Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 117
Filter
1.
Nat Commun ; 12(1): 3638, 2021 06 15.
Article in English | MEDLINE | ID: mdl-34131144

ABSTRACT

To ensure dosage compensation between the sexes, one randomly chosen X chromosome is silenced in each female cell in the process of X-chromosome inactivation (XCI). XCI is initiated during early development through upregulation of the long non-coding RNA Xist, which mediates chromosome-wide gene silencing. Cell differentiation, Xist upregulation and gene silencing are thought to be coupled at multiple levels to ensure inactivation of exactly one out of two X chromosomes. Here we perform an integrated analysis of all three processes through allele-specific single-cell RNA-sequencing. Specifically, we assess the onset of random XCI in differentiating mouse embryonic stem cells, and develop dedicated analysis approaches. By exploiting the inter-cellular heterogeneity of XCI onset, we identify putative Xist regulators. Moreover, we show that transient Xist upregulation from both X chromosomes results in biallelic gene silencing right before transitioning to the monoallelic state, confirming a prediction of the stochastic model of XCI. Finally, we show that genetic variation modulates the XCI process at multiple levels, providing a potential explanation for the long-known X-controlling element (Xce) effect, which leads to preferential inactivation of a specific X chromosome in inter-strain crosses. We thus draw a detailed picture of the different levels of regulation that govern the initiation of XCI. The experimental and computational strategies we have developed here will allow us to profile random XCI in more physiological contexts, including primary human cells in vivo.


Subject(s)
RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Up-Regulation , X Chromosome Inactivation , Alleles , Animals , Dosage Compensation, Genetic , Female , Gene Silencing , Mice , Mice, Inbred C57BL , Mouse Embryonic Stem Cells , Sequence Analysis, RNA , X Chromosome , X Chromosome Inactivation/genetics , X Chromosome Inactivation/physiology
2.
Mol Cell Biol ; 41(8): e0038220, 2021 07 23.
Article in English | MEDLINE | ID: mdl-34060915

ABSTRACT

The long noncoding RNA XIST is the master regulator for the process of X chromosome inactivation (XCI) in mammalian females. Here, we report the existence of a hitherto-uncharacterized cis regulatory element (cRE) within the first exon of human XIST, which determines the transcriptional status of XIST during the initiation and maintenance phases of XCI. In the initiation phase, pluripotency factors bind to this cRE and keep XIST repressed. In the maintenance phase of XCI, the cRE is enriched for CTCF, which activates XIST transcription. By employing a CRISPR-dCas9-KRAB-based interference strategy, we demonstrate that binding of CTCF to the newly identified cRE is critical for regulating XIST in a YY1-dependent manner. Collectively, our study uncovers the combinatorial effect of multiple transcriptional regulators influencing XIST expression during the initiation and maintenance phases of XCI.


Subject(s)
CCCTC-Binding Factor/metabolism , Embryonic Stem Cells/metabolism , RNA, Long Noncoding/genetics , X Chromosome Inactivation/physiology , CCCTC-Binding Factor/genetics , Humans , Regulatory Sequences, Nucleic Acid/genetics , Repressor Proteins/genetics , Repressor Proteins/metabolism , X Chromosome Inactivation/genetics
3.
Med Sci (Paris) ; 37(2): 152-158, 2021 Feb.
Article in French | MEDLINE | ID: mdl-33591258

ABSTRACT

The inactivation of one of the two X chromosomes of female mammals is a vital process and a paradigm for epigenetic regulations. X-inactivation is triggered, early during embryo development, by the accumulation of a peculiar noncoding RNA, XIST, which interacts with a plethora of molecular complexes and ultimately protects the coated chromosome from the expression machinery. Once installed, the inactive state is locked by multiple layers of chromatin modifications, ensuring its stable perpetuation across cell divisions. However, recent discoveries made in various model organisms urge us to revisit some of the general principles of the X-inactivation process.


