the pharmaceutical industry; beyond 10 years, full industrial sponsorship of the OXCD3 is anticipated, which will focus on addressing next-generation challenges in drug delivery (beyond cancer) in partnership with industry and clinicians. The transformative aim over 50 years is to position the UK as the world leader for multi-disciplinary drug delivery development, complementing its existing position as a drug discovery leader, from design to manufacture and clinical trials.

Synthetic Biology is a growing field of science that combines Biosciences, Chemistry, Physics, Information Technology and Engineering, and involves the redesigning end engineering of organisms for functional purposes, for example to produce valuable substances (e.g. medicines) or gain new functions (e.g. sensing and responding to something in the environment). Synthetic Biology aspires to tackle grand challenges surpassing what is possible through traditional technologies: it has wide-ranging applications in healthcare, environmental protection, energy, agriculture, computing, advanced chemicals and materials. Synthetic Biology has grown significantly in the UK over the past decade, thanks to a >£400M investment via the Synthetic Biology for Growth Programme. One of the key investments has been the SynBioCDT: the first UK CDT in Synthetic Biology funded in 2014 by the EPSRC and BBSRC and run by the Universities of Oxford, Bristol and Warwick. The SynBioCDT trained 79 excellent PhD students selected from >650 applicants, and attracted support from industrial, academic and public-facing partners. Our graduate students have gone on to work within the bioeconomy and have established disruptive start-ups. The term "Engineering Biology" has been recently adopted to highlight the essential transition of Synthetic Biology into a mature Engineering discipline. The recent UKRI National Engineering Biology Programme (NEBP) sets the UK ambition for the field and encompasses the capabilities that can support the exploitation of Engineering Biology for economic and public benefit. The Universities of Bristol and Oxford aim to establish a new CDT in Engineering Biology, the EngBioCDT, to train the academic and industrial Engineering Biology leaders of tomorrow, and to equip them with skills needed to contribute toward scalable, robust, and transformative engineering of biomimetic and biological systems. The EngBioCDT builds on our experience with the SynBioCDT and will address the NEBP requirement for a new generation of biological engineers able to translate cutting-edge science into real-world impact; it will support the EPSRC focus area 'Frontiers in Engineering and Technology'. The EngBioCDT will enable cohesive cohorts of students to gain expertise in the design, modelling and engineering of biological components and systems; to understand broad concepts ranging from biomolecular interactions to cell function; and to augment the Engineering Biology approach with robotics, automation and AI. Students will obtain advanced skills in programming and engineering; implement biological design across scales; place research in the context of both basic and applied science; and become cognisant of challenges such as process development and scale-up in biotechnology. Students will undertake both group and individual projects before starting their doctoral project. The EngBioCDT will take advantage of the expertise provided by the two Universities and our industrial partners, which will all be catalysts for inter-University and inter-sector training and research. Students will also have superb opportunities to engage with leading international academics, for example through an annual Summer School, and by participating in international conferences and workshops. The environment is exceptional. Bristol hosted BrisSynBio, one of six UKRI-funded Synthetic Biology Research Centres, and now hosts the Bristol BioDesign Institute and the Bristol Centre for Engineering Biology; the CDT Director is a EPSRC Fellow. Oxford, which led the SynBioCDT, received three fellowships and a programme grant in Engineering Biology, and offers vibrant translational opportunities. The applicants provide expertise in graduate training and many of them have previously worked together effectively. Our pool of >70 supervisors reflects the truly multidisciplinary nature of Engineering Biology, and includes internationally renowned researchers.