Project description:Huntington's disease (HD) and control GLAST-postive induced pluripotent stem cell (iPSC)-derived astrocytes underwent single-nucleus RNA-sequencing to investigate cell state diversity across control and HD patient-derived astrocytes.
Project description:Huntington's disease is caused by an expanded CAG repeat in the huntingtin gene, yeilding a Huntingtin protein with an expanded polyglutamine tract. Patient-derived induced pluripotent stem cells (iPSCs) can help understand disease; however, defining pathological biomarkers in challanging. Here we used LC-MS/MS to determine differences in mitochondrial proteome between iPSC-derived neurons from healthy donors and Huntington's disease patients.
Project description:Compared the global gene expression profiles of HD- and CON-iPSC-derived neurons We used microarrays to detail the global programme of gene expression for comparing the global gene expression profiles of HD- and CON-iPSC-derived neurons and facilitating studies of medium spiny neurons (MSN)-degenerative processes of Huntington's Disease (HD). By using a step-wise in vitro differentiation protocol combining EB formation, neural induction by small molecules, treatment with inhibitors of the TGFß pathway (SB431542) and the BMP pathway (LDN193189), and mechanical isolation/purification of neural progenitors and neurons, we induced 60-70% of control iPSCs or HD-iPSCs to differentiate into GABA- and DARPP-32- double positive neurons.
Project description:To investigate the transcriptomic profile and molecular machnisiums using RNA sequecning analysis for the iPSC derived neural cells and brain organoids from the patient of Alpers’ Disease We then performed gene expression profiling analysis using data obtained from RNA-seq of the iPSC derived neural cells and brain organoids from the patient of Alpers’ Disease and healthy controls
Project description:Huntington's Disease (HD) is caused by a CAG expansion in the huntingtin gene. Expansion of the polyglutamine tract in the huntingtin protein results in massive cell death in the striatum of HD patients. We report that human induced pluripotent stem cells (iPSCs) derived from HD patient fibroblasts can be corrected by replacing the expanded CAG repeat with a normal repeat using homologous recombination, and that the correction persists in iPSC differentiation into DARPP-32 positive neurons in vitro and vivo. Further, correction of the HD-iPSCs normalized pathogenic HD signaling pathways (cadherin, TGF-?, BNDF, caspase activation), and reversed disease phenotypes such as susceptibility to cell death and altered mitochondrial bioenergetics in neural stem cells. The ability to make patient-specific, genetically corrected iPSCs from HD patients will provide relevant disease models in identical genetic backgrounds and is a critical step for the eventual use of these cells in cell replacement therapy. 16 experimental samples were used overall. There were 8 replicates per group, with one group being the control, and the other being the experimental. Comparison was carried out on the Nimblegen platform.