TITLE: Dernières nouvelles du chromosome X - Des principes généraux nuancés. ABSTRACT: L'inactivation d'un des deux chromosomes X des femelles mammifères est un processus vital et emblématique des régulations épigénétiques. Elle est déclenchée par l'accumulation d'un ARN non codant, XIST, qui isole le chromosome concerné de la machinerie transcriptionnelle ; l'état inactif persiste ensuite de manière stable au cours des divisions cellulaires successives. Cependant, des découvertes récentes conduisent à revisiter certains principes généraux de l'inactivation du chromosome X initialement établis. Ainsi le chercheur, tout comme le poète**, est-il invité à « vingt fois sur le métier remettre son ouvrage ¼.


Subject(s)
X Chromosome Inactivation/physiology , Animals , Female , Gene Silencing/physiology , Genetics/trends , Humans , Mammals/genetics , X Chromosome/genetics
4.
Cell Rep ; 33(10): 108485, 2020 12 08.
Article in English | MEDLINE | ID: mdl-33296655

ABSTRACT

Immune responses differ between women and men, and type I interferon (IFN) responses following Toll-like receptor 7 (TLR7) stimulation are higher in women. The precise mechanisms driving these sex differences in immunity are unknown. To investigate possible genetic factors, we quantify escape from X chromosome inactivation (XCI) for TLR7 and four other genes (RPS6KA3, CYBB, BTK, and IL13RA1) at the single plasmacytoid dendritic cell (pDC) level. We observe escape from XCI for all investigated genes, leading to biallelic expression patterns. pDCs with biallelic gene expression have significantly higher mRNA levels of the respective genes. Unstimulated pDCs with biallelic TLR7 expression exhibit significantly higher IFNα/ß mRNA levels, and IFNα exposure results in significantly increased IFNα/ß protein production by pDCs. These results identify unanticipated heterogeneity in escape from XCI of several genes in pDCs and highlight the important contribution of X chromosome factors to sex differences in type I IFN responses, which might explain observed sex differences in human diseases.


Subject(s)
Dendritic Cells/metabolism , Interferon Type I/metabolism , Toll-Like Receptor 7/metabolism , Cells, Cultured , Dendritic Cells/immunology , Dendritic Cells/physiology , Female , Gene Expression/genetics , Humans , Immunity, Innate/physiology , Interferon Type I/physiology , Male , RNA, Messenger/metabolism , Sex Characteristics , Sex Factors , Signal Transduction/genetics , X Chromosome Inactivation/genetics , X Chromosome Inactivation/physiology
5.
Nat Cell Biol ; 22(9): 1116-1129, 2020 09.
Article in English | MEDLINE | ID: mdl-32807903

ABSTRACT

How allelic asymmetry is generated remains a major unsolved problem in epigenetics. Here we model the problem using X-chromosome inactivation by developing "BioRBP", an enzymatic RNA-proteomic method that enables probing of low-abundance interactions and an allelic RNA-depletion and -tagging system. We identify messenger RNA-decapping enzyme 1A (DCP1A) as a key regulator of Tsix, a noncoding RNA implicated in allelic choice through X-chromosome pairing. DCP1A controls Tsix half-life and transcription elongation. Depleting DCP1A causes accumulation of X-X pairs and perturbs the transition to monoallelic Tsix expression required for Xist upregulation. While ablating DCP1A causes hyperpairing, forcing Tsix degradation resolves pairing and enables Xist upregulation. We link pairing to allelic partitioning of CCCTC-binding factor (CTCF) and show that tethering DCP1A to one Tsix allele is sufficient to drive monoallelic Xist expression. Thus, DCP1A flips a bistable switch for the mutually exclusive determination of active and inactive Xs.


