School of Molecular Biosciences

Orthobioactive coatings (HealiOst)

Professors Manuel Salmerón-Sánchez and Matt Dalby have developed a novel bioactive polymer coating that can be applied to complex orthopaedic implant geometries using plasma polymerisation. The coating causes biologically correct organisation of the cell adherence protein fibronectin. As fibronectin is adsorbed it unfolds into a fibrillar confirmation forming biological networks and exposes the RGD cell adhesion domain and also the growth factor binding site in close proximity. This allows growth factors such as BMP2 to be bound to the growth factor site at very low yet highly efficient doses for stimulation of cells in the environment. BMP2 is currently widely used clinically but of great concern as it is administered at supraphysiological doses to achieve effect. Our technology allows for biologically safe doses to provide a greater and more targeted effect. The full coating system was used clinically for the first time in 2017 with the successful treatment of Eva the dog, a veterinary patient from the Glasgow Small Animal Hospital. Being subject to a severe fracture following a car accident, Eva was at the stage of having her leg amputated when HealiOst was deployed in a last effort to repair the bone and save her leg. Eva’s veterinary surgeon, Mr William Marshall, used bone graft coated with HealiOst and BMP-2 to fill the 2cm bone gap with the result being full repair of the bone within 6 weeks of implantation. The team have secured ERC funding to treat further veterinary cases and are pursuing human use of this technology through funding provided by Find a Better Way – a landmine charity focussed on helping the survivors of landmine blast injuries. A patent has been filed for HealiOst and collaborations with partner organisations are focussed on clinical translation for human orthopaedic indications.

NanoKick bioreactor

Professors Matt Dalby and Stuart Reid (University of Strathclyde) have developed the Nanokick bioreactor, currently subject to patent application. It uses tiny vibrational movements to mechanically stimulate mesenchymal stem cells to form bone in vitro. Unlike other bioreactors it uses standard cell culture consumables (e.g. 6 well plates and flasks) making it easy to integrate into standard cell culture protocols. Further, it requires no complex media formulations / supplements to achieve osteogenesis – Nanokicking alone is enough to consistently stimulate osteogenesis in 2D or 3D culture. The team are currently working with partners including the Scottish Blood Transfusion Service to use Nanokicking to manufacture an off the shelf injectable cell therapy for bone repair as part of their Find a Better Way funded project aiming to help landmine survivors. The need for clinically implantable bone cells arises from bone being the second most grafted tissue behind blood and existing products / techniques having limitations in supply or regenerative potential. Data from in vivo studies suggests that Nanokicked cells are more effective at bone repair than standard mesenchymal stem cells due to their phenotypic priming to form osteoblasts. As part of the project the team are progressing towards first-in-human trial of the cell therapy with funding already secured from Find a Better Way.

Using Drosophila for kidney disease therapeutics

Professor Julian Dow is at the forefront of research in ‘omics, ion transport and kidney function in the genetic model Drosophila melanogaster (1,2). This has shown commonality of kidney function between humans and Drosophila, allowing the modelling of kidney disease using Drosophila, with the aim of therapeutics development. The group has now developed a successful model for kidney stones in the fly ‘kidney’ (3, 4) with funding from National Institutes of Health, USA as well as BBSRC Sparking Impact funds that enabled development of in vivo screening of kidney stone formation using the fly kidney.

The key advantage of the Drosophila system is that it permits rapid, reproducible formation of stones in an intact, transparent tissue,so allowing informative analysis and compound screens to be performed for the first time. He and Professor Shireen Davies are also partners in an EU-funded H2020 Marie Curie Innovative Training Network on renal development and disease, using Drosophila to model other kidney diseases including Inborn Errors of Metabolism.

References
1 Proc. Natl Acad.Sci. USA 111, 14301-14306 (2014).
2 Nat Commun 6, 6800, doi:10.1038 ncomms7800 ncomms7800 .(2015).
3 Am. J. Physiol.-Renal Physiology 303, F1555-F1562 (2012).
4 J. Urol., doi:S0022-5347(13)03645-8 (2013).

