A novel Oct4/Pou5f1-like non-coding RNA controls neural maturation and mediates developmental effects of ethanol

• Published in: Neurotoxicology and teratology Vol. 83; p. 106943
• Main Authors: Salem, Nihal A., Mahnke, Amanda H., Tseng, Alexander M., Garcia, Cadianna R., Jahromi, Hooman K., Geoffroy, Cédric G., Miranda, Rajesh C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2021
• Subjects: Animals; Argonaute Proteins - metabolism; Brain - drug effects; Brain - embryology; Brain - metabolism; Cell Differentiation - drug effects; Cells, Cultured; Disease Models, Animal; Ethanol - toxicity; FASD; Female; Fetal Alcohol Spectrum Disorders - genetics; Fetal Alcohol Spectrum Disorders - metabolism; Fetal Alcohol Spectrum Disorders - pathology; Fetal neural progenitor cells; Fetal Stem Cells - drug effects; Fetal Stem Cells - metabolism; Fetal Stem Cells - pathology; Gene Expression Regulation, Developmental - drug effects; Gene Knockdown Techniques; Humans; Mice; Mice, Inbred C57BL; miRNAs; Models, Neurological; ncRNA; Neural development; Neural Stem Cells - drug effects; Neural Stem Cells - metabolism; Neural Stem Cells - pathology; Neurogenesis - drug effects; Neurogenesis - genetics; Oct4/Pou5f1; Octamer Transcription Factor-3 - antagonists & inhibitors; Octamer Transcription Factor-3 - genetics; Octamer Transcription Factor-3 - metabolism; Pregnancy; Prenatal Exposure Delayed Effects - genetics; Prenatal Exposure Delayed Effects - metabolism; Prenatal Exposure Delayed Effects - pathology; Pseudogene; Pseudogenes; RNA, Untranslated - genetics; RNA, Untranslated - metabolism; Single-Cell Analysis; Pseudogene; Oct4/Pou5f1; miRNAs; Fetal neural progenitor cells; ncRNA; FASD; Neural development
• ISSN: 0892-0362; 1872-9738
• DOI: 10.1016/j.ntt.2020.106943

Prenatal ethanol exposure can result in loss of neural stem cells (NSCs) and decreased brain growth. Here, we assessed whether a noncoding RNA (ncRNA) related to the NSC self-renewal factor Oct4/Pou5f1, and transcribed from a processed pseudogene locus on mouse chromosome 9 (mOct4pg9), contributed to the loss of NSCs due to ethanol. Mouse fetal cortical-derived NSCs, cultured ex vivo to mimic the early neurogenic environment of the fetal telencephalon, expressed mOct4pg9 ncRNA at significantly higher levels than the parent Oct4/Pou5f1 mRNA. Ethanol exposure increased expression of mOct4pg9 ncRNA, but decreased expression of Oct4/Pou5f1. Gain- and loss-of-function analyses indicated that mOct4pg9 overexpression generally mimicked effects of ethanol exposure, resulting in increased proliferation and expression of transcripts associated with neural maturation. Moreover, mOct4pg9 associated with Ago2 and with miRNAs, including the anti-proliferative miR-328-3p, whose levels were reduced following mOct4pg9 overexpression. Finally, mOct4pg9 inhibited Oct4/Pou5f1–3’UTR-dependent protein translation. Consistent with these observations, data from single-cell transcriptome analysis showed that mOct4pg9-expressing progenitors lack Oct4/Pou5f1, but instead overexpress transcripts for increased mitosis, suggesting initiation of transit amplification. Collectively, these data suggest that the inhibitory effects of ethanol on brain development are explained, in part, by a novel ncRNA which promotes loss of NSC identity and maturation. •Oct4pg9 is a novel mouse pseudogene-encoded noncoding RNA (ncRNA), related to the gene encoding the transcription factor Oct4/Pou5f1, is expressed in neural stem cells (NSCs), and is induced by alcohol (ethanol) exposure•Oct4pg9 ncRNA and Oct4/Pou5f1 mRNA are expressed in non-overlapping sub-populations of neural stem cells.•Oct4pg9 associates with miRNA-chaperone Ago2, and with the antiproliferative miRNA, miR-328-3p.•Oct4pg9 expression results in inhibition of Oct4/Pou5f1 3’UTR-dependent translation.•Gain- and loss-of-function studies indicate that Oct4pg9 upregulation explains pro-maturation and teratogenic effects of ethanol on NSCs.