Suchergebnisse

This work was supported by the Spanish ISCIII Health Research Fund and the European Regional Development Fund (FEDER) through research grants PI12/01097, PI15/02015, and PI18/00337 (to F.M.). The CECEyU and CSyF of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia provided the following research grants: 2016000073391-TRA, 2016000073332-TRA, PI-57069, and PAIDI-Bio326 (to F.M.) and PI-0407/2012 (to P.M). P.M. was supported by the European Union's Horizon 2020 through grant agreement no. 329284 (People Marie Curie Actions, Intra-European Fellowship [IEF], call: FP7-PEOPLE-2012-IEF) and by the Spanish ISCIII Health Research Fund through a postdoctoral fellowship "Sara Borrell" (CD09/00200). M.T.M. is funded by MCI through a fellowship FPU16/05467. A.G. is supported by funds from AFM-Telethon. We thank Dr. Claire Booth and Dr. Inmaculada Herrera for providing HDs and WAS patient blood samples and Ailsa Greppy for technical support with the animals (Western Labs, ICH-UCL). Some figures were created with BioRender.com. ; Supplemental Information can be found online at https://doi.org/10. 1016/j.omtm.2020.09.006. ; 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. ; Spanish 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

Psoriasis is a chronic inflammatory skin disease that speeds up the life cycle of skin cells, forming scales and red patches that are itchy and sometimes painful. It is a complex disease of autoimmune origin and genetic predisposition with more than 10 different loci associated. Here we described the production of an iPSC line generated by Sendai Virus (Klf4, Oct3/4, Sox2 and c-Myc) reprogramming of Peripheral Blood Mononuclear Cells (PBMCs) from a Psoriasis patient. The iPSC line generated has normal 46XY karyotype, is free of SeV genome and transgenes insertions, express high levels of pluripotency markers and can differentiate into all three germ layers. ; Funding was provided by the Spanish Ministry for Science and Innovation (RTC-2016-5324-1). JMMF was a postdoctoral Berrikertu fellow from the Basque government. AF was a recipient of Juan de la Cierva (JCI-2006-2675) and Torres Quevedo (PTQ-16-08496) postdoctoral fellowships from the Spanish Ministry for Science and Innovation.

Cystic fibrosis (CF) is the main genetic cause of death among the Caucasian population. The disease is characterized by abnormal fluid and electrolyte mobility across secretory epithelia. The first manifestations occur within hours of birth (meconium ileus), later extending to other organs, generally affecting the respiratory tract. It is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR encodes a cyclic adenosine monophosphate (cAMP)-dependent, phosphorylation-regulated chloride channel required for transport of chloride and other ions through cell membranes. There are more than 2,000 mutations described in the CFTR gene, but one of them, phenylalanine residue at amino acid position 508 (p.F508del), a recessive allele, is responsible for the vast majority of CF cases worldwide. Here, we present the results of the application of genome-editing techniques to the restoration of CFTR activity in p.F508del patient-derived induced pluripotent stem cells (iPSCs). Gene-edited iPSCs were subsequently used to produce intestinal organoids on which the physiological activity of the restored gene was tested in forskolin-induced swelling tests. The seamless restoration of the p.F508del mutation resulted in normal expression of the mature CFTR glycoprotein, full recovery of CFTR activity, and a normal response of the repaired organoids to treatment with two approved CF therapies: VX-770 and VX-809. ; This work was supported by grants from the Spanish Ministry for Science and Innovation (PLE2009-0091, RTC-2014-2207-1, and IPT-2011-1402-900000); ISCIII PI14/01073, cofunded by ERDF/ESF "Investing in your future"; the Balearic Government (16023/2008); the Spanish Cystic Fibrosis Federation (Pablo Motos Grant); Federación ASEM-Telemaratón "Todos somos raros, todos somos únicos"; Fundación Salud 2000; the European Commission (H2020, PHC-667079); and an endowment from METROVACESA. J.M.M.-F. was a postdoctoral Berrikertu fellow granted by the Basque Government. A.F. was a recipient of Juan de la Cierva (JCI-2006-2675) and Torres Quevedo (PTQ-16-08496) postdoctoral fellowships from the Spanish Ministry for Science and Innovation.

The mutation E280A in PSEN1 (presenilin-1) is the most common cause of early-onset familial Alzheimer's Disease (fAD). It presents autosomal dominant inheritance and frequently leads to the manifestation of the disease in relatively young individuals. Here we report the generation of one PSEN1 E280A iPSC line derived from an early-onset patient. OriP/EBNA1-based episomal plasmids containing OCT3/4, SOX2, KLF4, L-MYC, LIN28, BCL-xL and shp53 were used to reprogram oral mucosa fibroblasts. The iPSC line generated has normal karyotype, carry the E280A mutation, is free of plasmid integration, express high levels of pluripotency markers and can differentiate into all three germ layers. ; Funding was provided by the Basque Government grant: Elkartek 20017 (DRUG4AD).