Stress tolerance in crops

Researchers in the group of Professor Anna Amtmann have discovered a protein in the chromatin of plants that attenuates gene induction under salt or water stress, and enhances biomass. Before publishing their results in The Plant Cell [1] they discussed their discovery with Matthew Hannah, project leader at Bayer CropScience, Gent, Belgium.

Following independent testing of the transgenic lines in the Bayer laboratories, a licence agreement was achieved between Bayer and UoG, and a common patent application was filed. Bayer provided bridging funds for the post-doctoral researcher at Glasgow during the negotiations, and they committed to a £0.5 M Industrial Partnership Award from the BBSRC. The 3-year project on the molecular and physiological functions of the gene is now well underway.

Professor Amtmann describes the experience as very positive. “The scientific feedback we are getting from Dr Hannah is well-informed and constructive, and it is exciting for us to follow the progress of our gene in the crop improvement programs at Bayer. It was hard work and sometimes nerve-wrecking to get to this stage but it in the end it is a net gain for all partners involved. Clearly, fundamental science and commercialisation can go hand-in-hand.”

[1] Perrella et al. (2013) Plant Cell 25: 3491-3505. doi:10.1105/tpc.113.114835

Developing greener ways of controlling pest insects

Professor Shireen Davies is currently leading the 14 -partner EU-funded nEUROSTRESSPEP consortium to develop novel and greener insecticidal agents based on insect neuropeptide mimetics. Neuropeptides are regulators of critical life processes in insects and comprise new insecticidal agents to selectively reduce the fitness of pest insects, whilst minimising detrimental environmental impacts. The project nEUROSTRESSPEP (http://www.neurostresspep.eu/home) has developed insecticidal prototypes based on ‘omics, physiology and chemistry, as well as genetic pest management strategies. 

Optimising antibody production in CHO cells by engineering protein folding and secretion

One of the major challenges facing the biopharmaceutical industry is to produce sufficient therapeutic proteins in a cost effective way to meet market demand. Over the past twenty years great progress has been made in both upstream and downstream processing which has enabled some products to reach the market place. To improve the level of protein production from mammalian cells we need to consider not only the expression levels of the protein but also the capacity of the cell to fold assemble and secrete recombinant proteins. The folding, assembly and secretion of proteins from mammalian cells has been a major focus for the laboratory of Professor Neil Bulleid. UCB Pharma funded two PhD students to try and increase the yield of therapeutic antibodies. This provided opportunity to relate fundamental research to the production of biopharmaceuticals. Their work in the Bulleid lab showed how engineering antibodies can have adverse effects on assembly (1) and allowed details of stress pathways to be elucidated (2). The company benefited from access to the knowledge base and by being engaged in projects that would be too risky for them to fund internally.

1. Stoyle, C. L., Stephens, P. E., Humphreys, D. P., Heywood, S., Cain, K., and Bulleid, N. J. (2017) Biochem J. 474, 3179-3188
2. Chalmers, F., van Lith, M., Sweeney, B., Cain, K., and Bulleid, N. J. (2017) Wellcome Open Res. 2, 36

The Protein and Nucleic Acid Characterisation Facility (PNACF)

The Protein and Nucleic Acid Characterisation Facility (PNACF), managed by Dr Sharon Kelly, offers a wide variety of biophysical techniques, including fluorescence, circular dichroism, UV-VIS, FTIR, isothermal calorimetry, and surface plasmon resonance (SPR), to support Research & Development by analysing structural integrity, stability, and interactions with other macromolecules, ligands, and membranes. The PNACF can provide a biophysical characterisation service to test the integrity of bio-therapeutics during drug development. Extensive characterisation underpins all product development activities and therefore must be done thoroughly to fulfil FDA and ICH guidelines (ICH Q6B).

One company we worked with were required by ICH guidelines to demonstrate the integrity and stability of their protein therapeutic drug using the recommended biophysical characterization procedures described above. Different batches of a protein therapeutic drug were tested and compared with reference drug using Far and Near UV Circular Dichroism and Fluorescence studies to ensure structural integrity and stability.

