Generation of a site directed gene integration platform for induced pluripotent stem cell lines.

To understand how the human brain works and therefore how to intervene when it goes wrong we need to be able to study brain cells such as neurons. Living human neurons are virtually impossible to obtain in order to understand how basic cellular processes occur in the human brain. Stem cells offer the ability to produce human neurons and other brain cells to use in such studies. Recently there has been a revolution in the field of stem cell research. Now it is possible to take skin cells from disease patients, transform them into stem cells, and then from these generate patient specific neuronal networks in culture.

The ability to insert novel genes into neurons is of great importance and allows scientists to study cell behaviour in response to different triggers such as gene mutations, but also allows them to control the activity of cells. However this is particularly difficult in cells such as neurons, as they do not divide, and often requires specialised reagents such as viruses, which are used as vectors to insert genes into non-dividing cells. When using viruses or other common insertion methods, these genes are often inserted randomly into the cells own DNA and if inserted in the wrong area, can alter the ability of the cell to carry out its normal functions. One solution is to modify the genome of the stem cell to incorporate a region of DNA that can serve as a 'docking site' or 'launch pad' for novel genes that is in a safe area. In this way novel genes will always be incorporated into the same place in the cells genome without altering the cells normal functions. As a stem cell can be made to differentiate into many different types of cell, creation of a stem cell with a docking site for gene insertion will provide a platform for researchers to assess the function of inserted genes in many cell types.

In this project we propose to combine the skills of Axol biosciences, a company who specialises in providing stem cell derived neurons to all researchers with the skills of the Neural stem cell group at AU to create such a platform stem cell line. The platform line will be tested by the insertion of genes encoding Channelrhodopsin (ChR) proteins, which will make the neurons sensitive to laser light, allowing researchers to non-invasively stimulate neurons into the types of activity that are observed in the brain. This is a technically difficult challenge and will develop the skills of both partners. Scientists from Axol will also be trained within Dr Rhein Parri's lab in techniques that will enable them to test their cell lines to ensure that they behave as cells do within the brain. This work will provide highly specialised scientists with the unique combination of skills in both fields that will enhance their research capabilities. The experience of both interchangers in both academic and commercial environments will build new skills within each organisation that will be embedded in future research and development to produce future leaders in this area of research.

The number of people over the age of 65 in the UK has now risen to over 10 million, with a concomitant rise in age related disease and care costs, dementia care costs more than that of cancer and heart disease combined. Basic research in neuroscience has the capacity to improve public health, by providing new targets and models to test and improve pharmaceutical interventions for the treatment of neurodegenerative disorders. Development of technologies to facilitate such research, along with the training and knowledge gained in this FLIP will have far reaching benefits to a wide group of beneficiaries.

Who will benefit?

AU, Axol Biosciences, Neuroscience/research community, Pharma and Biotech companies, Economy, Students, NHS Budgets, Government, patients and their carers, The Public.

How will they benefit?

Academics/Researchers
Aston and its collaborators will immediately benefit from the training and skills acquired, along with access to the platform line. Other researchers will then have access to the quality controlled platform line (2 yrs) providing a tailored gene expression system to easily study proteins/genes of interest in cutting edge systems. This will reduce start up times for new projects and remove the technical challenges which preclude many researchers from the adoption of such technologies.

Axol Bioscience
Within year one of the grant Axol will benefit though acquisition of electrophysiology skills to enhance their QC procedures, and research. They will also benefit through development of a unique product line (2 yrs) which has been developed in conjunction with users and is amenable to the needs of most researchers. The line will also be compatible with other cell lineages as Axol grow and diversify their product base (3-10 yrs). They will also benefit through an enhanced understanding of their customer base.

Pharma and biotech companies
The adoption, of a commercially available neuronal model capable of selective gene expression will have a knock-on effect to other SME's. Functional neuronal models and reporter cell lines will require cell based reagents as well as neurophysiology platforms such as MEAs, imaging platforms and microscopes. Hence increasing demand for such products benefiting SMEs supplying them.

NHS
Growing numbers of elderly people increase the pressure on NHS budgets. Dementia costs the UK economy £23 billion a year. This FLIP will provide an easily manipulatable iPSC platform line and complex neuronal models amenable to controlled gene expression. This will provide a powerful tool to facilitate the scientific discoveries to inform the development of new treatments aimed at ameliorating neurological disease (5-15 yrs).

Students
Within the lifetime of the grant (1-2 yrs), practical examples of such cutting edge technologies, can be incorporated into undergraduate and MSc lectures at AU. Producing graduates prepared for their future careers by ensuring they are informed of the latest scientific developments, and inspiring students at a crucial stage to consider a career in brain research.

Government
This project will highlight the benefit of investing in high quality research and training that can provide longer term economic benefits through both industrial progress (3-5 yrs) and healthcare benefits (5-15 yrs).

Public
There is growing public interest in the functioning of the brain and the use of stem cells. Disseminating the findings of this research (1-2 yrs) in line with our existing commitment to public engagement will increase public understanding and appreciation of such cutting edge research.

Economy
In the medium to long term (3-5 yrs) this project has great potential to positively impact the UK economy, increasing the global economic competitiveness of a vibrant UK based start-up company, and ultimately providing a resource for UK based scientific discovery.