Impact of KIF26A loss in human forebrain organoids
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ABSTRACT: KIF26A is an unconventional kinesin that lacks motor activity. To characterize its role in brain development, we generated KIF26A knock-out iPSC lines and differentiated forebrain organoids. We performed single cell RNA sequencing at Day 60 and Day 80 to compare between control and KIF26A-deleted organoids at the molecular level, which revealed cell type-specific transcriptional changes in MAPK, MYC and E2F pathways.
Project description:We performed single-cell RNA sequencing of dorsal forebrain organoids at day 53 of differentiation upon treatment with Hyper-IL-6. The study aimed at investigation of the effects of Hyper-IL-6 on transcriptional profiles of dorsal forebrain organoids at single-cell level.
Project description:Description: RNA-seq of total RNA isolated from dorsal forebrain organoids at days 50 and 55 of differentiation. The study aimed at investigating the effects of 5 or 10 days of Hyper-IL-6 exposure on transcriptional profiles of dorsal forebrain organoids.
Project description:Single cell RNA-sequencing of 90-day old astrocyte conditioned medium treated forebrain organoids (ACMOs) and control forebrain organoids
Project description:The scRNA-seq analysis of 90-day old ACMOs and forebrain organoids reveals six well-defined major clusters. All cells were categorized into six distinct clusters: (1) deep layer subcortical projection neurons, (2) upper layer projection neurons, (3) intermediate progenitors, (4) radial glial cells, (5) dividing cells, (6) astrocytes. Both ACMOs and control organoids underwent a correct trajectory of cell diversification and contained a large diversity of progenitor and neuronal cell types belonging to the neuroectodermal lineage. Transcriptomic comparisons and pathway enrichment analysis were performed between control organoids and ACMOs.
Project description:We established PCCB knockdown human induced pluripotent stem cell (hiPSC) line using CRISPR interference (CRISPRi). The established PCCB knockdown and control hiPSCs were then used to generate human forebrain organoids (hFOs). On day 60 of organoid culture, PCCB knockdown and control hFOs were randomly selected for RNA-sequencing (RNA-seq). We found that differentially expressed genes (DEGs) affected by PCCB knockdown were enriched with GABAergic synapse, synaptic vesicle cycle, neurotransmitter transport, forebrain development, axon development, synaptic organization, and calcium signaling pathways. The DEGs were also significantly overlapped with schizophrenia (SCZ)-associated genes, including genes dysregulated in brains or organoids derived from SCZ patients, and genes reported in SCZ GWAS.
Project description:Temporal dynamics of prenatal brain development are influenced by changes in the microenvironment. Particularly, extracellular proteins contribute to the dynamic niche that balances neural stem cell proliferation and differentiation. Here, we present a resource for proteome and secretome analysis of human induced pluripotent stem cell-derived dorsal forebrain organoids over the early developmental period. We used liquid chromatography-mass spectrometry to identify proteins found in whole organoid and secreted proteins at days 20, 35, and 50 of dorsal forebrain organoid differentiation. We show that the whole organoid proteome demonstrates progression in the neurodevelopmental trajectory with reduced proliferation and increased neural differentiation over time. However, secretome analysis revealed a unique signature for developmental progression. Cell adhesion molecules are enriched in the secretome of day 35 organoids, while secretome of day 50 organoids is enriched with extracellular matrix proteins, demonstrating a change in the extracellular interactions over time of organoid differentiation and maturation. We, therefore, show the relevance of secretome analysis for the thorough study of extracellular matrix-related proteins and the importance of time course study of neural organoids to understand the subtle changes that guide human neurodevelopment.
Project description:Human forebrain organoids (day 42) derived from U1M and U2F iPS cell lines were exposed with or without 1 mM valproic acid (VPA) for 72h. Organoids were then randomly selected for RNA sequencing, we found that differentially expressed genes (DEGs) induced by VPA exposure that shared in both U1M and U2F were enriched with neural development, synaptic transmission, calcium and potassium signaling pathways. The DEGs were also significantly enriched with autism spectrum disorder (ASD)-associated genes, including genes dysregulated in brains or organoids derived from ASD patients, and known ASD risk genes, as well as genes in ASD risk-associated gene coexpression modules.
Project description:Human forebrain organoids (day 50) derived from U1M and U2F iPS cell lines were exposed with or without 1 mM valproic acid (VPA) for 72h. Organoids were then used for single cell RNA sequencing (scRNA-seq). Through data processing, ten major cell types including astrocyte, choroid plexus, endothelia, intermediate progenitor cells, medial ganglionic eminence, radial glia, immature neuron, excitatory and inhibitory neuron, and microglia-like cells were identified. We found that VPA affected gene expression in choroid plexus, excitatory neuron, immature neuron, and medial ganglionic eminence cells. The cell type-specific DEGs were enriched with mitotic nuclear division, multiple neuronal functions, and response to type I interferon and virus.
Project description:Prenatal irradiation can cause neurodevelopmental defects which are characterized by a reduction in brain size (microcephaly). The underlying molecular mechanisms in humans have so far not been studied. Here, we leveraged human forebrain organoids as a model for human embryonic brain development to investigate time- and dose-dependent effects of radiation on organoid growth. For this, organoids of 14 days and 56 days old were irradiated with acute X-ray doses of 0.5 Gy or 2 Gy and compared to controls. Using bulk RNA-seq at different early (6 h and 24 h) and late (14 days) time points after irradiation, we investigated mechanisms of radiation-induced growth defects which resulted from activation of the DNA damage response (cell cycle arrest, DNA repair, apoptosis), premature differentiation and the coordinated repression of primary microcephaly genes.