Cystic fibrosis (CF) is caused by inherited mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) and is one of the most widespread inherited genetic disorders. Although the disease affects multiple organs, the primary cause of morbidity and mortality is cystic fibrosis related lung disease. Due to its recessive nature, i.e. requirement for two defective copies of the gene, and easy accessibility of the lungs to inhaled gene therapy agents; CF is an excellent candidate for gene replacement therapy. With funding from CF Canada, our lab has been developing gene therapy vectors to “functionally cure” individuals with CF. To achieve this goal, we have genetically engineered baculovirus, a harmless insect virus, to permanently insert genes of interest into a safe location within the human genome using two different cutting-edge genome-editing technologies (TALENs and CRISPR-Cas9). We have also modified the baculovirus genome so that it can “hide” from the immune system long enough to permit site-specific integration of therapeutic genes to occur. By engineering baculovirus to deliver all the “tools” required to insert a functional copy of the CFTR gene into the genome of lung cells, we hope to be able to provide a safe, economical and effective treatment for CF patients irrespective of their CFTR mutation status, gender or genetic background. Additionally, these studies would validate this platform as a general gene therapy vector for broader gene therapy applications in tissues other than the lung, as well as ex vivo correction of human stem cells.
Alpha-1-antitrypsin deficiency (AATD) is a monogenetic disorder commonly associated with adult onset lung diseases, including emphysema, chronic obstructive pulmonary disease (COPD) and airway inflammation and affects an estimated 190 million people worldwide. A promising treatment approach for AATD is gene therapy. The idea behind gene therapy is to provide an individual who has defective copies of the AAT gene, with a functional, good copy of the gene. Recently great strides have been made in gene therapy, however none of the current approaches can permanently deliver a good copy of the AAT gene into the human genome in a controlled manner. Our lab is currently employing the use of adeno-associated virus (AAV) and baculovirus gene therapy vectors and two different cutting-edge genome editing technologies (TALENs and CRISPR-Cas9) to insert a functional copy of the AAT gene, permanently, into a safe location within the human genome.
- Dr. Theo Moraes, Sick Kids