WAS Promoter-Driven Lentiviral Vectors Mimic Closely the Lopsided WASP Expression during Megakaryocytic Differentiation
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AuthorMuñoz Fernández, Pilar; Tristán Manzano, María; Sánchez Gilabert, Almudena; Santilli, Giorgia; Galy, Anne; Thrasher, Adrian J.; Martín Molina, Francisco
Muñoz, P., Tristán-Manzano, M., Sánchez-Gilabert, A., Santilli, G., Galy, A., Thrasher, A. J., & Martin, F. (2020). WAS Promoter-Driven Lentiviral Vectors Mimic Closely the Lopsided WASP Expression during Megakaryocytic Differentiation. Molecular Therapy-Methods & Clinical Development, 19, 220-235. [https://doi.org/10.1016/j.omtm.2020.09.006]
SponsorshipSpanish ISCIII Health Research Fund; European Commission PI12/01097 PI15/02015 PI18/00337; CSyF of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia 2016000073391-TRA 2016000073332-TRA PI-57069 PAIDI-Bio326 PI-0407/2012; European Commission 329284; MCI FPU16/05467; Association Francaise contre les Myopathies; Spanish ISCIII Health Research Fund through a postdoctoral fellowship "Sara Borrell" CD09/00200; CECEyU of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia 2016000073391-TRA 2016000073332-TRA PI-57069 PAIDI-Bio326 PI-0407/2012
Transplant of gene-modified autologous hematopoietic progenitors cells has emerged as a new therapeutic approach for Wiskott-Aldrich syndrome (WAS), a primary immunodeficiency with microthrombocytopenia and abnormal lymphoid and myeloid functions. Despite the clinical benefits obtained in ongoing clinical trials, platelet restoration is suboptimal. The incomplete restoration of platelets in these patients can be explained either by a low number of corrected cells or by insufficient or inadequate WASP expression during megakaryocyte differentiation and/or in platelets. We therefore used in vitro models to study the endogenous WASP expression pattern during megakaryocytic differentiation and compared it with the expression profiles achieved by different therapeutic lentiviral vectors (LVs) driving WAS cDNA through different regions of the WAS promoter. Our data showed that all WAS promoter-driven LVs mimic very closely the endogenous WAS expression kinetic during megakaryocytic differentiation. However, LVs harboring the full-length (1.6-kb) WAS-proximal promoter (WW1.6) or a combination of the WAS alternative and proximal promoters (named AW) had the best behavior. Finally, all WAS-driven LVs restored the WAS knockout (WASKO) mice phenotype and functional defects of hematopoietic stem and progenitor cells (HSPCs) from a WAS patient with similar efficiency. In summary, our data back up the use of WW1.6 and AW LVs as physiological gene transfer tools for WAS therapy.