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| Status |
Public on Jan 31, 2023 |
| Title |
RNA sequencing of postmortem-fibroblast derived cell lines and postmortem human Brodmann Area 9 from Cocaine Use Disorder Subjects in the University of Texas Health Science Center at Houston Brain Collection |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Introduction: Human-derived induced pluripotent stem cell (iPSC) models of brain promise to advance our understanding of neurotoxic consequences of drug use. However, how well these models recapitulate the actual genomic landscape and cell function, as well as the drug-induced alterations, remains to be established. New in vitro models of drug exposure are needed to advance our understanding of how to protect or reverse molecular changes related to substance use disorders. Methods: We engineered a novel induced pluripotent stem cell-derived model of neural progenitor cells and neurons from cultured postmortem human skin fibroblasts, and directly compared these to isogenic brain tissue from the donor source. We assessed the maturity of the cell models across differentiation from stem cells to neurons using RNA cell type and maturity deconvolution analyses as well as DNA methylation epigenetic clocks trained on adult and fetal human tissue. As proof-of-concept of this model’s utility for substance use disorder studies, we compared morphine- and cocaine-treated neurons to gene expression signatures in postmortem Opioid Use Disorder (OUD) and Cocaine Use Disorder (CUD) brains, respectively. Results: Within each human subject (N = 2, 2 clones each), brain frontal cortex epigenetic age parallels that of skin fibroblasts and closely approximates the donor’s chronological age; stem cell induction from fibroblast cells effectively sets the epigenetic clock to an embryonic age; and differentiation of stem cells to neural progenitor cells and then to neurons progressively matures the cells via DNA methylation and RNA gene expression readouts. In neurons derived from an individual who died of opioid overdose, morphine treatment induced alterations in gene expression similar to those previously observed in OUD ex-vivo brain tissue, including differential expression of the immediate early gene EGR1, which is known to be dysregulated by opioid use. Discussion: In summary, we introduce an iPSC model generated from human postmortem fibroblasts that can be directly compared to corresponding isogenic brain tissue and can be used to model perturbagen exposure such as that seen in opioid use disorder. Future studies with this and other postmortem-derived brain cellular models, including cerebral organoids, can be an invaluable tool for understanding mechanisms of drug-induced brain alterations.
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| Overall design |
RNAseq of postmortem fibroblast-derived induced pluripotent stem cells (iPSCs), Neural Progenitor Cells (NPCs) and Neurons from one subject with opioid use disorder and one subject with cocaine use disorder. Each iPSC, NPC and Neuron line has one clonal replicate, for a total of 4 lines. Also contains RNAseq from Postmortem Brain Brodmann Area 9 for 8 cocaine use disorder subjects from the UTHealth Brain Collection. Please see GEO Accession GSE182321 for control subject data used in study
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| Contributor(s) |
Mendez E, Grimm S, Stertz L, Gunaratne P, Coarfa C, Walss-Bass C |
| Citation(s) |
36873219 |
| |
| Submission date |
Jan 31, 2023 |
| Last update date |
Jun 14, 2023 |
| Contact name |
Consuelo Walss-Bass |
| E-mail(s) |
[email protected]
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| Organization name |
University of Texas Health Science Center at Houston
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| Department |
Louis A. Faillace Department of Psychiatry and Behavioral Sciences
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| Lab |
Laboratory of Consuelo Walss-Bass PhD
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| Street address |
1941 East Road Rm. 3110
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| City |
Houston |
| State/province |
Texas |
| ZIP/Postal code |
77047 |
| Country |
USA |
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| Platforms (1) |
| GPL11154 |
Illumina HiSeq 2000 (Homo sapiens) |
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| Samples (33)
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| Relations |
| BioProject |
PRJNA929897 |
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