It was found that batches older than 6 months gave spectra which were dissimilar from freshly prepared reference therapeutic drugs (prepared under identical procedures). Older batches were prone to aggregation and were less thermally stable than freshly prepared therapeutic.

The results of the stability trial allowed the Company’s Research and Development team to alter the formulation of the therapeutic to improve stability.

Targetting receptors for short chain fatty acids to treat and metabolic and inflammatory diseases

In recent years Professor Graeme Milligan has been exploring ways in which mimicking the health beneficial effects of short chain fatty acids that are produced in high levels by the microflora could be applied to the production of ‘functional foods’, via either pre-biotic or pro-biotic strategies. In parallel with this effort Professor Milligan has been studying the underlying biology of the receptors that are activated by these receptors and whether they might be novel and tractable targets for the treatment of metabolic dysfunctions such as type II diabetes.

Publications stemming from this research have attracted great interest from both large and medium sized pharmaceutical companies. Professor Milligan was invited to travel to a number of companies to give presentations and discuss potential collaborations.

The most satisfactory potential partner in terms of providing each of potential funding, beneficial non-overlapping expertise and a history of previous links between Professor Milligan’s group and scientists employed by the company, was Astra-Zeneca.

Following discussions, Astra-Zeneca supported an Industrial Partnership Award (IPA) application led by Professor Milligan to BBSRC. This provides a total of £1.5 Million in funding over a 4 year period and Astra-Zeneca provides £150K in cash. The company has also provided a range of non-commercially available ligands and in return will gain pre-publication insights into which of the receptors for short chain fatty acids is most suitable to target as well as a number of animal models that will be generated within the project.

Provision of sensory neuronal cells to the pharma industry

The Centre for the Cellular Microenvironment was funded by the NC3Rs to address the DRGNet challenge as part of their CRACK IT open innovation programme. The aim of the DRGNet challenge was to identify sustainable sources of sensory neurons isolated from human dorsal root ganglia (hDRG). Pharma raised the challenge as recently several drugs for pain, that had been successful in animal testing, subsequently failed phase 1 human trials. Access to a local, affordable and reliable source of hDRG sensory neurons may enable the pharmaceutical industry to identify and discard early in development compounds destined to fail.

We formed a very capable team with expertise in cell isolation, maintenance and characterisation that was led by, from Glasgow, Prof Andrew Hart, Dr Mathis Riehle, David I Hughes and Mair Crouch, and Gareth Miles from St Andrews, alongside collaboration with the industrial partners Grünenthal GmbH (Aachen, D) and Metrion (Cambridge, UK). After ethics application, the ability to isolate the hDRG cells was established. The hDRG neurons were shown by several partners, including industry, to respond as expected to capsaicin, pH and ATP. The hDRG neurons show a similar distribution of immunological subtype markers as other mammals. Several different action potential response types, typical I/V curves and TTX insensitive sodium currents were regularly recorded allowing to fully characterise the usefulness of these cells for in vitro testing. We are currently in an early phase of our exit strategy for DRGNet Scotland.

Site-specific recombination for rapid, efficient DNA rearrangements in Synthetic Biology

Professor Marshall Stark and Dr Sean Colloms are leaders in the field of site-specific recombination which was recognized in the award of a £4.3M 5-year ‘sLoLa’ grant by BBSRC to the University of Glasgow (P.I. Professor Stark), for development of a platform for rapid and precise DNA module rearrangements in Synthetic Biology, using sitespecific recombination. Our principal industrial partner for this project is Ingenza Ltd. The aim of our research with Ingenza is to develop novel recombinase-based tools and systems for assembly of metabolic pathway genes and regulatory components, and subsequently manipulate the assemblies to optimize metabolite production. The grant runs from 2013-2018, so the experimental programme is still at an early stage. However, we are already investigating promising approaches, both in vitro and in industrially useful producer organisms. The research will enhance Ingenza’s portfolio of methodologies for systematic genome modification and thereby support the interests of their customers. Demonstration of utility of our methods in an industrial context will also fulfil an explicit aim of the BBSRC funded research programme, will increase the publishability of the research, and may lead to valuable intellectual property.