Gene therapeutic intervention during ex vivo lung perfusion as a platform to modulate human lung disease

Lung transplantation (LTx) is the ultimate therapy to treat patients suffering from for end-stage lung disease. Unfortunately, LTx is accompanied by a low survival rate. Primary Graft Dysfunction (PGD) is a severe form of acute lung injury after LTx that is a major cause of poor survival. Although much progress has been made in understanding the risks and mechanisms of PGD, there is still no effective treatment. Ex-vivo lung perfusion (EVLP) maintains the isolated lung grafts in a physiologic and metabolic state before implantation and provides a unique platform to offer treatment interventions to prevent PGD. With a CRISPR-based genome editing strategy, we aim to study the potential of silencing PGD-related genes (e.g., IL-6) during EVLP. In this PhD research, two distinct delivery vehicles will be examined in their ability to efficiently but transiently, introduce the CRISPR-Cas9 machinery into the cells of the lung: adeno-associated viral (rAAV) vectors and Virus-Like Particles (VLPs). After in vitro optimization of delivery, through reporter gene expression, we aim to investigate if CRISPR-Cas9 gene knockout (targeted against a PGD-gene of interest) can reduce the severity of PGD both in vitro and in vivo.