Lentiviral gene therapy rescues p47phox chronic granulomatous disease and the ability to fight Salmonella infection in mice

• Published in: Gene therapy 2020-09, Vol.27 (9), p.459-469
• Main Authors: Schejtman, Andrea, Aragão-Filho, Walmir Cutrim, Clare, Simon, Zinicola, Marta, Weisser, Maren, Burns, Siobhan O., Booth, Claire, Gaspar, Hubert B., Thomas, David C., Condino-Neto, Antonio, Thrasher, Adrian J., Santilli, Giorgia
• Format: Article
• Language: English
• Subjects: 13; 13/106; 13/31; 14; 38; 38/77; 42; 631/1647/2300/1850; 631/250/249; 631/532/1542; 82; Biomedical and Life Sciences; Biomedicine; Brief Communication; Cathepsin G; Cell Biology; Chronic granulomatous disease; Chronic infection; Gene Expression; Gene Therapy; Hematopoietic stem cells; Human Genetics; Immunodeficiency; Mutation; NAD(P)H oxidase; Nanotechnology; Phagocytes; Primary immunodeficiencies; Salmonella; Sepsis; Stem cell transplantation; Stem cells
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/s41434-020-0164-6

Chronic granulomatous disease (CGD) is an inherited primary immunodeficiency disorder characterised by recurrent and often life-threatening infections and hyperinflammation. It is caused by defects of the phagocytic NADPH oxidase, a multicomponent enzyme system responsible for effective pathogen killing. A phase I/II clinical trial of lentiviral gene therapy is underway for the most common form of CGD, X-linked, caused by mutations in the gp91 phox subunit of the NADPH oxidase. We propose to use a similar strategy to tackle p47 phox -deficient CGD, caused by mutations in NCF1 , which encodes the p47 phox cytosolic component of the enzymatic complex. We generated a pCCLCHIM-p47 phox lentiviral vector, containing the chimeric Cathepsin G / FES myeloid promoter and a codon-optimised version of the human NCF1 cDNA. Here we show that transduction with the pCCLCHIM-p47 phox vector efficiently restores p47 phox expression and biochemical NADPH oxidase function in p47 phox -deficient human and murine cells. We also tested the ability of our gene therapy approach to control infection by challenging p47 phox -null mice with Salmonella Typhimurium, a leading cause of sepsis in CGD patients, and found that mice reconstituted with lentivirus-transduced hematopoietic stem cells had a reduced bacterial load compared with untreated mice. Overall, our results potentially support the clinical development of a gene therapy approach using the pCCLCHIM-p47 phox vector.