Clinical grade cell separation technologies in the Newcastle Cellular Therapies Facility

CONTEXT
The production of cells as medicines is a regulated process and can only be conducted in accordance with manufacturing procedures, quality control processes, and in licensed premises according to EU Directives. Newcastle University and Newcastle upon Tyne NHS Foundation Trust have invested in the necessary infrastructure to build, maintain and license production facilities compliant with Good Manufacturing Practice (GMP). The necessary licenses for manufacturing cells as medicines have been obtained, and with the Clinical Trials Unit at the Royal Victoria Infirmary and the Institute for Transplantation at the Freeman Hospital, clinical support for patients is enabled by the provision of specific beds for cell therapy patients. The Newcastle Cellular Therapies Facility is currently running two clinical trials in cellular therapies. The first is the MRC Autologous Limbal Stem Cell trial for eye trauma with Professor Francisco Figuieredo, and the second is the tolerogenic dendritic cell (TolDC) trial for rheumatoid arthritis with Professor John Isaacs. In addition, HTA approval has been granted for autologous pancreatic islet cell therapy with Professor Stephen White. Furthermore, Professor Dickinson is Director of the Haematopoietic Stem Cell Transplant laboratory which is currently responsible for releasing approximately 400 cell therapy products per year to adult and paediatric patients suffering from malignant blood disorders and immune deficiency. A number of qualified staff with practical, scientific and regulatory expertise are employed within the NCTF who are able to initiate and develop products for clinical trials in cellular therapies. The NCTF is subjected to inspections by the MHRA, HTA and internal audits. The staff employed include a Pharmacy Qualified Person, a Quality Assurance Manager, and Health Professional Council-registered Biomedical Scientists who work closely with clinicians and clinical trial teams.

AIMS AND OBJECTIVES
We seek support through the MRC Regenerative Medicine Capital Funding scheme for the purchase of equipment to be housed within the production GMP cleanrooms that have the capacity to purify populations of cells by their expression of cell surface markers. Antibodies conjugated to magnetic beads that permit the selection or depletion of chosen cell types is established technology. Miltenyi-Biotec produces two different machines, or platforms, that can exploit the magnetic bead technology to purify cell populations. The first is the Clinimacs Plus that uses magnetic beads but is designed to purify chosen cells into sterile bags that can then be issued ready for infusion or further manipulation. The second is the new Clinimacs Prodigy that offers a closed workstation capable of processing and purifying and culturing cells in a programmable way and for which specific processes can be designed and optimised. It is state-of-the-art technology that can be used for new process development as well as clinical grade manufacturing of cell therapies.

APPLICATIONS & BENEFITS
The Clinimacs Plus would support the TolDC trial. The Clinimacs Prodigy will find immediate use in development of new protocols, exploiting our collaboration with the Hannover group led by Professor Eva Weissinger, for virus-specific T-cell therapy in patients who contract CMV after allogeneic bone marrow infusions. Protocols designed by Miltenyi-Biotec render the Prodigy appropriate and feasible for this research. The equipment requested will therefore give NCTF the capacity to aid current clinical trials and research into new clinical trials, and develop new protocols in cell therapy to meet the MRC strategy. In addition, research, teaching and training in cell therapy manufacture will be supported across the EU by hosting this equipment in our facility as Professor Dickinson participates in EU funded initiatives to establish standards in cell therapy to support consistency across multi-centre trials.

OBJECTIVES
Research and development for current and future clinical trials of which Newcastle University is either a sponsor or a partner is our objective. Cell purification by positive or negative selection, or combinations of the two approaches, will be developed in clinical trials of dendritic cell vaccines, and viral-specific T cells for CMV infection, in the first instance. These are technologies that can be extended to multiple indications. For example, viral-specific T cells could be used to treat EBV infection and deployed in solid organ transplant recipients. Partnerships are being developed with trial sponsors, industrial partners, and through academic and applied research projects to exploit the capacity of NCTF to manufacture cellular therapies in a GMP licensed facility.