Subject(s)
Endoribonucleases/metabolism , RNA/metabolism , Trans-Activators/metabolism , X Chromosome/metabolism , Alleles , Animals , CCCTC-Binding Factor/metabolism , Cell Line , Female , Gene Expression Regulation, Developmental/physiology , Mice , Mice, Inbred C57BL , RNA, Messenger/metabolism , Transcription, Genetic/physiology , Up-Regulation/physiology , X Chromosome Inactivation/physiology
6.
Cells ; 9(4)2020 04 19.
Article in English | MEDLINE | ID: mdl-32325818

ABSTRACT

X-chromosome inactivation (XCI) is a developmental process that aims to equalize the dosage of X-linked gene products between XY males and XX females in eutherian mammals. In female mouse embryos, paternal XCI is initiated at the 4-cell stage; however, the X chromosome is reactivated in the inner cell mass cells of blastocysts, and random XCI is subsequently initiated in epiblast cells. However, recent findings show that the patterns of XCI are not conserved among mammals. In this study, we used quantitative RT-PCR and RNA in situ hybridization combined with immunofluorescence to investigate the pattern of XCI during bovine embryo development. Expression of XIST (X-inactive specific transcript) RNA was significantly upregulated at the morula stage. For the first time, we demonstrate that XIST accumulation in bovine embryos starts in nuclei of female morulae, but its colocalization with histone H3 lysine 27 trimethylation was first detected in day 7 blastocysts. Both in the inner cell mass and in putative epiblast precursors, we observed a proportion of cells with XIST RNA and H3K27me3 colocalization. Surprisingly, the onset of XCI did not lead to a global downregulation of X-linked genes, even in day 9 blastocysts. Together, our findings confirm that diverse patterns of XCI initiation exist among developing mammalian embryos.


Subject(s)
Embryonic Development/genetics , Embryonic Development/physiology , Gene Expression Regulation, Developmental/genetics , X Chromosome Inactivation/physiology , Animals , Blastocyst/metabolism , Cattle , DNA Methylation , Genomic Imprinting/genetics , Histones/metabolism , Humans , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism
7.
Nat Commun ; 11(1): 764, 2020 02 07.
Article in English | MEDLINE | ID: mdl-32034154

ABSTRACT

Our understanding of the signalling pathways regulating early human development is limited, despite their fundamental biological importance. Here, we mine transcriptomics datasets to investigate signalling in the human embryo and identify expression for the insulin and insulin growth factor 1 (IGF1) receptors, along with IGF1 ligand. Consequently, we generate a minimal chemically-defined culture medium in which IGF1 together with Activin maintain self-renewal in the absence of fibroblast growth factor (FGF) signalling. Under these conditions, we derive several pluripotent stem cell lines that express pluripotency-associated genes, retain high viability and a normal karyotype, and can be genetically modified or differentiated into multiple cell lineages. We also identify active phosphoinositide 3-kinase (PI3K)/AKT/mTOR signalling in early human embryos, and in both primed and naïve pluripotent culture conditions. This demonstrates that signalling insights from human blastocysts can be used to define culture conditions that more closely recapitulate the embryonic niche.


Subject(s)
Cell Self Renewal/physiology , Human Embryonic Stem Cells/metabolism , Insulin-Like Growth Factor I/metabolism , Activins/metabolism , Animals , Blastocyst/metabolism , Cell Differentiation/drug effects , Cells, Cultured , Coculture Techniques , Culture Media/chemistry , Culture Media/metabolism , Culture Media/pharmacology , Endoderm/cytology , Endoderm/metabolism , Extraembryonic Membranes/cytology , Extraembryonic Membranes/metabolism , Fibroblasts/cytology , Gene Expression Regulation, Developmental , Human Embryonic Stem Cells/cytology , Humans , Induced Pluripotent Stem Cells/cytology , Induced Pluripotent Stem Cells/drug effects , Induced Pluripotent Stem Cells/physiology , Mice , Phosphatidylinositol 3-Kinases/metabolism , Receptor, IGF Type 1/metabolism , Signal Transduction , TOR Serine-Threonine Kinases/metabolism , Transcriptome , X Chromosome Inactivation/physiology
8.
FASEB J ; 34(1): 691-705, 2020 01.
Article in English | MEDLINE | ID: mdl-31914626

