RNAseq analysis of DPSC-derived neurons from Neurotypical control subjects and PWS subjects
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA740014 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14881654_1.fastq.gz | Fastqsanger.gz | |||
| SRR14881654_2.fastq.gz | Fastqsanger.gz | |||
| SRR14881655_1.fastq.gz | Fastqsanger.gz | |||
| SRR14881655_2.fastq.gz | Fastqsanger.gz | |||
| SRR14881656_1.fastq.gz | Fastqsanger.gz |
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Project description:A major challenge to the study and treatment of neurogenetic syndromes is the difficulty in gaining access to live neurons from individuals with these disorders. Although other sources of stem cells are currently available for differentiation into neurons, these can involve invasive procedures and be difficult or expensive to generate limiting their use on a broad scale, especially for rare syndromes which may not be well represented in the local population. Dental pulp stem cells (DPSC) are neural crest derived multipotent stem cells that reside deep the pulp of shed (baby) teeth and have the potential for broad use in the study of neurogenetic disease. Here, we use DPSC-derived neurons to investigate the transcriptional differences between neurotypical controls and Prader-Willi syndrome (PWS) conferred by uniparental disomy (UPD) versus deletion. Additionally, we investigated the increased autism incidence within the UPD genotype by sequencing PW-UPD patients with and without autism. Using this data, we defined a PWS molecular signature and discovered a global reduction in mitochondrial transcripts in the PW-UPD +ASD neurons.
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Project description:A major challenge to the study and treatment of neurogenetic syndromes is the difficulty in gaining access to live neurons from individuals with these disorders. Although other sources of stem cells are currently available for differentiation into neurons, these can involve invasive procedures and be difficult or expensive to generate limiting their use on a broad scale, especially for rare syndromes which may not be well represented in the local population. Dental pulp stem cells (DPSC) are neural crest derived multipotent stem cells that reside deep the pulp of shed (baby) teeth and have the potential for broad use in the study of neurogenetic disease. Here, we use DPSC-derived neurons to investigate the transcriptional differences between neurotypical controls, Rapid-onset obesity with hypothalamic dysfunction, hypoventilation, and autonomic dysfunction (ROHHAD) and Central congenital hypoventilation syndrome (CCHS) subjects.
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Project description:A major challenge to the study and treatment of neurogenetic syndromes is the difficulty in gaining access to live neurons from individuals with these disorders. Although other sources of stem cells are currently available for differentiation into neurons, these can involve invasive procedures and be difficult or expensive to generate limiting their use on a broad scale, especially for rare syndromes which may not be well represented in the local population. Dental pulp stem cells (DPSC) are neural crest derived multipotent stem cells that reside deep the pulp of shed (baby) teeth and have the potential for broad use in the study of neurogenetic disease. In order to investigate the characteristics of DPSC which make them a valuable resource for the study of neurogenetic syndromes we performed a set of viability, senescence and immortalization studies on control DPSC and DPSC derived neurons. We investigated the basic transport conditions and determined the maximum passage number for primary DPSCs. We then immortalized control DPSC using human telomerase reverse trancriptase (hTERT) and evaluated both neuronal differentiation potential and gene expression changes using RNAseq. Here we show that immortalized DPSC share morphological and electrophysiological properties with non-immortalized DPSC. We also show that differentiation of DPSC into neurons changes gene expression for 1305 transcripts, while immortalized neurons differ significantly in gene expression for 183 transcripts of which 94 also changed during differentiation. Taken together, these studies indicate that immortalized dental pulp derived neruons may be a new and powerful resource for the study of rare neurological disorders where patient samples are rare or difficult to obtain. RNA-seq Analysis of 3 neurotypical control DPSC, 3 DPSC derived neurons and 3 each immortalized versions of DPSC and DPSC neurons (~3 weeks post maturation)
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Project description:A major problem in studying neurogenetic syndromes is obtaining live neurons from people with these disorders. We have taken a novel approach to studying gene expression in neurons from Angelman or Dup15q syndrome subjects using dental pulp stem cells (DPSC). Shed teeth were collected to generate DPSC lines from 3 AS deletion, 3 15q Duplication and three neurotypical subjects. mRNAseq was performed on DPSC and DPSC derived neurons. We identified 20 genes in AS, 120 genes in 15q duplication and 3 genes in both groups (DPT, MED12L and AKRIC1) that were significantly different from control DPSC-neurons. Copy number correlated to gene expression for most genes across the 15q11.2-q13.1 critical region. Two thirds of the genes differentially regulated in 15q duplication were down-regulated compared to controls including the transcription factors FOXO1 and HAND2, while in AS the genes did not show a clear trend except in the 15q region. Pathway analysis revealed increased cytokine activity related genes including four genes that increased in 15q duplication samples: CCL7, CCL2, MMP2 and MMP3; while steroid hormone biosynthesis genes were slightly enriched in both 15q duplication and AS neurons. Overall there were more dramatic changes in gene expression in the 15q) duplication than AS deletion cell lines, perhaps because the mechanism of AS may be through protein targeting by UBE3A. Nonetheless, the finding of a significant increase in both HERC2 and UBE3A in 15q duplication neurons and significant decrease in these two genes in AS deletion neurons may impact the AS phenotype, at least in deletion cases.
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Project description:A major challenge to the study and treatment of neurogenetic syndromes is the difficulty in gaining access to live neurons from individuals with these disorders. Although other sources of stem cells are currently available for differentiation into neurons, these can involve invasive procedures and be difficult or expensive to generate limiting their use on a broad scale, especially for rare syndromes which may not be well represented in the local population. Dental pulp stem cells (DPSC) are neural crest derived multipotent stem cells that reside deep the pulp of shed (baby) teeth and have the potential for broad use in the study of neurogenetic disease. In order to investigate the characteristics of DPSC which make them a valuable resource for the study of neurogenetic syndromes we performed a set of viability, senescence and immortalization studies on control DPSC and DPSC derived neurons. We investigated the basic transport conditions and determined the maximum passage number for primary DPSCs. We then immortalized control DPSC using human telomerase reverse trancriptase (hTERT) and evaluated both neuronal differentiation potential and gene expression changes using RNAseq. Here we show that immortalized DPSC share morphological and electrophysiological properties with non-immortalized DPSC. We also show that differentiation of DPSC into neurons changes gene expression for 1305 transcripts, while immortalized neurons differ significantly in gene expression for 183 transcripts of which 94 also changed during differentiation. Taken together, these studies indicate that immortalized dental pulp derived neruons may be a new and powerful resource for the study of rare neurological disorders where patient samples are rare or difficult to obtain.
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