THE CLINIMACS PRODIGY: The instrument permits automated, standardized cell manufacturing processes in a closed system, and is equipped with a cell processing unit for washing and fractionating cells by centrifugation, image recognition to detect macroscopic layers of cell product, and an integral cell separation unit, based on magnetic bead cell-selection techniques. The device also possesses an integrated cell culture chamber. Input ports allow for the sterile removal and replenishment of media, whilst cells can be monitored using an integrated microscope. The device therefore is a tool for the design and optimisation of GMP-Grade processes and provides a means of producing cell-specific products for human use.

CLINIMACS PLUS: is an automated cell separation system based on Miltenyi-Biotec magnetic bead separation technology. It enables the operator to perform clinical-scale magnetic enrichment of target cells. Processes using this technology will be validated and made available to other clinical trials. This capability will permit manufacture of cellular therapies for users beyond Newcastle and support the MRC's strategic aims for the UK development of cellular therapies

BENEFICIARIES OF THIS CAPITAL INVESTMENT

Firstly, the availability of this technology will enhance research and development capacity in NCTF by resolving issues of limited equipment and support faster progression of the ARUK funded TolDC trial. The progression of trials in rheumatoid arthritis has the potential to positively impact upon large patient populations.

Secondly, the acquisition of this technology will permit the development of protocols for the efficient isolation and expansion of viral-specific T-cell therapies - initially for the treatment of CMV positive paediatric SCID patients. From the development of appropriate methods we would extend T-cell specific therapy to adults, and in solid-organ transplant recipients and across other viral infections such as EBV. The development and dissemination of protocols using these technologies will have impact across the EU through the EU funded AGORA program for which Professor Dickinson and Dr De Havilland are partners, as well as the established collaboration with Hannover in the development of the clinical use of viral-specific T cells after stem cell transplantation.

Thirdly, the capacity to purify cells will, in the future, facilitate pre-clinical research and clinical trials in other indications where differentiated cells may de deployed. We are already in advanced discussions with trial sponsors in indications as diverse as stroke and bone fractures to add to our current portfolio in blindness, arthritis, diabetes and leukaemia. Future trials in these indications also have the potential to benefit large patient populations. This will be especially so as allogeneic tissue banks of cells differentiated for specific purposes are developed.

Fourthly, we are engaged in major funded cell therapy initiatives in rheumatoid arthritis research and in the development of methodologies to manufacture IMPs and ATMPs. The validation of methods that can be disseminated through collaboration, peer-review, or marketing materials, will support the standardisation of methods in cellular therapies within the EU. A network is already established for which Professor Dickinson is the co-ordinator and therefore we have a process by which international collaboration can be exploited for the development and distribution of optimised methods in cell therapies.

Fifthly, it is the establishment of this technology within our licenced GMP facility that is a unique alliance. This provides the UK with a strategic capability for research and exploitation by other institutions and clinical trial sponsors.The collection of expertise in these emerging methods will enable policy makers to take advantage of developments in Newcastle to provide a template for future research and commercial-scale manufacturing processes by industrial partners.

In terms of timescale the equipment we request can be provided within 6-8 weeks of order and could therefore be in the current clinical trials. Within 6-12 months we would anticipate having developed validated protocols for viral-specific T cell manufacture. Over the next 2-5 years Newcastle intends to build on its success in developing cellular therapies and extend its clinical trial portfolio, and be able to offer development of purified cells as a specialised service for research into cellular therapies, including those differentiated from pluripotent stem cells.

Newcastle bases its research and manufacturing capacity on HPC-registered and qualified biomedical scientists and a dedicated research staff infrastructure.The equipment will facilitate staff training in research methods of cell purification technologies, in development and validation of GMP compliant protocols, and in the development of manufacturing, quality control, quality assurance and product release processes for cellular therapies. A set of core skills will be gained for the UK as a whole, which is critical in order to lead development in these emerging technologies.