ABSTRACT

The inner cell mass (ICM) in blastocyst is the origin of all somatic and germ cells in mammals and pluripotent stem cells (PSCs) in vitro. As the conserved principles between pig and human, here we performed comprehensive single-cell RNA-seq for porcine early embryos from oocyte to early blastocyst (EB). We show the specification of the ICM and trophectoderm in morula and the molecular signature of the precursors. We demonstrate the existence of naïve pluripotency signature in morula and ICM of EB, and the specific pluripotent genes and the activity of signalling pathways highlight the characteristics of the naïve pluripotency. We observe the absence of dosage compensation with respect to X-chromosome (XC) in morula, and incomplete dosage compensation in the EB. However, the dynamics of dosage compensation may be independent of the expression of XIST induced XC inactivation. Our study describes molecular landmarks of embryogenesis in pig that will provide a better strategy for derivation of porcine PSCs and improve research in regenerative medicine.


Subject(s)
Blastocyst/cytology , Cell Lineage , Gene Expression Regulation, Developmental/genetics , Germ Layers/cytology , Oocytes/cytology , Animals , Gene Expression Profiling/methods , Germ Cells/cytology , Pluripotent Stem Cells/cytology , Swine , X Chromosome Inactivation/physiology
9.
Essays Biochem ; 63(6): 663-676, 2019 12 20.
Article in English | MEDLINE | ID: mdl-31782494

ABSTRACT

Monoallelic gene expression occurs in diploid cells when only one of the two alleles of a gene is active. There are three main classes of genes that display monoallelic expression in mammalian genomes: (1) imprinted genes that are monoallelically expressed in a parent-of-origin dependent manner; (2) X-linked genes that undergo random X-chromosome inactivation in female cells; (3) random monoallelically expressed single and clustered genes located on autosomes. The heritability of monoallelic expression patterns during cell divisions implies that epigenetic mechanisms are involved in the cellular memory of these expression states. Among these, methylation of CpG sites on DNA is one of the best described modification to explain somatic inheritance. Here, we discuss the relevance of DNA methylation for the establishment and maintenance of monoallelic expression patterns among these three groups of genes, and how this is intrinsically linked to development and cellular states.


Subject(s)
Alleles , DNA Methylation , DNA/metabolism , Epigenesis, Genetic/physiology , Gene Expression/physiology , Animals , DNA/genetics , DNA Methylation/physiology , Female , Genomic Imprinting/physiology , Humans , X Chromosome Inactivation/physiology
10.
J Reprod Dev ; 65(6): 533-539, 2019 Dec 18.
Article in English | MEDLINE | ID: mdl-31631092

ABSTRACT

Xist is an X-linked ribonucleic acid (RNA) gene responsible for the cis induction of X chromosome inactivation (XCI). In cloned mammalian embryos, Xist is ectopically activated at the morula to blastocyst stage on the X chromosome that is supposed to be active, thus resulting in abnormal XCI. Suppression of erroneous Xist expression by injecting small interfering RNA (siRNA) remarkably increased the developmental efficiency of cloned male mouse embryos by approximately 10-fold. However, injection of anti-Xist siRNA resulted in only a slight increase in the developmental ability of injected cloned male pig embryos because the blocking effect of the injected siRNA was not maintained beyond the morula stage, which is 5 days post-activation. To develop a more effective approach for suppressing the ectopic expression of Xist in cloned pig embryos, we compared the silencing effect of short hairpin RNA (shRNA) and siRNA on Xist expression and the effects of these two Xist knockdown methods on the developmental competence of cloned male pig embryos. Results indicated that an shRNA-based RNA interference (RNAi) has a longer blocking effect on Xist expression than an siRNA-mediated RNAi. Injection of anti-Xist shRNA plasmid into two-cell-stage cloned male pig embryos effectively suppressed Xist expression, rescued XCI at the blastocyst stage, and improved the in vitro developmental ability of injected cloned embryos. These positive effects, however, were not observed in cloned male pig embryos injected with anti-Xist siRNA. This study demonstrates that vector-based rather than siRNA-mediated RNAi of Xist expression can be employed to improve pig cloning efficiency.


