title: NRF2/long noncoding RNA ROR signaling regulates mammary stem cell expansion and protects against estrogen genotoxicity.
abstract: Long noncoding RNAs (lncRNAs) have emerged as key regulators of gene expression in embryonic stem cell (ESC) self-renewal and differentiation. In ESCs, lncRNAs are regulated at the genetic level via transcription factor binding to lncRNA gene promoters. Here we demonstrate that the key cytoprotective transcription factor NRF2 controls lncRNA expression in mammary stem cells. By profiling lncRNAs in wild-type and NRF2 knockdown mammary stem cells, we demonstrate that the lncRNA ROR, a regulator of embryonic stem cell pluripotency, is overexpressed upon NRF2 knockdown. We performed promoter analyses and examined predicted NRF2 binding elements in the ROR promoter using luciferase reporter constructs of a ROR promoter deletion series. Our studies revealed that NRF2 binds to two specific NRF2 response elements flanking the ROR promoter and that these two NRF2 response elements are equally important to suppress ROR transcription. In addition, we identified associated H3K27me3 chromatin modification and EZH2 binding at the ROR promoter that was dependent on NRF2 binding. We observed that NRF2 knockdown or ROR overexpression leads to increased stem cell self-renewal in mammary stem cells. Furthermore, we demonstrate Nrf2 regulation of the mammary stem cell population in vivo. These observations provide further evidence for the critical role of NRF2 in maintaining normal stem cell subpopulations in mammary epithelium.
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Yongshu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Jixiang
authors.initials: J
authors.lastname: Xia
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Qinglin
authors.initials: Q
authors.lastname: Li
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yao
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Eades
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Ramkishore
authors.initials: R
authors.lastname: Gernapudi
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and.
authors.forename: Nadire
authors.initials: N
authors.lastname: Duru
authors.affiliation: Department of Pharmacology & Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Kensler
authors.affiliation: From the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 and qzhou@som.umaryland.edu.
authors.forename: Qun
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry; D045744:Cell Line, Tumor; D053595:Embryonic Stem Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology; D004967:Estrogens / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006410:Hematopoiesis; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D062085:RNA, Long Noncoding; D057050:Receptor Tyrosine Kinase-like Orphan Receptors / Q000378:metabolism; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology*
chemical_list: D000975:Antioxidants; D004967:Estrogens; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; C495636:Nfe2l2 protein, mouse; D062085:RNA, Long Noncoding; D057050:Receptor Tyrosine Kinase-like Orphan Receptors
keywords: Antioxidant; Estrogen; Estrogen Metabolite; Long Noncoding RNA (long ncRNA, lncRNA); Mammary Stem Cell; Nuclear Factor 2 (Erythroid-derived 2-like Factor) (NFE2L2) (Nrf2); ROR; Stem Cells; lincRNA-ROR