GSE105041
hESC-based human glial chimeric mice reveal glial differentiation defects in Huntington disease
Huntington's disease (HD) is characterized by hypomyelination as well as by neuronal loss. To assess the basis for white matter involution in HD, we generated bipotential glial progenitor cells (GPCs) from human embryonic stem cells (hESCs), derived from either huntingtin (mHTT)-mutant embryos or normal controls, and performed RNA sequence analysis to assess mHTT-dependent changes in gene expression. In hGPCs derived from 3 distinct mHTT-expressing hESC lines, a set of transcription factors associated with glial differentiation and myelin synthesis was sharply down-regulated, relative to normal hESC GPCs. In particular, NKX2.2, OLIG2, SOX10 and MYRF were all suppressed, with the consequent diminution of myelinogenesis-associated transcription. Accordingly, when mHTT-expressing hGPCs were transplanted into hypomyelinated shiverer mice, the resultant mHTT glial chimeras were hypomyelinated. The mHTT hGPCs also manifested impaired astrocytic differentiation, and developed abnormal fiber domain architecture. These data suggest that white matter involution in HD is a product of a cell-autonomous mHTT-dependent suppression of both astrocytic and oligodendrocytic differentiation by affected GPCs. Overall design: Embryonic stem cells (hESCs) derived from 3 Huntington's disease embryos (designated to HD lines 17, 18, and 20) and 2 healthy control embryos (designated to CTR lines 02 and 19) were obtained from Genea Biocells, Sydney, Australia (http://geneabiocells.com/services/shelf-products/human-embryonic-stem-cells/). The hESC lines were differentiated into glia by previously described methods (Wang et al., 2013) and further purified by FACS targeting CD140a for enriched populations of Glial Progenitor Cells (GPCs) and CD44 for enriched populations of Astrocyte Progenitor Cells (APCs). The purified glial cell populations where then used in mRNA isolation by PolyA selection and mRNA sequencing analysis. Please note that the FASTQ files in the submission were edited by Trimmomatic. The software is used to trim adapter sequences and low-quality bases (see processing steps). The overall file structure and format remained unchanged. The count matrix in the submission is as obtained directly from featureCounts tool, that is, no manipulation was performed to the count data. The data normalization was performed internally in the analysis but the raw count data were used in differential expression analysis. Please refer to the Materials and Methods section in supplementary files. Additionally, the complete workflow, including R scripts and count matrix, was deposited to https://github.com/cbtncph/HD-Glial-Differentiation-Block-Goldman-Lab-2017.