This proposal seeks to cement the scientific collaboration further between Prof. Jim Tour and Dr. Robert Pal. Our combined research aim is to develop and test small molecular agents that once activated by low dosage of Ultra-violet light can selectively destroy cancerous cells and tissues. This collaboration has been established, when Prof. Tour visited Durham as the 2015 Durham Lecturer at our department. Prof. Tour proposed a, yet unanswered, scientific challenge as part of one of his lectures, that due to my research expertise and available instrumentation I was able to propose a series of experiments to answer with. Exactly one year and lots of hard work later now we are in the fortunate position that our collective findings have been submitted to Nature and is currently under review of publication. As a result of this well-oiled collaboration not only this mutually beneficial milestone have been met, but a further more extensive NIH research grant have been submitted 12/2016 to facilitate the synthesis of the next generation of complexes to be studied at Durham (decision pending). However, the above mentioned proposal would only facilitate research staff and consumables cost to be met at Rice University. Up to this date Dr. Pal as a Royal Society URF, who as a URF is severely limited to apply for funding to undertake such activity, had no associated funding to visit Prof. Tour's lab, a visit that would facilitate mutual knowledge exchange and the establishment of a suitable optical microscopy facility to conduct any required preliminary screening of active compounds before the subsequent Durham live cell evaluation. There is no better way to showcase the scientific timeliness and magnitude of our combined research area than the 2016 Nobel Prize in chemistry, that has been awarded for 'Molecular Machines', and the 2014 Nobel Chemistry Prize what has been awarded for 'Super-resolution microscopy'. This small travel grant could facilitate this and accelerate the future success of the project greatly.

There are more than one million annual amputations globally as a result of vascular diseases, trauma and cancer. Due to the increasing rate of diabetes and the population ageing, a growth of amputation is expected with the prediction that the amputee population will double by 2050. A prosthesis allows a certain restoration of functional mobility after an amputation. However, neither passive nor active prostheses can directly address the fundamental problems of chronic pain and muscle atrophy in millions of amputees worldwide. Chronic amputation-related pain impairs function. In addition, the early decline in the use of the affected limb results in progressive muscle atrophy with strength loss. Concurrently, a mechanical adaption occurs in order to compensate for the collective effects due to limb loss. A common compensation strategy is to overload the intact limbs in terms of time and intensity, which will cause secondary musculoskeletal disorders, further compromising their health-related quality of life. The project will target the clear unmet needs of amputees by developing a novel functional electrical stimulation (FES) device. A ground-breaking, computational approach based on predictive musculoskeletal modelling will be developed and integrated into the device to design patient-centred rehabilitation. This device has the potential to prescribe the optimal rehabilitation protocol in pain management and mobility enhancement; its long-term application will prevent the onset and progress of musculoskeletal disorders and ultimately improve the quality of life. Dr Ding will receive support from a multidisciplinary team of researchers, business experts, clinicians and patients to ensure the project will impact various stakeholders. Patients: given the high risk of multiple complications after an amputation, the development of such a new generation of therapies will be attractive to millions of amputees worldwide. Of further importance, are the costs of amputee rehabilitation services. To a civilian amputee, the rehabilitation cost in the first 5 years is approximately $107,200. To the military amputees, the costs will be even higher as they are younger at the time of injury and societal costs will be incurred over a longer period. As such, to prove the efficacy of the novel device in amputee rehabilitation will have a significant socioeconomic impact. Clinicians: the computational rehabilitation design has the potential to enhance clinical decision making in the rehabilitation pathway. By modelling individual patient's performance and predicting the outcomes in the in-silico environment, the modelling approach will improve patient's satisfaction to current practice. The project will create a collaborative environment for engineers and clinicians to share intellectual investment and make a broad impact of computational modelling and simulation in the clinic. Medical device industry: the application of FES therapy to the amputee population will directly expand the market size of electrical stimulators. The FES manufacturers could instantiate the rehabilitation protocol in their existing products to fulfil the application or develop a standalone product for amputees. The incorporation of the predictive amputee musculoskeletal model in the product design will lead to a reduced time to market and improved patient outcomes. The long-term FES application will also benefit the manufacturers of FES components such as electrodes, sensors and battery and add a broader socioeconomic impact. Educational beneficiaries: the project will allow Dr Ding as a STEM (Science, Technology, Engineering and Maths) Ambassador to run activities in local schools focus on the biomechanics applications of STEM subjects. Also, she will disseminate impressive animations and case studies on musculoskeletal modelling and simulation in her lab website to attract a broad range of students to study biomechanics, making a wider academic impact.

An important need for future bio-medical and fundamental cell biology is the targeted in vivo destruction of selected cells and cell types. Cancer affects every country regardless of financial status, medical infrastructure or the availability of highly-skilled specialists and is responsible for 8.2 million deaths/year worldwide. Our vision for the future is to develop a series of light-activated molecular nanomachines - Nano-drills - to target cancerous cells selectively and safely eradicate them. Currently this can only be facilitated using highly invasive surgical or radiotherapeutic procedures that are often harmful to administer. We have already demonstrated that our Nano-drills (molecular Nano-Machines, MNMs) can selectively destroy cells using small quantities of ultraviolet activation light. The next phase of our research is four-fold. First, we must enable multi-two-photon (2PE) activation on our existing microscope system using biologically safe near-infra-red (NIR) light for live-cell studies. Second, extend the family of cell type and pathological condition specific nano-drills to allow a wide range of cancer cells to study. Third, design nano-drills that can be activated using plain visible light in order to comply with already established protocols in biomedical research. Fourth, create a new breed of nano-drills that will first be internalised by the targeted cancer cells and subsequently allowing eradication to be triggered them from within. This will promote a more controlled single-cell precision research tool. Preliminary studies showed that our MNMs can be successfully activated using 2PE. In order to continue this research, we need a fast, high resolution live cell capable Multi-Photon Microscope (MPM) to image live cells. Unfortunately, due to the nature and unique instrumental requirement an off the shelf MPM system costs in the region of £500,000. However, we propose to develop a suitable custom system by adapting existing equipment at fraction of the cost. We will equip our Leica SP5 II confocal system with a tuneable high repetition rate state-of-the-art NIR laser. This experimental modification will allow us to directly compare live-cell molecular nanomachine activation with both UV and 2PE excitation and exceed the capabilities of a standard off-the-shelf microscope. We have already purchased all associated auxiliary optical components needed for attaching the NIR laser. We advanced the instrument build to a stage, where after sourcing the required laser system a simple 'plug and play' approach will yield the multi-photon upgrade on our instrument within a few weeks. Once fully developed it could form a new extremely high precision biological research tool and opens new horizons toward wound healing and tumour/cancer progression studies. It could pave the way towards a non-invasive treatment of chemotherapeutic agent resistant cancers, such as metastatic breast cancer. This project has a potential game-changing impact on a global scale.

