Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model

Toscano, Miguel G.; Muñoz, Pilar; Sánchez‐Gilabert, Almudena; Cobo, Manuel J.; Benabdellah, Karim; Anderson, Per; Ramos-Mejía, Verónica; Real, Pedro J.; Neth, Olaf; Molinos-Quintana, Águeda; Gregory, Philip D.; Holmes, Michael C.; Martín, Francisco

doi:10.1038/mt.2015.196

Cited by 8 publications

(10 citation statements)

References 45 publications

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“…Dysfunctional MKs and PLTs have also been observed from normal embryonic stem cell lines in which the WAS locus was targeted by zinc finger nucleases. Gene‐edited MKs and especially PLTs displayed altered phenotypes and defective responses to agonists . Conversely, normal maturation and migration to stromal‐derived factor‐1α of MKs differentiated from CD34 + cells that were isolated from patients with WAS and normal pro‐PLT formation have also been reported in the literature .…”

Section: Wasp and Pltsmentioning

confidence: 98%

“…Geneedited MKs and especially PLTs displayed altered phenotypes and defective responses to agonists. 105 Conversely, normal maturation and migration to stromal-derived factor-1 of MKs differentiated from CD34 + cells that were isolated from patients with WAS and normal pro-PLT formation have also been reported in the literature. 106 In line with these results, pro-PLT formation in MKs has recently been demonstrated to be N-WASp and not WASp dependent.…”

Section: Pathogenesis Of Thrombocytopenia In Wasmentioning

confidence: 99%

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Platelets in Wiskott-Aldrich syndrome: Victims or executioners?

Sereni

Castiello

Villa

2017

Journal of Leukocyte Biology

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Microthrombocytopenia is the clinical hallmark of WAS, a rare X-linked immunodeficiency that is characterized by eczema, autoimmunity, and cancer susceptibility. This disease is caused by mutations in the WAS gene, which is expressed in hematopoietic cells and regulates actin cytoskeleton remodeling thereby modulating various cellular functions, including motility, immunologic synapse assembly, and signaling. Despite extensive studies that have provided great insight into the relevance of this molecule to innate and cellular immunity, the exact mechanisms of microthrombocytopenia in WAS are still unknown. This review focuses on the recent progress made in dissecting the pathogenesis of platelet defects in patients with WAS and their murine counterparts. In parallel, we will provide an overview of the state-of-the art platelets as immune modulators at the interface between hemostasis and the immune system, which suggests that these cells may have a direct role in the pathogenesis of immune dysregulation in WAS.

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Section: Wasp and Pltsmentioning

confidence: 98%

Section: Pathogenesis Of Thrombocytopenia In Wasmentioning

confidence: 99%

Platelets in Wiskott-Aldrich syndrome: Victims or executioners?

Sereni

Castiello

Villa

2017

Journal of Leukocyte Biology

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“…Taking these data into consideration, the authors managed to demonstrate that mutation in LRRK2 induced dysregulation of CPNE8, MAP7, UHRF2, ANXA1 and CADPS2 genes. Other good examples of the relevance of using isogenic cell lines were showed by Li et al [61] and by our group [62]. Li et al generated a model for the Rett Syndrome by mutating the MECP2 gene.…”

Section: Editing Hescs For Disease Modelingmentioning

confidence: 97%

“…MECP2 mutant neurons mimic the defects observed in this disorder and unbiased global gene expression analyses (thanks to the use of isogenic cell lines) showed that MECP2 protein functions as a global gene activator in neurons but not in neural precursors. Similarly, our group has generated human cellular models for Wiskott-Aldrich syndrome (WAS) by mutating the WAS gene in hESCs using ZFNs [62]. Using these models, we uncovered that the absence of WAS protein also affected early hematopoiesis and megakaryocyte development, a phenotype that could not be observed when using iPSCs from WAS patients [63].…”

Section: Editing Hescs For Disease Modelingmentioning

confidence: 99%

Stem Cells for Modeling Human Disease

Martín¹,

Sánchez‐Gilabert²,

Tristán‐Manzano³

et al. 2016

Pluripotent Stem Cells - From the Bench to the Clinic

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Human pluripotent stem cells (PSCs) in the form of human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) are capable of growing indefinitely in vitro, maintaining their capacity to differentiate into the three primary germ layers: mesoderm, endoderm and ectoderm. Different protocols have been developed to differentiate PSCs into almost any cellular type with different degree of success. This technology has allowed scientists to use patient-derived iPSCs to study the physiopathology of the disease by analyzing the phenotype of the cells derived from these iPSCs. However, control iPSCs obtained from healthy individuals will always have different genomic environment than patient's iPSCs, making it difficult the interpretation of the cells phenotype. The recent appearance of specific nucleases [zinc-finger nucleases (ZFNs), the transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)] has made it possible to edit the genome of PSCs. We can now generate syngeneic hESCs or iPSCs harboring the desired mutation and comparing the emerging cells with those derived from genetically identical PSCs that will differ only in the mutated gene. In this chapter, we summarize the progress made in this field and discuss the different approaches that have been used recently for the generation of syngeneic human pluripotent cellular models for different pathologies.

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“…The exact functions of WASP in PLTs remains largely unknown, but there is strong evidence suggesting that it plays a critical role regulating MK differentiation and PLT formation by inhibiting these processes in the absence of the appropriated signals. 18 , 19 , 20 This could sound contradictory with the thrombocytopenia found in WAS patients, but it is actually a potential explanation for it. Indeed, PLTs produced in the absence of WASP have not been developed properly and have a reduced size, abnormal ultrastructure, and surface markers that lead to their elimination in the spleen and other tissues.…”

Section: Introductionmentioning

confidence: 93%

WAS Promoter-Driven Lentiviral Vectors Mimic Closely the Lopsided WASP Expression during Megakaryocytic Differentiation

Muñoz

Tristán‐Manzano

Sánchez‐Gilabert

et al. 2020

Molecular Therapy - Methods & Clinical Development

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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.

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Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model

Cited by 8 publications

References 45 publications

Platelets in Wiskott-Aldrich syndrome: Victims or executioners?

Platelets in Wiskott-Aldrich syndrome: Victims or executioners?

Stem Cells for Modeling Human Disease

WAS Promoter-Driven Lentiviral Vectors Mimic Closely the Lopsided WASP Expression during Megakaryocytic Differentiation

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