Work package 6
Validation of Gene Transfer System by Assessment of Safety in Vitro and in Animal Model
The overall objective of this work package is to assess safety of the transposon mediated gene delivery system which will be used in the clinical trial to transfer the gene sequence of the therapeutic PEDF protein (Pigment Epithelium Derived Factor) to the patient’s pigment epithelial cells.
One safety analysis of the transfected human iris pigment epithelial (IPE) and retinal pigment epithelial (RPE) cells represent the determination of the genomic integration profiles of the applied transposon construct with the recombinant PEDF gene sequence. The in vitro transfected pigment epithelial cells will be analysed by using the method of LAM-PCR/deep sequencing.
Afterwards homologous IPE and RPE cells of two different normal pigmented animal models will be transplanted subconjunctivally in healthy mice (C57/BL6) and subretinally in rabbits (Chinchilla Bastard) (Fig. 2).
In mice the genomic safety will be examined using large-scale integration site analysis by comparison of non-transplanted with transplanted tissue to proof whether expansion or a clonal imbalance exists in the graft. In the rabbit model it will be determined the genotoxicity by demonstrating of the biodistribution, trafficking, localisation and persistence of genetically modified cells in vivo.
Completion of the studies in animal models will initiate the final review process for the clinical trials by the regulatory authority and will include Milestone 3 (M3), the preparation of data for inclusion into the investigational medicinal product dossier (work package 8).