Subject(s)
Cloning, Organism/methods , Embryonic Development , RNA Interference/physiology , RNA, Long Noncoding/genetics , RNA, Small Interfering/genetics , X Chromosome Inactivation/physiology , Animals , Animals, Genetically Modified , Cells, Cultured , Embryo, Mammalian , Embryonic Development/genetics , Female , Gene Expression Regulation, Developmental , Gene Knockdown Techniques/methods , Gene Knockdown Techniques/veterinary , Genetic Vectors , Male , Nuclear Transfer Techniques , RNA, Small Interfering/pharmacology , Swine/embryology , Swine/genetics
11.
Article in English | MEDLINE | ID: mdl-31450876

ABSTRACT

Rett syndrome (RTT) is a neurodevelopmental disorder with a genetic basis that is associated with the mutation of the X-linked methyl-CpG binding protein 2 (MECP2) gene in approximately 90% of patients. RTT is characterized by a brief period of normal development followed by loss of acquired skills and evolution towards impairment of brain and motor functions and multi-organ dysfunction. Originally, RTT was considered lethal in males as it has an X-linked dominant inheritance. However, although this syndrome has a higher incidence in females, rare cases are also documented in males. Here, we describe the case of an 11-year-old male patient with a microduplication MECP2 Xq28. Our patient is currently living, while his older brother with the same mutation died at the age of 9 years. We showed that the role of MECP2 as an epigenetic modulator and the X-chromosome inactivation pattern can explain the lethal clinical form of the older brother with the same microduplication MECP2 Xq28 presented by our patient who is still alive. Given the limited case history of RTT in males, further studies are needed to better characterize this syndrome in males and consequently improve the currently available therapeutic strategies.


Subject(s)
Methyl-CpG-Binding Protein 2/genetics , Rett Syndrome/genetics , X Chromosome Inactivation/genetics , Child , Dosage Compensation, Genetic , Humans , Male , Methyl-CpG-Binding Protein 2/physiology , Mutation , Patient Outcome Assessment , Siblings , X Chromosome Inactivation/physiology
13.
EMBO J ; 38(12)2019 06 17.
Article in English | MEDLINE | ID: mdl-31088843

ABSTRACT

Human pluripotent stem cells (hPSCs) are being increasingly utilized worldwide in investigating human development, and modeling and discovering therapies for a wide range of diseases as well as a source for cellular therapy. Yet, since the first isolation of human embryonic stem cells (hESCs) 20 years ago, followed by the successful reprogramming of human-induced pluripotent stem cells (hiPSCs) 10 years later, various studies shed light on abnormalities that sometimes accumulate in these cells in vitro Whereas genetic aberrations are well documented, epigenetic alterations are not as thoroughly discussed. In this review, we highlight frequent epigenetic aberrations found in hPSCs, including alterations in DNA methylation patterns, parental imprinting, and X chromosome inactivation. We discuss the potential origins of these abnormalities in hESCs and hiPSCs, survey the different methods for detecting them, and elaborate on their potential consequences for the different utilities of hPSCs.


Subject(s)
Epigenesis, Genetic/physiology , Pluripotent Stem Cells/physiology , Cell Differentiation/genetics , Cellular Reprogramming/genetics , DNA Methylation/physiology , Genomic Imprinting/genetics , Humans , Induced Pluripotent Stem Cells/physiology , X Chromosome Inactivation/physiology
14.
Prenat Diagn ; 39(8): 603-608, 2019 07.
Article in English | MEDLINE | ID: mdl-31069818

