Elements of a Vesicle Machine

Lead Research Organisation: Imperial College London
Department Name: Dept of Chemistry

Abstract

Despite cells being thought of as the smallest unit of life, they are actually made up smaller components that give them the ability to perform functions such as self-repair, and produce and harness energy. These components consists of large collections of proteins and lipids, the building blocks of cells, that actually work together to create higher level functions. A useful analogy is that of a city and a cell. A city often requires a wall (an outer lipid membrane), power plants (mitochondria from animal cells, and chloroplasts from plant cells) and factories and engineering firms to repair the city infrastructure (lipid homeostatic machinery). Each of these units is run by workers and machinery (lipids and proteins).In this project, we aim to take apart cells, but not at the level of the workers , instead at the level of the city infrastructure and bring them back together in a different combination with a view to constructing a new city (novel self-assembling micron scale machinery). These components, that we will be 'hi-jacking' from cells will be housed within artificial lipid vesicles that will provide a housing which mimics the ability of cells to ring-fence and protect their infrastructure. It is not yet known which components can be coupled in this manner and this is something that we aim to explore.

Planned Impact

Our goal is to explore the feasibility of generating hybrid, self-sustaining microscale machinery (HSSMMs) using an approach which is distinct from any other grouping. We will combine and manipulate gross elements of cellular machinery including mitochondrial organelles, lipid biosynthetic machinery and ion channel couples into lipid vesicles. We aim to demonstrate that these elements are isolatable and can constitute systems in their own right that can be coupled to create systems that are paradoxically more than the sum of the whole. Whilst replication is fundamental to the property of any living cell, here we choose to disregard this aspect and concentrate solely upon the ability of the HSSMMs to act as power sources that can be coupled to external systems and adapt to changes in their external environment. We believe that our high risk strategy of commandeering isolated components of the cell machinery and quantifying the interdependence of these modules will have wide ranging implications in the academic community and beyond. It would represent a novel approach, distinct to that of Synthetic Biology. In the short-term (12-18 months), the cross-disciplinary approach that is vital to tackling this strategy will bring together researchers within and thereby strengthen the cross-disciplinary networks at Imperial College, in particular the Chemical Biology Centre (CBC) and the Membrane Biophysics Platform (MBP). These feasibilities studies and their continued long term exploitation (24 months-10 years) will stimulate involvement in disciplines of chemistry, engineering, physics, mathematics, biophysics and biology. In addition to the standard methods of dissemination, meeting conferences and peer reviewed publications, we will make extensive of the MBP online Sharepoint facility and continue to organise 'Ideas Workshops'. The latter two have proven to be excellent methods of dissemination and already link over 60 research groups. In the longer term (10-15 years), HSSMMs capable of using light or mitochondrial organelles to generate ATP, which can be used as an energy currency, have the potential to revolutionise power sources for Micro-Electrical-Mechanical Systems (MEMs). By demonstrating the feasibility of first generation HSSMMs we will unlock the potential for more complex systems that can underpin novel in-vivo delivery and biosensor applications in the healthcare sector. Whilst these applications are somewhat in the future, it is vital that we capture the new IP and identify killer applications by engaging early on with the industrial sector, both during and after the feasibility studies. Our patent strategy through IC Innovations, will allow us to protect this new IP before engaging the community via established routes such the Technology Strategy Board Knowledge Transfer Networks and the CBC Technology Showcase events. The project and its deliverables sit within three of the priority research themes of the EPSRC (Nanoscience Through Engineering to Application; Energy; Towards Next Generation Healthcare). The two early careers researchers, the key resource for these studies will acquire extensive experience of systems integration and technology transfer, transferable skills that are vital to the continued future success of the academic, biotechnology and nanotechnology sectors. This exciting project is likely to stimulate increase public awareness of nanoscience. We will capitalise upon this by commissioning audio-visual content associated with our feasibility studies (e.g. recording of conference lectures or school lectures); these will then be uploaded onto facilities such as iTunesU and YouTube so that the general public can freely download this material.

