Ex Vivo Gene Therapy Using Patient iPSC-Derived NSCs Reverses Pathology in the Brain of a Homologous Mouse Model

Abstract

SummaryNeural stem cell (NSC) transplantation is a promising strategy for delivering therapeutic proteins in the brain. We evaluated a complete process of ex vivo gene therapy using human induced pluripotent stem cell (iPSC)-derived NSC transplants in a well-characterized mouse model of a human lysosomal storage disease, Sly disease. Human Sly disease fibroblasts were reprogrammed into iPSCs, differentiated into a stable and expandable population of NSCs, genetically corrected with a transposon vector, and assessed for engraftment in NOD/SCID mice. Following neonatal intraventricular transplantation, the NSCs engraft along the rostrocaudal axis of the CNS primarily within white matter tracts and survive for at least 4 months. Genetically corrected iPSC-NSCs transplanted post-symptomatically into the striatum of adult Sly disease mice reversed neuropathology in a zone surrounding the grafts, while control mock-corrected grafts did not. The results demonstrate the potential for ex vivo gene therapy in the brain using human NSCs from autologous, non-neural tissues.

Integrated analysis of proteome and transcriptome changes in the mucopolysaccharidosis type VII mouse hippocampus

“…The hippocampus was selected because it has been studied in this model both for histopathology [3, 9, 10, 24] and behavioral abnormalities [4-6, 25]. The B6 strain of MPS VII mouse was chosen due to the severity of disease in order to maximize the differences between normal and MPS VII hippocampi [24].…”

“…We focused the analysis on a single region because transcriptomic analysis in this model showed not only numerous changes between normal and diseased brains, but also significant differences in the changes between brain regions [8]. We chose the hippocampus because of the extensive histopathology associated with this region [3, 9, 10]. …”

“…MPS VII mice and normal controls were generated from the carrier strain B6.C-H2-Kbm1 /ByBir-Gusb mps/+/J [9] and were maintained in our breeding colony through carrier-carrier brother-sister mating. Identification of the MPS VII-affected mice was verified by PCR genotyping, as described previously [10].…”

“…To further assess the changes associated with the MPS VII brain, the present study directly compared proteomics and transcriptomic analyses in the hippocampus from the MPS VII mouse on the C57Bl/6 (B6) strain background in which neurodegeneration has been studied [3, 9, 10]. The combined results of the proteome and transcriptome changes were in functional categories consistent with many of the known histopathology findings [3, 9, 10]. The analyses extended findings of pathological alterations in some non-overlapping areas and provided information on the molecular manifestation of MPS VII disease between strains of mice.…”

Gene-modified, cell-based therapies—an overview

“…Transposon systems like Sleeping Beauty combine the advantages of viral and nonviral platforms: they allow expression of the transgene and transposon system using plasmids delivered as naked DNA, while also allowing sustained expression of the construct following integration into the genome [55]. T cells [56], induced pluripotent cells [57], and muscle cells [58] for potential use as cell therapies have all been successfully modified using transposon systems. Alternatively, the addition of S/MAR (scaffold/matrix associated regions) in the construct allows attachment of episomal DNA into the nuclear matrix.…”