Base and Prime Editing Strategies for Recessive Dystrophic Epidermolysis Bullosa
Lay summary
Individuals with recessive dystrophic epidermolysis bullosa (RDEB) suffer from widespread blistering of external and internal skin, along with complications such as poor wound healing, narrowing of the oesophagus (food pipe) due to scarring, corneal erosions (damage to the outer surface of the eye), fusion of fingers and toes, and the development of skin cancer. The condition is caused by mutations in COL7A1, the gene encoding type VII protein, and there is currently no specific treatment.
The goal of our research is to provide a treatment that will improve the quality of life of patients by preventing and/or treating skin complications of RDEB. The project aims to develop a strategy that could be used to treat a significant subset of RDEB patients carrying selected mutations.
The project will investigate the feasibility of base and prime editing strategies in primary RDEB cells in vitro and ex vivo, but also in vivo using new delivery methods such as lipid nanoparticles or virus-like particles which are devoid of viral genetic material. The project will target 10 COL7A1 recurrent mutations which are present in 13.5% of the RDEB patient population.
Scientific Summary
This project aims to develop base and prime editing strategies to correct recurrent COL7A1 mutations in primary keratinocytes, fibroblasts and iPSCs from RDEB patients.
We will design the most efficient gRNAs and pegRNAs in combination with base and/or prime editors (BEs; PEs) which will be delivered into cultured RDEB cells using either mRNA nucleofection, virus like particles (VLPs) or lipid nanoparticles (LNPs).
Subsequently, LNPs or VLPs will be delivered by intradermal or intravenous injections into nude mice grafted with RDEB skin equivalents. Restoration of type VII collagen function will first be assessed in skin organoids, then in skin equivalents grafted onto nude mice.