Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome
Wiskott-Aldrich syndrome (WAS) is a severe X-linked primary immunodeficiency resulting from a diversity of mutations distributed across all 12 exons of the WAS gene. WAS encodes a hematopoietic-specific and developmentally regulated cytoplasmic protein (WASp). The objective of this study was to deve...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S232905012400024X |
| _version_ | 1797300388141989888 |
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| author | Melissa Pille John M. Avila So Hyun Park Cuong Q. Le Haipeng Xue Filomeen Haerynck Lavanya Saxena Ciaran Lee Elizabeth J. Shpall Gang Bao Bart Vandekerckhove Brian R. Davis |
| author_facet | Melissa Pille John M. Avila So Hyun Park Cuong Q. Le Haipeng Xue Filomeen Haerynck Lavanya Saxena Ciaran Lee Elizabeth J. Shpall Gang Bao Bart Vandekerckhove Brian R. Davis |
| author_sort | Melissa Pille |
| collection | DOAJ |
| description | Wiskott-Aldrich syndrome (WAS) is a severe X-linked primary immunodeficiency resulting from a diversity of mutations distributed across all 12 exons of the WAS gene. WAS encodes a hematopoietic-specific and developmentally regulated cytoplasmic protein (WASp). The objective of this study was to develop a gene correction strategy potentially applicable to most WAS patients by employing nuclease-mediated, site-specific integration of a corrective WAS gene sequence into the endogenous WAS chromosomal locus. In this study, we demonstrate the ability to target the integration of WAS2-12-containing constructs into intron 1 of the endogenous WAS gene of primary CD34+ hematopoietic stem and progenitor cells (HSPCs), as well as WASp-deficient B cell lines and WASp-deficient primary T cells. This intron 1 targeted integration (TI) approach proved to be quite efficient and restored WASp expression in treated cells. Furthermore, TI restored WASp-dependent function to WAS patient T cells. Edited CD34+ HSPCs exhibited the capacity for multipotent differentiation to various hematopoietic lineages in vitro and in transplanted immunodeficient mice. This methodology offers a potential editing approach for treatment of WAS using patient’s CD34+ cells. |
| first_indexed | 2024-03-07T23:06:27Z |
| format | Article |
| id | doaj.art-c869b7adf0164810aa0dd2b9dccebba3 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-03-07T23:06:27Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-c869b7adf0164810aa0dd2b9dccebba32024-02-22T04:52:47ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012024-03-01321101208Gene editing-based targeted integration for correction of Wiskott-Aldrich syndromeMelissa Pille0John M. Avila1So Hyun Park2Cuong Q. Le3Haipeng Xue4Filomeen Haerynck5Lavanya Saxena6Ciaran Lee7Elizabeth J. Shpall8Gang Bao9Bart Vandekerckhove10Brian R. Davis11Department of Diagnostic Sciences, Ghent University, 9000 Ghent, BelgiumBrown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, USADepartment of Bioengineering, Rice University, Houston, TX, USABrown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, USABrown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, USADepartment of Internal Medicine and Pediatrics, Ghent University, 9000 Ghent, BelgiumDepartment of Bioengineering, Rice University, Houston, TX, USADepartment of Bioengineering, Rice University, Houston, TX, USAM.D. Anderson Cancer Center, Houston, TX, USADepartment of Bioengineering, Rice University, Houston, TX, USA; Corresponding author: Gang Bao, Department of Bioengineering, Rice University, Houston, TX, USA.Department of Diagnostic Sciences, Ghent University, 9000 Ghent, Belgium; Corresponding author: Bart Vandekerckhove, Department of Diagnostic Sciences, Ghent University, 9000 Ghent, Belgium.Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, USA; Corresponding author: Brian R. Davis, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, USA.Wiskott-Aldrich syndrome (WAS) is a severe X-linked primary immunodeficiency resulting from a diversity of mutations distributed across all 12 exons of the WAS gene. WAS encodes a hematopoietic-specific and developmentally regulated cytoplasmic protein (WASp). The objective of this study was to develop a gene correction strategy potentially applicable to most WAS patients by employing nuclease-mediated, site-specific integration of a corrective WAS gene sequence into the endogenous WAS chromosomal locus. In this study, we demonstrate the ability to target the integration of WAS2-12-containing constructs into intron 1 of the endogenous WAS gene of primary CD34+ hematopoietic stem and progenitor cells (HSPCs), as well as WASp-deficient B cell lines and WASp-deficient primary T cells. This intron 1 targeted integration (TI) approach proved to be quite efficient and restored WASp expression in treated cells. Furthermore, TI restored WASp-dependent function to WAS patient T cells. Edited CD34+ HSPCs exhibited the capacity for multipotent differentiation to various hematopoietic lineages in vitro and in transplanted immunodeficient mice. This methodology offers a potential editing approach for treatment of WAS using patient’s CD34+ cells.http://www.sciencedirect.com/science/article/pii/S232905012400024Xgene editinggene therapyprimary immune diseaseWiskott-Aldrich syndromeimmunology |
| spellingShingle | Melissa Pille John M. Avila So Hyun Park Cuong Q. Le Haipeng Xue Filomeen Haerynck Lavanya Saxena Ciaran Lee Elizabeth J. Shpall Gang Bao Bart Vandekerckhove Brian R. Davis Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome Molecular Therapy: Methods & Clinical Development gene editing gene therapy primary immune disease Wiskott-Aldrich syndrome immunology |
| title | Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome |
| title_full | Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome |
| title_fullStr | Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome |
| title_full_unstemmed | Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome |
| title_short | Gene editing-based targeted integration for correction of Wiskott-Aldrich syndrome |
| title_sort | gene editing based targeted integration for correction of wiskott aldrich syndrome |
| topic | gene editing gene therapy primary immune disease Wiskott-Aldrich syndrome immunology |
| url | http://www.sciencedirect.com/science/article/pii/S232905012400024X |
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