Deconvoluting the mode of action of a suite of novel antileishmanials

Leishmania spp protozoan parasites are responsible for the Neglected Tropical Disease leishmaniasis, affecting over 12 million people worldwide, with more than 1 billion at risk. With an increasing number of cases in an ever wider geographical area, and rising antimicrobial resistance, the battle against has been described as a 'losing one'. Current therapies are limited, expensive, difficult to administer and not widely accessible. Working in partnership, GSK has identified many new promising antileishmanials. To develop these further, we need to understand how they work (the Mode of Action; MoA). In this project we will using biological and biophysical technologies to uncover the mode of action of 17 hits selected for their efficacy, non-toxic profile and physical-chemical properties:
Work Package 1: Laboratory induction of antimicrobial resistance has long been observed and Next Generation Sequencing (NGS) now allows the relatively rapid identification of associated mutations in Leishmania species. Leishmania (New World L. mexicana and Old World L. major) resistance to the GSK compounds (Month 1 to 12) will be induced by in vitro evolution via stepwise increase in compound concentrations. Whole genome sequencing and evaluation of the Mode of Resistance (MoR, Month 12 to 18) using available NGS and analytical platforms will allow mutations associated with resistance will be evaluated. Validation of MoR by using gene editing technology by mutation reversal and restoration of compound sensitivity will then complete the Work Package (Month 18-36).
Work Package 2: Compounds with induced MoR mutations which have been genetically validated will then enter stage 2, where further validation will be undertaken using mass spectrometry approaches. Such technologies are now well established for the characterization of the response of cells to chemical stress and are widely utilised in the deconvolution of drug MoA, including in the laboratories of the Newcastle supervisor, Prof Matthias Trost. With Trost, leading edge mass spectrometry-based cellular thermal shift assay (MS-CeTSA; thermal proteomics) will be employed to further deconvolute the MoA of the triaged compounds (Month 24-36). MS-CeTSA allows changes in protein thermal stability associated with ligand (e.g. drug) interaction to be identified and quantified, thus facilitating the identification of drug targets in live cells (Miettinen et al, 2018). These data will be added and compared to that generated above.
Work Package 3: Compound targets fully validated in Work Packages 1 and 2 will be assessed at GSK for 'druggability', triaged and formatted into assay platforms for future HTS campaigns. In parallel, GSK medicinal chemistry expertise will assess the compounds directed at these targets for entry into hit-to-lead programmes (Month 36-42).
Write up: Month 42-48.

The CDT has five primary beneficiaries:
The CDT cohort
Our students will receive an innovative training experience making them highly employable and equipping them with the necessary knowledge and skillset in science and enterprise to become future innovators and leaders. The potential for careers in the field is substantial and students graduating from the CDT will be sought after by employers. The Life Sciences Industrial strategy states that nearly half of businesses cite a shortage of graduates as an issue in their ability to recruit talent. Collectively, the industrial partners directly involved in the co-creation of the proposal have identified recruitment needs over the next decade that already significantly exceed the output of the CDT cohort.
Life science industries
The cohort will make a vital contribution to the UK life sciences industry, filling the skills gap in this vital part of the economy and providing a talented workforce, able to instantly focus on industry relevant challenges. Through co-creation, industrial partners have shaped the training of future employees. Additional experience in management and entrepreneurship, as well as peer-to-peer activities and the beginning of a professional network provided by the cohort programme will enable graduates to become future leaders. Through direct involvement in the CDT and an ongoing programme of dissemination, stakeholders will benefit from the research and continue to contribute to its evolution. Instrument manufacturers will gain new applications for their technologies, pharmaceutical and biotech companies will gain new opportunities for drug discovery projects through new insight into disease and new methods and techniques.
Health and Society
Research outputs will ultimately benefit healthcare providers and patients in relevant areas, such as cancer, ageing and infection. Pathways to such impact are provided by involvement of industrial partners specialising in translational research and enabling networks such as the Northern Health Science Alliance, the First for Pharma group and the NHS, who will all be partners. Moreover, graduates of the CDT will provide future healthcare solutions throughout their careers in pharmaceuticals, biotechnology, contract research industries and academia.
UK economy
The cohort will contribute to growth in the life sciences industry, providing innovations that will be the vehicle for economic growth. Nationally, the Life Sciences Industrial Strategy Health Advanced Research Programme seeks to create two entirely new industries in the field over the next ten years. Regionally, medicines research is a central tenet of the Northern Powerhouse Strategy. The CDT will create new opportunities for the local life sciences sector, Inspiration for these new industries will come from researchers with an insight into both molecular and life sciences as evidenced by notable successes in the recent past. For example, the advent of Antibody Drug Conjugates and Proteolysis Targeting Chimeras arose from interdisciplinary research in this area, predominantly in the USA and have led to significant wealth and job creation. Providing a cohort of insightful, innovative and entrepreneurial scientists will help to ensure the UK remains at the forefront of future developments, in line with the aim of the Industrial Strategy of building a country confident, outward looking and fit for the future.
Institutions
Both host institutions will benefit hugely from hosting the CDT. The enhancement to the research culture provided by the presence of a diverse and international cohort of talented students will be beneficial to all researchers allied to the theme areas of the programme, who will also benefit from attending many of the scientific and networking events. The programme will further strengthen the existing scientific and cultural links between Newcastle and Durham and will provide a vehicle for new collaborative research.