Lentiviral gene therapy for X-linked chronic granulomatous disease recapitulates endogenous CYBB regulation and expression

•We used a bioinformatics-guided approach to design a lentiviral vector driven by endogenous enhancer and promoter elements of the CYBB gene.•Our novel vector restores physiologic regulation and expression of CYBB for the treatment of X-CGD. [Display omitted] X-linked chronic granulomatous disease (...

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Published in:	Blood Vol. 141; no. 9; pp. 1007 - 1022
Main Authors:	Wong, Ryan L., Sackey, Sarah, Brown, Devin, Senadheera, Shantha, Masiuk, Katelyn, Quintos, Jason P., Colindres, Nicole, Riggan, Luke, Morgan, Richard A., Malech, Harry L., Hollis, Roger P., Kohn, Donald B.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 02-03-2023
Subjects:	Animals Genetic Therapy - methods Granulomatous Disease, Chronic - genetics Granulomatous Disease, Chronic - therapy Mice Mutation NADPH Oxidase 2 - genetics NADPH Oxidases - genetics NADPH Oxidases - metabolism
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Summary:	•We used a bioinformatics-guided approach to design a lentiviral vector driven by endogenous enhancer and promoter elements of the CYBB gene.•Our novel vector restores physiologic regulation and expression of CYBB for the treatment of X-CGD. [Display omitted] X-linked chronic granulomatous disease (X-CGD) is a primary immunodeficiency caused by mutations in the CYBB gene, resulting in the inability of phagocytic cells to eliminate infections. To design a lentiviral vector (LV) capable of recapitulating the endogenous regulation and expression of CYBB, a bioinformatics-guided approach was used to elucidate the cognate enhancer elements regulating the native CYBB gene. Using this approach, we analyzed a 600-kilobase topologically associated domain of the CYBB gene and identified endogenous enhancer elements to supplement the CYBB promoter to develop MyeloVec, a physiologically regulated LV for the treatment of X-CGD. When compared with an LV currently in clinical trials for X-CGD, MyeloVec showed improved expression, superior gene transfer to hematopoietic stem and progenitor cells (HSPCs), corrected an X-CGD mouse model leading to complete protection against Burkholderia cepacia infection, and restored healthy donor levels of antimicrobial oxidase activity in neutrophils derived from HSPCs from patients with X-CGD. Our findings validate the bioinformatics-guided design approach and have yielded a novel LV with clinical promise for the treatment of X-CGD. X-linked chronic granulomatous disease is caused by mutations in the CYBB gene, encoding a subunit of the nicotinamide adenine dinucleotide phosphate oxidase complex. It is a target for gene therapy; however, current lentiviral vectors have limited success in reconstituting oxidase activity. Wong et al used bioinformatics to define enhancer elements and create a novel lentiviral vector that mimics physiologic regulation of CYBB. In preclinical studies, the vector showed improved gene transfer and reconstitution of oxidase activity.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0006-4971 1528-0020 1528-0020
DOI:	10.1182/blood.2022016074