Gene Correction of Wiskott–Aldrich syndrome iPS Cells Rescues Proplatelet Defects and Improves Platelet Size

Praewphan Ingrungruanglert; Sarinya Phodang; Pramuk Amarinthnukrowh; Phattarawan Meehart; Pornpitra Pratedrat; Narissara Suratannon; Vorasuk Shotelersuk; Kanya Suphapeetiporn; Nipan Israsena

doi:10.1055/a-2508-0983

Thrombosis and Haemostasis, Inhaltsverzeichnis

CC BY-NC-ND 4.0 · Thromb Haemost
DOI: 10.1055/a-2508-0983

Cellular Haemostasis and Platelets

Gene Correction of Wiskott–Aldrich syndrome iPS Cells Rescues Proplatelet Defects and Improves Platelet Size

Praewphan Ingrungruanglert

¹Center of Excellence for Stem Cell and Cell Therapy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

²Excellence Center for Stem Cell and Cell Therapy, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Sarinya Phodang

¹Center of Excellence for Stem Cell and Cell Therapy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

²Excellence Center for Stem Cell and Cell Therapy, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Pramuk Amarinthnukrowh

³Department of Pediatrics, Faculty of Medicine, Center of Excellence for Medical Genomics, Medical Genomics Cluster, Chulalongkorn University, Bangkok, Thailand

⁴Excellence Center for Genomics and Precision Medicine, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Phattarawan Meehart

¹Center of Excellence for Stem Cell and Cell Therapy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

²Excellence Center for Stem Cell and Cell Therapy, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Pornpitra Pratedrat

⁵Department of Basic Medical Science, Faculty of Medicine Vajira Hospital, Navamindradhiraj University, Bangkok, Thailand

,

Narissara Suratannon

⁶Division of Allergy, Immunology, and Rheumatology, Department of Pediatrics, Faculty of Medicine, Center of Excellence for Allergy and Clinical Immunology, Chulalongkorn University, Bangkok, Thailand

⁷King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Vorasuk Shotelersuk

³Department of Pediatrics, Faculty of Medicine, Center of Excellence for Medical Genomics, Medical Genomics Cluster, Chulalongkorn University, Bangkok, Thailand

⁴Excellence Center for Genomics and Precision Medicine, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Kanya Suphapeetiporn

³Department of Pediatrics, Faculty of Medicine, Center of Excellence for Medical Genomics, Medical Genomics Cluster, Chulalongkorn University, Bangkok, Thailand

⁴Excellence Center for Genomics and Precision Medicine, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

,

Nipan Israsena

¹Center of Excellence for Stem Cell and Cell Therapy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

²Excellence Center for Stem Cell and Cell Therapy, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand

⁸Department of Pharmacology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

› Institutsangaben

Abstract

Wiskott–Aldrich syndrome (WAS) is a severe X-linked disorder caused by loss-of-function mutations in the WAS gene, responsible for encoding WAS protein (WASP), a key regulator of the actin cytoskeleton in all hematopoietic cells, except red blood cells. The mechanism underlying microthrombocytopenia, a distinctive feature of WAS and a major contributor to mortality, remains not fully elucidated. In this study, using different gene-editing strategies, we corrected mutations in patient-derived WAS-induced pluripotent stem cell (iPSC) lines, generating isogeneic WAS-iPSC lines. These included lines with direct mutation-specific correction and lines incorporating a WASP transgene cassette regulated by the MND or WAS1.6 kb promoter integrated at the safe harbor AAV1 site. Our results demonstrated that direct mutation correction successfully restored WASP levels to the equivalent of the wild-type in iPSC-derived megakaryocytes (MKs). In contrast, the AAV1-targeted strategy using the MND and WAS1.6 promoters yielded a lower level of WASP. Notably, only the mutation-specific correction lines exhibited improvements in proplatelet structures and generated larger-sized platelets. Our findings underscore the crucial roles of WASP during human thrombopoiesis and suggest that therapeutic approaches, such as direct gene correction, which can achieve physiologic levels of WASP in MKs, hold promise for ameliorating platelet defects in individuals with WAS.

Keywords

thrombocytopenia - Wiskott–Aldrich syndrome - gene targeting

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Referenzen

References
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