Publications

10 25 50
 
Description Our feasibility studies on the hybrid, self-sustaining microscale machinery (HSSMM) project have allowed us to demonstrate that it is possible to produce HSSMMs that can move, produce and or release a cargo in response to external stimuli. This has been achieved by hijacking and manipulating component parts of a living cell's machinery and making them function in simplified contexts within HSSMMs. This set of experiments have established an exciting portfolio of potential technological applications for these micromachines including chemical synthesis, chemical sensing and molecular delivery.

These pilot studies will make a significant contribution to innovation in the UK industry, with a 10-15 year timeframe for commercial realisation and have strengthened cross-disciplinary networks across the UK bringing together 7 research groups at the Institute of Chemical Biology at Imperial College and the University of Bristol. Our results have underpinned: (i) a successful bid for an EPSRC Programme grant and Frontier Engineering grant (ii) numerous publications (iii) a Royal Society International Meeting where we presented our feasibility roadmap (v) >10 seminars including numerous presentations to industry and (vi) a CDT studentship with Kings College. The PDRAs on the project have benefitted from multi-disciplinary research training and we expect them to be future research leaders as exemplified by 4 follow-on fellowship applications submitted to build upon their contribution to this project.
Research highlights include constructing HSSMMs with the ability to:
1. drive a rotary engine embedded within the containment capsule of a HSSMM. This motor has the potential to drive a micro-propeller that we have produced and that is capable of generating thrust. This engine is driven by ATP (Adenosine-5'- triphosphate). We also demonstrated that in the future it should be possible to regenerate this ATP fuel source using chloroplasts and mitochondria as bio-inspired batteries transplanted from living cells. In the coming months we aim to couple these "batteries" with the molecular rotary engine.
2. release a pre-packaged cargo in response to a specific external stimulus. This microdelivery device couples a membrane based containment capsule with two molecular machines (A and B). The first (A) modifies the mechanical properties of the containment capsule by altering its chemical composition without compromising structural integrity. The second module (B), embedded within the membrane consists of a channel which can open or close depending on the mechanical properties of the membrane which it can sense. Following the action of module A, module B opens thereby releasing an onboard cargo. During these experiments we have been able to show for the first time that by harnessing nanoscale energetics within membranes, we can drive information flow between molecular machines without the requirement of direct contact. This system is now being translated to a microfluidic device with a view to manufacturing an insulin delivery system.
3. manufacture an onboard cargo in response to sensing the presence of an external small molecule. This HSSMM contains a linear DNA genome with the genetic information required to produce a coloured gene product after detecting the presence of a specific small molecule, in this case a sugar. This HSSMM mimics the ability of living cells to adapt their response to the environment by altering gene expression. We have also started the development of HSSMMs that couple cargo production with light and evolve over generations.

In addition, we have developed high-throughput platforms for generating 2-D and 3-D microfluidic droplet interface bilayer (DIBs) networks. These can be used to build up complex HSSMM systems and are now being exploited in two follow-on projects with commercial collaborators. These technologies breakthrough downstream enabled us to develop microfluidic strategies for manufacturing compartmentalised vesicles following the project which thereby tackled a long standing bottleneck in the biotech industry.
Exploitation Route This programme will make a significant contribution to innovation in UK industry, with a 5-15 year timeframe for commercial realisation. Our early stage proof of concept studies on HSSMM technologies have demonstrated that they have the potential to drive new strategies and approaches to drug molecule-membrane screening (1-5 years), drug delivery (5 years), drug release (5 years), in-situ bio-diagnostics and sensors (5-10 years), small molecule detection/tracking (5-10 years) and the biological microreactors. The latter application is in advanced PoC stages of testing.
Sectors Agriculture, Food and Drink,Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description Collaboration with pharmaceutical industry looking at use of artificial cell technologies for nanomedical applications
First Year Of Impact 2016
Sector Chemicals,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Societal,Economic

