MICA: Development of Metrics and Quality Standards for Scale up of Human Pluripotent Stem Cells

Many diseases remain untreatable in spite of advances in medicine world-wide. In recent years there has been much interest in using human pluripotent stem cells (hPSCs), unspecialised cells which can be coaxed using molecules that are found normally in the developing body to produce any cell type. Such stem cells can therefore be used to produce particular repair tissues for patients with different diseases that are so far incurable or for damaged tissues following injury. In order for this to be possible in a manner affordable to our healthcare systems, we need to generate banks of stem cells of different human background which have different immunological tissue types. These can then be used to make desired specialised cells tailored to the patient. However at the moment we do not have reliable ways to grow the cells in large numbers without damaging them, or changing their properties so that they cannot make healthy tissues. In this proposal we will develop the conditions for generating healthy stem cells in large numbers while protecting them from damaging influences. We will generate a system that will be suitable for an automated cell factory so that the cells can be monitored and maintained in pristine stress-free conditions and expanded in a stable way enabling them to in turn generate healthy tissues for the clinic. Such a system has been developed by a company, Tokyo Electron, which now has a base in the UK. We will evaluate a whole series of different properties of these hPSCs and work out which attributes of the cells predict their health and retention of the ability to make more stem cells and form specialised cells. These properies will be compared against commercial and other alternative hPSC test systems and, in the second phase, fed back to further improve culture conditions. In this subsequent second phase we will trial the improved culture conditions with continued cell monitoring for 'good' and 'bad' cell markers in the automated cell factory. The established validated properties will be made available to other researchers so that they can use them to improve their expansion of PSCs and subsequently generate different specialised cell types for different patient needs.

Expansion and banking of "off the shelf" allogeneic human pluripotent stem cell (hPSC)-derived lineages poses a realistic option for tissue repair, affordable to public/private health care systems. However, the regenerative medicine community currently lacks affordable and scalable systems. This bottleneck for translational programmes requires novel solutions as a matter of urgency, with the availability of clinical grade human embryonic stem cell, and in the near future human induced pluripotent stem cell lines. The overarching goal of this proposal is the development and qualification of a novel system for the automated scale up of hPSCs invented by Tokyo Electron (EU base in Stevenage), for subsequent differentiation into therapeutically valuable lineages. The novel, entirely enclosed, automated culture system suitable for next generation stem cell therapy manufacturing will use non-invasive image evaluation and non invasive biochemical/genetic monitoring to allow evaluation of cell health and phenotype during cell expansion. Thus, only healthy cells can be selected for continued culture or entry into therapeutic differentiation protocols. In this proposal we will develop the metrics to ensure optimal conditions for stable expanded hPSC culture, including growth rates/phenotype and karyotypic/epigenetic normality. Deep phenotyping will include cell morphology via refinement of computer algorithms, genetic (e.g.CGH array) and epigenetic profiling and both discovery and targeted secretome metabolite/proteome analysis. Pluripotency will be assured via development of high throughput novel differentiation marker analysis replacing teratoma formation. We will generate new biomarkers based on culture medium analysis (secretome, biosensor data) to refine culture conditions and generate new media/substrate combinations. We will deliver online/Cloud-based metrics analysis tools available to UK/ other stem cell communities adaptable for other scale up applications.

Tissue degeneration in an aging or sick population presents an ever-increasing clinical burden for patients and healthcare systems, with costs rising rapidly. The idea that stem cell, including hPSC, products might relieve this burden is tempered by the lack of scalable culture and differentiation systems that allow cell therapies to be delivered quickly at affordable cost. Through partnership with an inwardly-investing company, Tokyo Electron Ltd (TEL), our 2 phase proposal will address this need by combining deep hPSC analysis with state of the art bioinformatics and engineering approaches to develop and validate minimum sets of biomarkers that quantifiably define cell quality standards. This will be of broad use to hPSP user communities, and will also enable TEL's development of a fully enclosed Smart Cell System that can scale up and bank quality controlled hPSCs for differentiation to valuable lineages.
Researchers in the UK regenerative medicine community will benefit from Open Access to optimised protocols and reagents for culturing, monitoring and quantifying quality criteria of hPSCs. Novel resources, knowledge and tools (e.g. biosensors) will be produced from bioinformatics analyses of deep phenotyping data derived by cell characterisation, proteomics, lipidomics, metabolomics, genetic and epigenetic analyses. These will underpin and impact, multiple UK and international initiatives: £40m Regenerative Medicine & Capital Equipment Call funded 5 RCUK and 3 BHF Platform hubs; 3 MRC clinical grade hESC derivation centres; £13m MRC-Wellcome hiPSC scheme; euro29m IMI EU Bank for hiPSC (EBISC). Researchers in these initiatives and elsewhere will benefit from the new analytical kits and biosensors that spin-out from the industrial partnerships developed. Close connection (see letter of support) and researcher exchange and training within our consortium and between these initiatives will foster the next generation of interdisciplinary scientists.
Biopharma and Industry will immediately benefit via £2.9m of inward investment from TEL into its Stevenage site and £500K cash/in-kind to this project. Consortium partners have funding and/or partnerships with Intercytex, Syngenta, GSK, AstraZeneca, Roche, Clyde Biosciences & Pluriomics. As well as partnering with the academics in this consortium, TEL is linking with UK companies such as Orla Protein (Newcastle; culture substrates), Cell Guidance Systems (Cambridge; media) and Plexus Inc. (Edinburgh; robotics). These companies will benefit from our consortium, as will those we approach for the development of quality control testing kits/assays (e.g. Stem Cell Technologies, Miltenyi Biotech, LifeTech) and matrices (e.g. Nunc, Corning and Greiner). Exploitation, IP and licencing will be via Tech Transfer Offices and TEL; see Pathways to Impact and MICA for details.
The NHS will directly link to the proposal since Manchester Academic Health Science Centre (MAHSC) is one of only five Department of Health designated AHSCs in the UK, which recognises excellence across research, innovation, education, patient service and translational medicine. Brison is employed within Central Manchester NHS Foundation Trust (CMFT), a key partner of MAHSC, wherein CMFT's remit is to translate basic research into clinical practice working with professional bodies e.g. RCPath. These routes will ensure impact in the healthcare system.
The public will benefit by being the recipients of new healthcare technologies in the future. They will be well informed since the academic partners have proven track records in engagement through channels ranging from television, radio and newspapers to public engagement days, science cafes, school education events and websites (e.g. www.eurostemcell.org).
Regulators will be kept abreast of developments through existing links via e.g. HFEA Scientific Advisory Committee (Brison & Young), MHRA (Brison), and stem cell banking and regulation (Stacey).