Gene Therapy with pCCLchimP47 -phox Lentiviral Vector Achieves Long-term Reconstitution of NADPH Oxidase Activity in p47 -phox Chronic Granulomatous Disease (CGD) Mouse Model

Chronic granulomatous disease (CGD) is an inherited deficiency of the superoxide-generating phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, resulting in recurrent, severe bacterial and fungal infections. The X-linked form of this disorder (X-CGD) results from mutations in the X-linked gene for gp91phox, the larger subunit of the oxidase flavocytochrome b558. In this study, we used a murine model of X-CGD to examine the long-term function of retroviral vectors for expression of gp91phox based on the murine stem cell virus (MSCV) backbone. NADPH oxidase activity was reconstituted in neutrophils and macrophages for up to 18 to 24 months posttransplantation of transduced X-CGD bone marrow into lethally irradiated syngeneic X-CGD mice. Southern blot analysis and secondary transplant data showed proviral integration in multilineage repopulating cells. Although relatively small amounts of recombinant gp91phox (approximately 5% to 10% of wild-type levels) were detected in neutrophils after retroviral-mediated gene transfer, superoxide-generating activity was approximately 20% to 25% of wild-type mouse neutrophils. Expression of gp91phox is normally restricted to mature phagocytes. No obvious toxicity was observed in other hematopoietic lineages in transplant recipients, and provirus-marked cells were capable of reconstituting secondary transplant recipients, who also exhibited NADPH oxidase–positive neutrophils. MSCV-based vectors for long-term expression of gp91phox may be useful for gene therapy of human CGD targeted at hematopoietic stem cells.

Download Full-text

Long-Term High-Level Reconstitution of NADPH Oxidase Activity in Murine X-Linked Chronic Granulomatous Disease Using a Bicistronic Vector Expressing gp91phoxand a ΔLNGFR Cell Surface Marker

Human Gene Therapy ◽

10.1089/104303403321618164 ◽

2003 ◽

Vol 14 (7) ◽

pp. 651-666 ◽

Cited By ~ 18

Author(s):

Mohammed A. Sadat ◽

Nancy Pech ◽

So Saulnier ◽

Brendan A. Leroy ◽

Johann P. Hossle ◽

...

Keyword(s):

Cell Surface ◽

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Oxidase Activity ◽

Surface Marker ◽

Granulomatous Disease ◽

Nadph Oxidase Activity ◽

Bicistronic Vector ◽

High Level

Download Full-text

Deficiency of NADPH oxidase activity in chronic granulomatous disease

The Journal of Pediatrics ◽

10.1016/s0022-3476(77)80632-x ◽

1977 ◽

Vol 90 (2) ◽

pp. 213-217 ◽

Cited By ~ 37

Author(s):

Linda C. McPhail ◽

Lawrence R. DeChatelet ◽

Pamela S. Shirley ◽

Catherine Wilfert ◽

Richard B. Johnston ◽

...

Keyword(s):

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Oxidase Activity ◽

Granulomatous Disease ◽

Nadph Oxidase Activity

Download Full-text

Late-Onset of Chronic Granulomatous Disease Revealed By Paecilomyces Lilacinus Cutaneous Infection

10.21203/rs.3.rs-516219/v1 ◽

2021 ◽

Author(s):

Clément Lemaigre ◽

Felipe Suarez ◽

Jean-Philippe Martellosio ◽

Cindy Barbarin ◽

Kevin Brunet ◽

...

Keyword(s):

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Oxidase Activity ◽

Late Onset ◽

Asymptomatic Carrier ◽

Clinical Manifestations ◽

Female Carrier ◽

Granulomatous Disease ◽

Nadph Oxidase Activity ◽

Inherited Immunodeficiency

Abstract Chronic granulomatous disease (CGD) is an inherited immunodeficiency due to defective leukocyte NADPH responsible for recurrent infections and aberrant inflammation. Mutations in the CYBB gene are responsible for the X-linked CGD and account for approximately 70% of the cases. CGD is diagnosed during childhood in males. Female carriers may have biased X inactivation and may present with clinical manifestations depending on the level of residual NADPH oxidase activity. We report the case of a previously asymptomatic female carrier who was diagnosed at age 67 with a skin infection with the rare fungus, Paecylomyces lilacinus as the first manifestation of CGD. Dihydrorhodamin 123 (DHR) activity was below 10%. Next-generation sequencing (NGS) revealed mutations in DNMT3A, ASXL1, and STAG2 suggesting that clonal hematopoiesis of undetermined potential (CHIP) could be responsible for a progressive loss of NADPH oxidase activity and the late onset of X-linked CGD in this patient. Long-term follow-up of asymptomatic carrier women seems to be essential after 50 years old.