 
Description EPSRC CDT PhD Studentship (Engineering communication in artificial cells)
Amount £95,000 (GBP)
Funding ID 1373739 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2012 
End 09/2016
 
Description EPSRC CDT PhD Studentship: Drug-membrane interaction screens
Amount £71,400 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2011 
End 09/2014
 
Description EPSRC DTG Studentship (awarded through the Department of Chemistry): Optical Traps+DIBs
Amount £80,000 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2012 
End 09/2016
 
Description EPSRC Industrial CASE Studentship
Amount £67,000 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2012 
End 09/2016
 
Description EPSRC Pathways to Impact Funding
Amount £74,400 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2011 
End 06/2012
 
Description EPSRC Programme Grant (Sculpting Dynamic Amphiphilic Surfaces), EP/J017566/1
Amount £4,772,920 (GBP)
Funding ID EP/J017566/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 06/2012 
End 05/2017
 
Description Frontier Manufacturing: Scaling Up Synthetic Biology (EP/K038648/1)
Amount £5,160,000 (GBP)
Funding ID EP/K038648/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2013 
End 10/2018
 
Description Joint collaboration: Protocell technologies for Healthcare
Amount £150,000 (GBP)
Organisation AstraZeneca 
Sector Private
Country United Kingdom
Start 10/2016 
End 09/2020
 
Description Marie Curie Initial Training Network: NextGenAgriChem
Amount £3,260,000 (GBP)
Organisation European Research Council (ERC) 
Sector Public
Country Belgium
Start 03/2014 
End 03/2018
 
Description Collaboration with Proctor and Gamble 
Organisation Procter & Gamble
Country United States 
Sector Private 
PI Contribution Collaboration with Proctor and Gamble (supported by CASE studentship) EPSRC CASE Studentship starting in October 2012. We are also in discussions with P and G with a view to funding an associated PDRA as well.
Start Year 2012
 
Description Collaboration with Syngenta 
Organisation Syngenta International AG
Department Syngenta Ltd (Bracknell)
Country United Kingdom 
Sector Private 
PI Contribution Development of DIB platform
Collaborator Contribution Exploitation of DIB platform for studying membrane translocation in plant systems.
Impact Collaboration just started, exploring potential of the technology: multi-disciplinary activity.
Start Year 2013
 
Title Membrane based protein-protein communication 
Description We have demonstrated for the first time that it is possible to engineer protein-protein communication between proteins, without the need for direct contact by using biological membranes as the information highway. This technology is now being further developed to manufacture artificial cells/organelles and will help to unlock our understanding of protein-protein communication networks in cells. 
Type Of Technology New/Improved Technique/Technology 
Year Produced 2011 
Impact Has underpinned the development of a new generation of bottom-up artificial cells capable of sensing their environment. It has in effect introduced a new engineering component into the design of protocells 
 
Title Microfluidic generation of 2-D and 3-D droplet interface bilayer networks 
Description Using microfludic platforms we can now manufacture droplet interface bilayer networks using the contours of microfluidic chips to control the shape of the networks whilst also using the delivery flow rate and droplet size to influence droplet packing geometries with the host chambers. These new approaches allow individual droplet position and composition to be controlled, paving the way for complex on-chip funtional network synthesis and bespoke scaffolds for artificial cells, organelles and hybrid self-sustaining micromachinery. Industrial applications are currently being pursued. 
Type Of Technology New/Improved Technique/Technology 
 
Description International Microfluidics Congress 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact International conference: disseminate findings of research and look for new end users of the technology
Year(s) Of Engagement Activity 2015
 
Description Presentation to the general public (Friends of Imperial College) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Presentation aimed at addressing public concerning associated with manufacturing artificial cells and manipulating biological systems.
Year(s) Of Engagement Activity 2015
 
Description Technology showcase presentation to industry (AstraZeneca) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Workshop and associated talk aimed at translating EPSRC breakthrough to industry.
Year(s) Of Engagement Activity 2015