Dynamical systems with many degrees of freedom arise in a wide range of applications, including large scale molecular dynamics, climate and weather studies, and electrical power networks. The challenge in simulation is normally to extract statistical information, for example the average propensity of a given state of the system or the average time that elapses between certain events. Simulation data is easy to generate but often poorly utilized. The goal of this project is the development of a data-driven method for the automatic detection of a simplified description of the system based on a set of collective variables which can be used within efficient statistical extraction procedures. These slowest degrees of freedom are typically the most important ones. The dynamics are characterised as fluctuations in the vicinity of given state punctuated by relatively rare events describing transitions between the states. Efficiently identifying collective variables is the crucial first step in the design of coarse-grained models which can allow many order of magnitude increases in the accessible simulation timescale. By automatically finding collective variables, we can greatly simplify rapid study and comparison of many systems. The research builds on the technique of diffusion maps, whereby the eigenfunctions of a diffusion operator are used to characterise the metastable (slowly changing) states of the system. The potential impact of automatic coarse-graining will be felt most profoundly in fields such as rational drug design, where it is necessary to select specific drug molecules for their properties in interaction with some target, e.g. a protein. Bio-molecular simulation depends on the use of very specialised and intensely developed simulation codes which are the products of many years of development and government investment. In order to accelerate the implementation and testing of novel algorithms in this important area, this project includes a detailed plan for software development within the EPSRC-funded MIST (Molecular Integrator Software Tools) platform. Testing of the software methodology will be conducted via collaborations with chemists and pharmaceutical chemists, including researchers at Rice University (Houston, Texas) and Memorial Sloan Kettering Cancer Research Center (New York).

Millions of medical devices are surgically implanted every year, with annual sales approaching US$500 billion worldwide. Failure of implanted devices designed to be permanent can be as high as 20%, impacting patients' quality of life and burdening health services. Glucose sensors are used by most diabetics in the UK, with fine needle electrodes to sense glucose in the outermost tissue - they have recommended lifetimes of only 10-14 days because foreign body encapsulation renders them inaccurate, with each disposable unit costing £50. The foreign body response (FBR) is the hostile immune cell reaction of the body to implants, with chronic inflammation, infection and fibrosis being the major underlying causes of implant failure. With sustained support from Wellcome Trust and EPSRC over the last fifteen years, including a current Large Grant, we are developing novel cell-instructive polymers to reduce and ultimately eliminating medical device failure. To underpin cell-instructive polymer development, we need to be able to monitor the response of the body to novel implants in real-time. Only a snapshot of the complex biological interplay between inflammatory pathways is provided by current histological assessment of inflammatory responses measured on explants. The lack of technology to sense real-time changes of these complex processes hampers our ability to comprehensively understand these intricate inflammatory mechanisms in the hunt for polymers providing the best implant outcomes. We propose the development of a disruptive method to achieve continuous, minimally invasive monitoring of implants in both animal models and humans. Longitudinal real-time measurements of signature inflammatory markers and FBR will be made possible using an innovative wireless bioelectronic approach: conductive nanoantennae will be decorated with antibodies to achieve continuous and minimally invasive electrical monitoring of cytokines and macrophages in a multiplexed fashion. This novel wireless monitoring method will allow us to assess new polymers in situ in real-time, aiding their successful development. When used in humans, sensing will allow the continuous monitoring of the body's response to the new implant and therefore faster and better therapies that will ultimately improve implant success, patient outcomes and savings for healthcare providers. It will have broader application in the clinic for a variety of conditions where (device-unrelated) fibrosis is the source of morbidity and mortality. People with diabetes suffer disproportionately from adverse implant reactions as well as chronic wounds. Through a clinical partnership with a diabetologist, we will develop an impedance sensor that does not require nanoantenna injection for earlier clinical adoption proved on glucose monitors worn by healthy volunteers. This proposal has been co-developed by our interdisciplinary and international team, integrating expertise in cell-instructive materials, immunology, analytic devices engineering, clinical application and medical device commercialisation. The scope spans EPSRC, MRC and BBSRC remits, making it challenging for a single council and review college to fully address the multifaceted expertise and methodological range assembled to tackle this unmet need. Benefits for the biomaterials and medical device fields include mechanistic understanding and acceleration of the novel device development process which will speed impact through MedTech products to improve options for clinicians. Immunologists will better understand the kinetics of the inflammatory response enabling more complete mechanistic descriptions. Reciprocal benefits for the rapidly advancing bioelectronics discipline will be through the clinical and pre-clinical examples it will deliver, along with the methodological experience that will be contained within the journal publications and patent filings.