1. Generation of Human Induced Pluripotent Stem Cell‐Derived Bona Fide Neural Stem Cells for Ex Vivo Gene Therapy of Metachromatic Leukodystrophy
- Author
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Francesco Morena, Serena Giannelli, Vania Broccoli, Giacomo Frati, Marco Luciani, Angela Gritti, Chiara Cavazzin, Marianna Paulis, Davide Sala, Francesca Sanvito, Alessandro Bulfone, Silvia De Cicco, Wieslawa Mentzen, Vasco Meneghini, Sabata Martino, and Anna Villa
- Subjects
0301 basic medicine ,Genetic enhancement ,Intracerebral transplantation ,Mice, SCID ,Translational Research Articles and Reviews ,Neural Stem Cells ,Cell Movement ,Mice, Inbred NOD ,Cellular Reprogramming Techniques ,Induced pluripotent stem cell ,Oligodendrocytes ,Gene Expression Regulation, Developmental ,Cell Differentiation ,General Medicine ,Cellular Reprogramming ,Neural stem cell ,3. Good health ,Phenotype ,Enzyme Induction ,Stem cell ,Reprogramming ,Pluripotent Stem Cells ,metabolic restoration ,Induced Pluripotent Stem Cells ,Biology ,Metachromatic leukodystrophy ,Cell Line ,03 medical and health sciences ,Gene therapy ,medicine ,Animals ,Humans ,Progenitor cell ,Arylsulfatase A activity ,Cerebroside-Sulfatase ,Sulfoglycosphingolipids ,Cell Biology ,Genetic Therapy ,Leukodystrophy, Metachromatic ,medicine.disease ,Coculture Techniques ,Neural stem cells ,Pluripotent stem cells ,Nerve Regeneration ,Transplantation ,Disease Models, Animal ,030104 developmental biology ,Immunology ,Cancer research ,Transcriptome ,Developmental Biology ,Stem Cell Transplantation - Abstract
Allogeneic fetal-derived human neural stem cells (hfNSCs) that are under clinical evaluation for several neurodegenerative diseases display a favorable safety profile, but require immunosuppression upon transplantation in patients. Neural progenitors derived from patient-specific induced pluripotent stem cells (iPSCs) may be relevant for autologous ex vivo gene-therapy applications to treat genetic diseases with unmet medical need. In this scenario, obtaining iPSC-derived neural stem cells (NSCs) showing a reliable “NSC signature” is mandatory. Here, we generated human iPSC (hiPSC) clones via reprogramming of skin fibroblasts derived from normal donors and patients affected by metachromatic leukodystrophy (MLD), a fatal neurodegenerative lysosomal storage disease caused by genetic defects of the arylsulfatase A (ARSA) enzyme. We differentiated hiPSCs into NSCs (hiPS-NSCs) sharing molecular, phenotypic, and functional identity with hfNSCs, which we used as a “gold standard” in a side-by-side comparison when validating the phenotype of hiPS-NSCs and predicting their performance after intracerebral transplantation. Using lentiviral vectors, we efficiently transduced MLD hiPSCs, achieving supraphysiological ARSA activity that further increased upon neural differentiation. Intracerebral transplantation of hiPS-NSCs into neonatal and adult immunodeficient MLD mice stably restored ARSA activity in the whole central nervous system. Importantly, we observed a significant decrease of sulfatide storage when ARSA-overexpressing cells were used, with a clear advantage in those mice receiving neonatal as compared with adult intervention. Thus, we generated a renewable source of ARSA-overexpressing iPSC-derived bona fide hNSCs with improved features compared with clinically approved hfNSCs. Patient-specific ARSA-overexpressing hiPS-NSCs may be used in autologous ex vivo gene therapy protocols to provide long-lasting enzymatic supply in MLD-affected brains.
- Published
- 2016