Optically-Guided Nanoparticles and Cell Scalextrics

Lead Research Organisation: University of Nottingham
Department Name: Sch of Pharmacy

Abstract

Steering nanoparticle transport in human cells - why is this important?
Viruses seem to travel effortlessly into tissues and cells, being transported selectively in the body to reach the sites where they can cause most harm. They do this by breaching cellular barriers such as the outer or plasma membrane of cells and use human cellular machinery to make copies of themselves. Drug molecules on the other hand spread non-selectively throughout the body thus reducing effects where they are most needed, and causing adverse side-effects where they are not wanted. One reason for this is that current synthetic carriers for drugs and diagnostic agents, unlike viruses, are unable to effectively cross biological barriers and then reach specific sites inside cells. An artificial particle that could transport through tissue, in a manner analogous to a virus and then into defined cell locations but without causing disease, would therefore revolutionise healthcare applications. This would be particularly important for the early stage diagnostics and therapeutics needed in developing nations and for ageing populations.

What do novel polymer-coated gold nanoparticles have to offer?
These materials are an optimal test platform for proof-of-concept studies described in this application. Firstly, gold particles can be tracked in cells using a number of microscopic approaches including the newly developed highly sensitive four-wave mixing imaging system available to this team. They can be coated with a variety of polymeric materials that will help to guide them into cells and into specific cell locations. We have previously shown that polymers which are capped with functional 'keys' to enter natural cell portals can have their entry switched on and off by small increases in temperature which cause them to change their conformations. We also have shown that we can generate these temperature increases at gold nanoparticles inside cells through laser pulses but without damaging the cells. By attaching the temperature-responsive polymers to gold, it should be possible to use laser pulses to switch the functional keys on and off, and in this way guide particles to reach defined cellular locations. This will help to unravel the mechanisms by which materials travel in cells, thus enabling us to guide diagnostics and therapeutics to where they are required.

Impact.
A major hurdle to effective therapy against major disease burdens such as cancer, coronary heart disease and neurodegeneration is our inability to direct therapeutic molecules such as genes and proteins to specific tissue and defined compartments inside cells. This is a major objective of this application and progress here could have widespread implications for academia, industry and the society that they serve. One could envisage a commercial application in which a combined imaging and guiding instrument (e,g, ultrasonic probes and imaging) is used with a set of nanoparticles with functionality for specific disease markers, with a potential for truly selective personalised therapies.
Better diagnostics are also needed that allow earlier detection of disease and thus better healthcare outcomes. Successful completion of this work could allow development of an imaging/detection platform where specific markers of disease could be detected through their interaction with selective receptors on gold nanoparticles guided to intracellular sites by the local laser-heating method. When it is considered that 1 in 3 individuals in the EU will be affected directly or indirectly by cancer by 2010, it is clear that earlier detection and intervention will bring marked benefits to patients, carers and society as a whole.
Longer-term development could generate impact through a new biomedical technology i.e. laser-guided therapeutics wherein local heating by focused ultrasound guides biodegradable responsive nanoparticles in humans.

Planned Impact

Successful development of materials and instrumentation to guide particles in cells would be a world-leading advance, with many potential applications. Further refinement in future grants would pave the way to bioresponsive self-reporting/self-healing biosensors and therapeutics, and represent a revolutionary advance in healthcare,

