title: Gene correction of induced pluripotent stem cells derived from a murine model of X-linked chronic granulomatous disorder.
abstract: Gene therapy presents an attractive alternative to allogeneic haematopoietic stem cell transplantation (HSCT) for treating patients suffering from primary immunodeficiency disorder (PID). The conceptual advantage of gene correcting a patient's autologous HSCs lies in minimizing or completely avoiding immunological complications arising from allogeneic transplantation while conferring the same benefits of immune reconstitution upon long-term engraftment. Clinical trials targeting X-linked chronic granulomatous disorder (X-CGD) have shown promising results in this context. However, long-term clinical benefits in these patients have been limited by issues of poor engraftment of gene-transduced cells coupled with transgene silencing and vector induced clonal proliferation. Novel vectors incorporating safety features such as self-inactivating (SIN) mutations in the long terminal repeats (LTRs) along with synthetic promoters driving lineage-restricted sustainable expression of the gp91phox transgene are expected to resolve the current pitfalls and require rigorous preclinical testing. In this chapter, we have outlined a protocol in which X-CGD mouse model derived induced pluripotent stem cells (iPSCs) have been utilized to develop a platform for investigating the efficacy and safety profiles of novel vectors prior to clinical evaluation.
authors.affiliation: Molecular Immunology Unit, Centre for Immunodeficiency, UCL Institute of Child Health, London, UK.
authors.forename: Sayandip
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D005260:Female; D018014:Gene Transfer Techniques; D050172:Genes, X-Linked; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006105:Granulomatous Disease, Chronic / Q000235:genetics* / Q000628:therapy*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism; D014161:Transduction, Genetic