Project: Cultured Autologous Oral Mucosal Epithelial Cell Sheet

CAOMECS is being developed for the treatment of patients with total limbal stem cell deficiency (LSCD) with moderate or severe symptoms caused by exogenous trauma or endogenous eye disease. The CO objective is to restore the epithelium on ocular surfaces of total LSCD patients. The corneal epithelium is a tissue with a dynamic turnover. Its renewal is achieved though the presence of the limbal stem cells. Failure of this process can result in a variety of serious and intractable disorders of the ocular surface including loss of corneal transparency with reduction in vision and blindness. Surgical treatments for total LSCD include amniotic membrane grafts combined with limbal stem cell grafts which have not undergone tissue culture, and, where necessary, subsequent corneal grafting. Tissue engineering, allowing the development of cell sheets from autologous epithelium may serve as a suitable alternative methods for cornea repair. Much progress has been made in the field of corneal tissue engineering in recent years. If cells can be harvested from either eye of a patient with cornea damage or a very small quantity (1-2mm) of the the patient's non-affected eye's cornea can be cultured to re-establish the epithelial sheet that could be used for repair of the corneal epithelial in the damaged eye. Where this is not possible then the use of cadaveric tissue or living related donors must be considered. This causes further complications associated with the use of immunosuppresive drugs to prevent rejection. Consequently, there is an urgent medical need to provide safer, more stable and effective medicinal products for the treatment of total LSCD patients which must consider other autologous epithelial cells such as oral mucosa from the patient. _x000D__x000D_CellSeed has a significant programme of research in the area of regenerative medicine associated with cornea repair. We have developed proprietary technology that allows us to harvest autologous cells from the patients and grow these externally for clinical replacement of the epithelial layer of the damaged cornea. _x000D__x000D_Our technology is based on the use of Poly N-isopropylacrylamide. This is a unique polymer that exhibits thermally reversible soluble-insoluble changes in aqueous solution in response to temperature change across the lower critical solution temperature (LCST) of 32 degrees centigrade. The polymer chains of acrylamide hydrate to expand in water below the LCST, while the isopropyl group dehydrating to form compact, insoluble conformations above the LCST. _x000D__x000D_When this temperature responsive polymer is coated onto a cell culturing plastic plate using bionano interface technology, the surface of the plate demonstrates reversible hydrophilic-hydrophobic response depending on the temperature change across the LCST of 32 degrees centigrade. This surface becomes hydrophobic over 32 degrees centrigrade which enables cells to attach to the surface and grow._x000D__x000D_However, when the temperature is reduced to as low as 20 degrees centigrade, the polymer surface becomes hydrophilic and the hydrated polymer chains cause the cultured cells to detach easily in a single cell sheet with intact cell-cell interactions. This is an important aspect for the use of these cell sheets for clinical therapy. The cells retain the function of fibronection and the cells are not damaged by the culturing or detachment process. The significant clinical benefit of this approach is that since cell damaging enzymes are not required the cell sheet is highly effective when transplanted to patients due to the retention of the communication between the cells. Furthermore, CAOMECS does not require suturing onto the cornea and so avoids inflammation due to the sutures._x000D__x000D_This technology has direct application to the culture of cell sheets that can be used for repair of damaged corneal epithelium. The technology has the ability to be used with a range of cell types including autologous and allogeneic._x000D__x000D_Our technology has been evaluated in vitro and in vivo using animal models of cornea damage with significant success. We are now in the position to enter the next stage of development for this product which is the formal clinical evaluation of the technology in patients with cornea damage. In preparation for this work we have had extensive discussion with the European Medicines Agency (EMEA) regarding the nature, performance and validation of the clinical trial._x000D__x000D_Furthermore, this platform technology has additional uses within the clinical environment and also for tissue engineering and regenerative medicine more widely. One of the significant benefits of the polymer coated plates is the ability to construct more complex tissues within the culture plate. This allows the construction of more complex 3 dimensional multiple cell layers which could be as scaffolds for regenerative medicine.

Acronym	CAOMECS (Reference Number: 5221)
Duration	04/01/2010 - 04/07/2012
Project Topic	Ocular trauma or disease may lead to severe corneal opacification and consequently severe loss of vision as a result of complete loss of corneal epithelial cells. We are developing a novel cell sheet technology to provide a new treatment for patients with limbal stem cell deficiency.
Project Results (after finalisation)	As cooperation P in the EUROSTAR PROJECT #5221 CAOMECS, the Department of Ophthalmology of the University Hospital Salzburg was involved in clinical and molecularbiological evaluation of the cell-sheet technology, method validation, process validation and also transport validation. Furthermore clinical trial application as well as the approval to conduct the clinical trial has been submitted and accepted by the austrian regulatories (ethics commitee). _x000D_CO results:_x000D_1) Evaluation and optimization of oral tissue biopsy, including educational video for all participating clinical centers _x000D_2) Evaluation and optimization of biopsy transport for cell-sheet cultivation and transplantation_x000D_3) Establishment of molecular biological analysis of cultivated cell-sheet_x000D_4) Approval to conduct clinical trial by austrian regulatories_x000D_5) Development of clinical trial protocols, manuals and SOPs_x000D_6) Inclusion of five patients for the clinical trial, that has been postponed due to financial problems of the CO P
Network	Eurostars
Call	Eurostars Cut-Off 3

Project partner

Number	Name	Role	Country
4	CellSeed Europe S.A.R.L.	Coordinator	France
4	Gemeinnuetzige Salzburger Landeskliniken Betriebsgesellschaft mbH, Landeskrankenhaus Salzburg, Universitaetsklinik für Augenheilkunde und Optometrie	Partner	Austria
4	Genesis Pharma S.A.	Partner	Greece
4	Hospices Civils de Lyon	Partner	France