Benefits to industry
Probes based on precise placement of diagnostic particles and with the ability to expose in a switchable manner e.g. a nucleic acid sequence, could lead to a family of optogenetic sensors. Nature Methods selected optogenetics as breakthrough of the year (2010) & there is great scope for IP in this area. 'Cornerstone' patents in site-directed diagnostics could bring industry benefits through the formation of a spin-out or, more probably, through licensing of technology. Via links with Eminate and BioCity Nottingham, we have ready access to SMEs operating in sensors/diagnostics (e.g. CellAura). For therapeutics, switchable intracellular exposure of anti-sense ligands would enable selective knock-down e.g. of oncogenes. Our collaborations with leading pharma (AstraZeneca, GSK, MedImmune) facilitate routes to licensing and commercial exploitation of drug delivery systems. If preliminary data is promising, we will file patents and apply for Follow-On funding to develop the technology in collaboration with industry partners. There is potential further industry benefit through training of both PDRAs in multidisciplinary work, but also PhDs in our groups, who will be very suitable for high technology SME/pharma jobs as a result of interactions in the project. It is estimated that~ 48 % of pharmaceuticals employees are linked to or engaged directly in R+D, and each employee contributes to sales worth £233,000 per person (ABPI, 2007): even small numbers of people working in this area can have a very high economic impact. Recent changes in the industry have led to a re-focus to more advanced medical technologies, based on the ealisation that biomolecule and cell-derived therapies are a potential £5 billion market ("New Industry, New Jobs". BIS, 2010). This grant is the first step leading to a technology platform for these future applications.

Benefits to policy makers
Better diagnostics and more effective therapeutics are needed in developing nations and for ageing populations. The increasing emphasis on preventative medicine to reduce healthcare budgets demands earlier diagnosis. For therapeutics, poor targeting leads to inefficient use, undesired side effects and greater medical intervention, with burdens on patients, carer populations and healthcare budgets. Estimates of healthcare costs are difficult to predict, but the projected worldwide $199 bn drug delivery market for 2016 suggests this is an area where spend will be increased. Policymakers will benefit more if they invest in highly innovative delivery technologies now. Further benefits should accrue though enhanced visibility for EPSRC-funded science in this area. The UK is still regarded as an attractive base for pharma R+D but the restructuring of the industry could lead to erosion of this position - by funding high profile work in the pharmaceutical area, EPSRC is sending a strong signal of support to the industry. Indeed, this is reflected in recent EPSRC Priority Areas, (Techniques for biomedical understanding, Diagnostics, Therapeutic Technologies and Medicines) into which this proposal fits closely.

Benefits to the public
Better diagnosis aids 'wellness' while better delivery aids cure. Technologies developed from controlled particle transport could solve the current targeting problems for many diagnostic (e.g. MRI contrast enhancement agents) and drugs. AuNPs have been demonstrated for anticancer efficacy in vitro (JACS. 2010, 132, 1517) but extension to guided NPs in vivo would bring benefit in the longer-term through earlier indications of medical problems and better treatments.

Publications

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Kakde D (2016) Synthesis, characterization and evaluation of in vitro toxicity in hepatocytes of linear polyesters with varied aromatic and aliphatic co-monomers. in Journal of controlled release : official journal of the Controlled Release Society

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Moody PR (2015) Receptor Crosslinking: A General Method to Trigger Internalization and Lysosomal Targeting of Therapeutic Receptor:Ligand Complexes. in Molecular therapy : the journal of the American Society of Gene Therapy

 
Description The grant is now complete and some initial key findings have been published. (Moody, P. R.; Sayers, E. J.; Magnusson, J. P.; Alexander, C.; Borri, P.; Watson, P. and Jones, A. T. Receptor crosslinking - A General Method to Trigger Internalisation and Lysosomal Targeting of Therapeutic Receptor:Ligand Complexes. Molecular Therapy 2015, 23 (12), 1888-1898.)
The work showed that certain therapeutic molecules can be directed to the parts of a cell where they are most needed, and that for the breast cancer antibody Trastuzamab, this can lead to better efficacy in cell lines.
A second significant paper was published in 2018 (Bioconjugate Chem., 2018, 29 (4), pp 1030-1046 DOI: 10.1021/acs.bioconjchem.7b00704) which demonstrated that polymer-decorated nanoparticles can be targeted to specific cell entry points, and that this process can be switched on or off by small temperature change-induced variations in polymer architecture. However, the data also showed that the nanoparticles are 're-routed' inside the cells, which also has important implications for intracellular therapeutic targeting.
Exploitation Route New drug delivery mechanisms and better therapies for difficult cancer targets
Sectors Chemicals,Education,Healthcare,Pharmaceuticals and Medical Biotechnology