Download Full-text

Prolonged recombinant interferon-gamma therapy in chronic granulomatous disease: evidence against enhanced neutrophil oxidase activity

Blood ◽

10.1182/blood.v79.6.1558.bloodjournal7961558 ◽

1992 ◽

Vol 79 (6) ◽

pp. 1558-1562 ◽

Cited By ~ 1

Author(s):

RC Woodman ◽

RW Erickson ◽

J Rae ◽

HS Jaffe ◽

JT Curnutte

Keyword(s):

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Interferon Gamma ◽

Oxidase Activity ◽

Small Subset ◽

Variant Form ◽

Granulomatous Disease ◽

Recombinant Interferon ◽

Nadph Oxidase Activity ◽

O2 Production

Recombinant interferon-gamma (rIFN-gamma) therapy has become an effective form of prophylaxis for patients with chronic granulomatous disease (CGD). Preliminary studies with CGD suggested that rIFN-gamma treatment enhanced phagocyte oxidase activity and increased superoxide (O2-) production. We evaluated several aspects of neutrophil NADPH oxidase activity in 19 CGD patients (representing all four known types of CGD) receiving prolonged rIFN-gamma therapy (6 to 27 months). In contrast to earlier studies, we failed to detect any improvement in neutrophil NADPH oxidase activity in 18 of the 19 CGD patients as determined by (1) intact cell O2- production (continuous assay), (2) nitroblue tetrazolium (NBT) staining, (3) cytochrome b558 spectroscopy, and (4) activity levels of cytosol and membrane oxidase components using a cell-free activation system. One patient with a variant form of X-linked CGD had a transient increase in neutrophil O2- production following 3 months of rIFN-gamma therapy. However, this was not sustained, and was not associated with any change in cytochrome b levels. In some patients, rIFN-gamma therapy was associated with the appearance of a small subset of circulating monocytes (1% to 20%) that were NBT-positive. Although the functional significance of this monocyte subpopulation needs to be determined, these results suggest that one possible mechanism by which rIFN-gamma may benefit CGD patients is by partially correcting the respiratory burst defect in a subset of monocytes. We conclude that the clinical benefit of prolonged rIFN-gamma therapy in the vast majority of CGD patients is not due to enhanced neutrophil NADPH oxidase activity. The mechanism of action of rIFN-gamma in most CGD patients remains unknown.

Download Full-text

Retrovirus gene therapy for X-linked chronic granulomatous disease can achieve stable long-term correction of oxidase activity in peripheral blood neutrophils

Blood ◽

10.1182/blood-2009-05-222760 ◽

2010 ◽

Vol 115 (4) ◽

pp. 783-791 ◽

Cited By ~ 135

Author(s):

Elizabeth M. Kang ◽

Uimook Choi ◽

Narda Theobald ◽

Gilda Linton ◽

Debra A. Long Priel ◽

...

Keyword(s):

Gene Therapy ◽

Chronic Granulomatous Disease ◽

Oxidase Activity ◽

Severe Infection ◽

Granulomatous Disease ◽

Gene Therapy Trial ◽

Blood Neutrophils ◽

Severe Infections ◽

Therapy Trial

Abstract Chronic granulomatous disease (CGD) is associated with significant morbidity and mortality from infection. The first CGD gene therapy trial resulted in only short-term marking of 0.01% to 0.1% of neutrophils. A recent study, using busulfan conditioning and an SFFV retrovirus vector, achieved more than 20% marking in 2 patients with X-linked CGD. However, oxidase correction per marked neutrophil was less than normal and not sustained. Despite this, patients clearly benefited in that severe infections resolved. As such, we initiated a gene therapy trial for X-CGD to treat severe infections unresponsive to conventional therapy. We treated 3 adult patients using busulfan conditioning and an MFGS retroviral vector encoding gp91phox, achieving early marking of 26%, 5%, and 4% of neutrophils, respectively, with sustained long-term marking of 1.1% and 0.03% of neutrophils in 2 of the patients. Gene-marked neutrophils have sustained full correction of oxidase activity for 34 and 11 months, respectively, with full or partial resolution of infection in those 2 patients. Gene marking is polyclonal with no clonal dominance. We conclude that busulfan conditioning together with an MFGS vector is capable of achieving long-term correction of neutrophil oxidase function sufficient to provide benefit in management of severe infection. This study was registered at www.clinicaltrials.gov as #NCT00394316.

Download Full-text