GeneSet Information

Tier IV GS409615 • Differentially expressed genes in methadone treated human cortical organoids from iPSC line A vs untreated_qvalue

DESCRIPTION:

Human induced pluripotent stem cell (iPSC) lines, A and B, derived from two healthy adult male individuals, were used to generate hCOs for RNA-sequencing. Methodone treatment began on Day 9 of organoid culture, the first day of the neural proliferation stage, and concluded at Day 60. Nuclease-free water was used as a vehicular control. Cortical organoids were collected 2 months (60 days) after initiating organoid culture. Each well of hCOs (15–20 organoids) was a separate biological replicate for a given treatment condition (i.e., treated or untreated). RNA was extracted from frozen organoid pellets using the Direct-Zol Miniprep Plus Kit (Zymo, Irvine, CA) according to the manufacturer’s instructions. Samples were multiplexed and sequenced on the Illumina NovaSeq 6000 S4 to produce approximately 100 million, 100 base pair, paired end reads per sample. 3 control and 3 methadone-treated samples were sequenced from cell line A, and 4 control and 4 treated samples from cell line B. Raw fastq file quality assessment and read alignment to the hg19 genome (GRCh37, RefSeq GCF_000001405.13) were performed. Significantly differentially expressed genes (DEGs) were selected based on the confident effect size of their log2(Fold Change) values at FDR<0.05. Genes presented are without cutoffs and were obtained using the GEO2R tool by GW curators (GEO accession: GSE210682).

LABEL:

DEG methadone human cortical organoids cell line A_qvalue

SCORE TYPE:

Q-Value

THRESHOLD:

<= 0.5

GENES IN THRESHOLD:

15109

DATE ADDED:

2024-12-05

DATE UPDATED:

2024-12-05

SPECIES:

AUTHORS:

Ila Dwivedi, Andrew B Caldwell, Dan Zhou, Wei Wu, Shankar Subramaniam, Gabriel G Haddad

TITLE:

Methadone alters transcriptional programs associated with synapse formation in human cortical organoids.

JOURNAL:

Translational psychiatry May 2023, Vol 13, pp. 151

ABSTRACT:

Opioid use disorder (OUD) among pregnant women has become an epidemic in the United States. Pharmacological interventions for maternal OUD most commonly involve methadone, a synthetic opioid analgesic that attenuates withdrawal symptoms and behaviors linked with drug addiction. However, evidence of methadone's ability to readily accumulate in neural tissue, and cause long-term neurocognitive sequelae, has led to concerns regarding its effect on prenatal brain development. We utilized human cortical organoid (hCO) technology to probe how this drug impacts the earliest mechanisms of cortico-genesis. Bulk mRNA sequencing of 2-month-old hCOs chronically treated with a clinically relevant dose of 1 μM methadone for 50 days revealed a robust transcriptional response to methadone associated with functional components of the synapse, the underlying extracellular matrix (ECM), and cilia. Co-expression network and predictive protein-protein interaction analyses demonstrated that these changes occurred in concert, centered around a regulatory axis of growth factors, developmental signaling pathways, and matricellular proteins (MCPs). TGFβ1 was identified as an upstream regulator of this network and appeared as part of a highly interconnected cluster of MCPs, of which thrombospondin 1 (TSP1) was most prominently downregulated and exhibited dose-dependent reductions in protein levels. These results demonstrate that methadone exposure during early cortical development alters transcriptional programs associated with synaptogenesis, and that these changes arise by functionally modulating extra-synaptic molecular mechanisms in the ECM and cilia. Our findings provide novel insight into the molecular underpinnings of methadone's putative effect on cognitive and behavioral development and a basis for improving interventions for maternal opioid addiction. PUBMED: 37147277
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Annotation Information

No sequence read archive data associated with this GeneSet.


Aptitude (D001076)
Cell Line (D002460)
United States (D014481)
Induced Pluripotent Stem Cells (D057026)
Behavior (D001519)
Substance-Related Disorders (D019966)
Biological Products (D001688)
Thrombospondin 1 (D019700)
Methadone (D008691)
Opioid-Related Disorders (D009293)
RNA, Messenger (D012333)
Base Pairing (D020029)
Proteins (D011506)
Therapeutics (D013812)
Culture (D003469)
Extracellular Matrix (D005109)
Substance Withdrawal Syndrome (D013375)
Intercellular Signaling Peptides and Proteins (D036341)
Pharmaceutical Preparations (D004364)
Tissues (D014024)
Thrombospondins (D019699)
Microscopy, Electron, Scanning Transmission (D017348)
Pluripotent Stem Cells (D039904)
Stem Cells (D013234)
Pregnant Women (D037841)
Programs (D019542)
Technology (D013672)
Organoids (D009940)
Synapses (D013569)
Analgesics (D000700)
Analgesics, Opioid (D000701)
cervical vertebra 2 (MA:0001422)
addiction (MP:0002555)
stem cell factor receptor activity (GO:0005020)
synapse assembly (GO:0007416)
extracellular region (GO:0005576)
extracellular matrix (GO:0031012)
developmental process (GO:0032502)
brain development (GO:0007420)
signaling (GO:0023052)
Alignment construction (EDAM_operation:2928)
Developmental biology resources (EDAM_topic:3064)
Pathway or network (EDAM_data:2600)
Transcriptomics (EDAM_topic:0203)
Pathways, networks and models (EDAM_topic:0602)
Oligonucleotide alignment construction (EDAM_operation:3198)
Signaling pathways (EDAM_topic:0754)
Protein analysis (EDAM_topic:0078)
nucleobase (CHEBI:18282)
zoledronic acid (CHEBI:46557)
maleate(2-) (CHEBI:30780)
methadone (CHEBI:6807)
protein polypeptide chain (CHEBI:16541)
ribonucleic acid (CHEBI:33697)
messenger RNA (CHEBI:33699)
opioid analgesic (CHEBI:35482)
analgesic (CHEBI:35480)
induced pluripotent stem cell (EFO:0004905)
treatment (EFO:0000727)
pluripotent (EFO:0002967)
obsolete_stem (EFO:0001040)
drug dependence (EFO:0003890)
obsolete_adrenal cortex (EFO:0000237)
obsolete_induced pluripotent stem cell (EFO:0005739)
obsolete_renal cortex (EFO:0005891)
rectal adenocarcinoma (EFO:0005631)
protein measurement (EFO:0004747)
biological replicate (EFO:0002091)
replicate (EFO:0000683)
vertebral bone 2 (UBERON:0001093)
life cycle stage (UBERON:0000105)
male organism (UBERON:0003101)
neural tissue (UBERON:0003714)
adult organism (UBERON:0007023)
proliferative region (UBERON:2000098)
collection of eyelashes (UBERON:0010168)
adult cerebral ganglion (UBERON:6110636)
matrix compartment (UBERON:0027368)

Gene List • 18562 Genes

Genes in threshold: 15109

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