URL http://www.nature.com/mt/journal/v23/n12/abs/mt2015178a.html
 
Description Findings have been used to date to enhance our collaboration between Cardiff, Padova and Nottingham. Further press has led to significant interest http://www.cardiff.ac.uk/news/view/185481-pharmacy-research-makes-headlines The paper from this project in 2015 contributed to my selection to peer review for the European Union Nanocharacterisation Laboratory (EU-NCL), which involves guiding academics and spin-outs in the route to translation of investigational healthcare materials. In addition, a full paper from this work has now been accepted at Bioconjugate Chemistry and will appear online shortly
First Year Of Impact 2015
Sector Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Cultural,Economic

 
Description Radiotherapy activated materials for enhanced cancer treatments
Amount £539,154 (GBP)
Funding ID EP/N03371X/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 12/2016 
End 11/2018
 
Description Wellcome Trust Institutional Strategic Support Fund: - Collaboration Panel: Cross-Disciplinary Award
Amount £29,552 (GBP)
Organisation Wellcome Trust 
Department Wellcome Trust Institutional Strategic Support Fund
Sector Charity/Non Profit
Country United Kingdom
Start 09/2017 
End 02/2018
 
Description Astra Zeneca Project 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution New collaboration with funding from AstraZeneca starting starting in April 2016 running to June 2017
Collaborator Contribution The partners will synthesise targetted drug delivery vectors for Cardiff University to test in in vitro models Have now submitted joint Cardiff University - AstraZeneca BBSRC-IPA application. Expected oucome June 2017
Impact None to date as side fro presenting some data slides at conferences
Start Year 2016
 
Description University of Padova 
Organisation University of Padova
Country Italy 
Sector Academic/University 
PI Contribution We have hosted a number of Masters and PhD students for research stays. We have provided materials, training in polymer synthesis, access to instrumentation and facilities.
Collaborator Contribution Excellent Masters students, of which 6 have stayed on to carry out PhD studies in Nottingham. The Padova group have also hosted our PhD students and PDRAs, giving hem training in pharmaceutical formulation science and pharmacokinetics.
Impact Multiple papers (> 10) EU/EPSRC Grants (Value > 500,000 GBP) Multidisciplinary (pharmacy, pharmacology, chemistry)
Start Year 2006
 
Description National Eisteddfod of Wales Meifod 2015 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Through EPSRC funding (EP/J021334/1) a large science exhibition was organised under the title of Working Polymers; This was held over a eight day period at the Science and Technology Pavilion of the National Eisteddfod of Wales - Meifod Mid-Wales- August 1 - August 8 and officially opened by the First Minister for Wales Carwyn Jones.
The aim of this exhibit was to highlight the science of polymers and the fact that we are now totally dependant on them but to also bring introduce EPSRC funded research in the ATJ laboratory on the use of polymers for designing nanomedicines to target diseases such as cancer. Approximately 130,000 visited the Eisteddfod over this eight day period and >20,000 were counted into the Science Pavilion. The Exhibit was staffed by Cardiff University academics and undergraduate students. Public engagement training was provided for all activity staff by Professor Jones.
Feedback was extremely positive and there was much discussion surrounding the various polymer activities in display e.g. surface of rugby balls, nappies, cardiac stents with controlled release of drugs. This also attracted the media and numerous interviews were conducted with BBC Radio and TV and also S4C. A major outcome of these is that the public now clearly view science as a fundamental part of culture and this is why the Science and Technology Pavilion sits proudly and comfortably at the heart of this very large cultural festival.
Year(s) Of Engagement Activity 2015
URL https://blogs.cardiff.ac.uk/pharmacy-engagement/2015/10/22/national-eisteddfod-2015/