Research projects

Gene editing approaches are being developed as a novel therapeutic strategy for genetic diseases, having already reached the clinic. The aim of this project is the development of CRIPSR editing therapeutics to correct frequent pathogenic variants in genes responsible for the rare neurometabolic diseases phenylketonuria (PKU) and propionic acidemia (PA), using cellular and/or animal models previously generated. Both PKU and PA are autosomal recessive disorders resulting from defects in the liver enzyme phenylalanine hydroxylase (PAH) or in propionyl Coa carboxylase (PCC), respectively, that cause the toxic built-up of metabolites. Current dietary and medical treatments do not prevent cardiac and neurologic alterations in PA, or are associated with suboptimal outcomes in PKU, highlighting the high unmet clinical need of novel corrective treatments. 

PAH variant c.1066-11G>A is the second most frequent pathogenic variant causing PKU, causing a splice defect that results in a functionally null protein. In this project we will test adenine base editors (ABE) to correct the variant, using a knock in (KI) HepG2 model and a humanized mouse model with the variant. In PA, for the frequent PCCB variant c.1218_1231delinsTAGAGCACAGGA we will use Prime editing in mutant stably transformed fibroblasts and/or iPSC. Finally, using a KI HepG2 model with a pseudoexon activating variant (PCCA c.1285-1416A>G) causing PA, we will test pairs of gRNAs to excise the intronic pseudoexon. Gene editing efficiency and functional rescue will be monitored via NGS, transcript, protein and activity analyses, to provide the proof of concept of an innovative, one-time corrective therapy approach for these diseases.