ABSTRACT

OBJECTIVE: To predict the risk of dystrophinopathy in fetal carriers of dystrophin gene (DMD) mutations. METHODS: Twenty-three pregnant women, with a total of 25 female fetuses carrying DMD mutations, were recruited. Among them, 13 pregnant women who participated in this study were only used to analyse the incidence of induced abortion after fetuses were diagnosed as dystrophinopathy carriers. Eleven fetal carriers from 10 pregnant women were tested to analyse X-chromosome inactivation (XCI) using amniocytes to assess the risk of dystrophinopathy. Follow-ups were conducted on all cases. RESULTS: Approximately one-third of fetuses were aborted before assessing the risk of dystrophinopathy. XCI analysis of amniocytes showed that 10 fetuses had random XCI patterns, and one fetus exhibited a highly skewed XCI pattern (100:0) with primary expression of the maternal X chromosome that carried the mutant allele. These 11 fetal carriers were born, and follow-up showed that the girl who showed the skewed XCI pattern as a fetus was diagnosed with Duchenne muscular dystrophy (DMD) at the age of four. The others did not present with dystrophinopathy-associated symptoms. CONCLUSIONS: XCI was significantly implicated in symptomatic female carriers of dystrophinopathies, and XCI pattern analysis of amniocytes may be useful in predicting the risk of dystrophinopathy in fetal carriers.


Subject(s)
Amnion/metabolism , Dystrophin/genetics , Fetus/metabolism , Muscular Dystrophy, Duchenne/diagnosis , X Chromosome Inactivation/physiology , Abortion, Induced/statistics & numerical data , Adult , Amnion/pathology , Cohort Studies , Female , Genetic Testing , Heterozygote , Humans , Incidence , Infant, Newborn , Male , Muscular Dystrophy, Duchenne/epidemiology , Muscular Dystrophy, Duchenne/genetics , Muscular Dystrophy, Duchenne/pathology , Mutation , Pedigree , Phenotype , Pregnancy , Prenatal Diagnosis/methods , Prognosis , Risk Factors
15.
Sci Rep ; 9(1): 6068, 2019 04 15.
Article in English | MEDLINE | ID: mdl-30988473

ABSTRACT

X chromosome inactivation (XCI) is a mammalian specific, developmentally regulated process relying on several mechanisms including antisense transcription, non-coding RNA-mediated silencing, and recruitment of chromatin remodeling complexes. In vitro modeling of XCI, through differentiation of embryonic stem cells (ESCs), provides a powerful tool to study the dynamics of XCI, overcoming the need for embryos, and facilitating genetic modification of key regulatory players. However, to date, robust initiation of XCI in vitro has been mostly limited to mouse pluripotent stem cells. Here, we adapted existing protocols to establish a novel monolayer differentiation protocol for rat ESCs to study XCI. We show that differentiating rat ESCs properly downregulate pluripotency factor genes, and present female specific Xist RNA accumulation and silencing of X-linked genes. We also demonstrate that RNF12 seems to be an important player in regulation of initiation of XCI in rat, acting as an Xist activator. Our work provides the basis to investigate the mechanisms directing the XCI process in a model organism different from the mouse.


Subject(s)
Cell Differentiation , Embryonic Stem Cells/physiology , RNA, Long Noncoding/metabolism , Ubiquitin-Protein Ligases/physiology , X Chromosome Inactivation/physiology , Animals , Cells, Cultured , Embryo, Mammalian , Female , Male , Models, Animal , Primary Cell Culture , Rats
16.
Nat Commun ; 10(1): 500, 2019 01 30.
Article in English | MEDLINE | ID: mdl-30700715

ABSTRACT

High-resolution molecular programmes delineating the cellular foundations of mammalian embryogenesis have emerged recently. Similar analysis of human embryos is limited to pre-implantation stages, since early post-implantation embryos are largely inaccessible. Notwithstanding, we previously suggested conserved principles of pig and human early development. For further insight on pluripotent states and lineage delineation, we analysed pig embryos at single cell resolution. Here we show progressive segregation of inner cell mass and trophectoderm in early blastocysts, and of epiblast and hypoblast in late blastocysts. We show that following an emergent short naive pluripotent signature in early embryos, there is a protracted appearance of a primed signature in advanced embryonic stages. Dosage compensation with respect to the X-chromosome in females is attained via X-inactivation in late epiblasts. Detailed human-pig comparison is a basis towards comprehending early human development and a foundation for further studies of human pluripotent stem cell differentiation in pig interspecies chimeras.


Subject(s)
Single-Cell Analysis/methods , X Chromosome/metabolism , Animals , Cell Differentiation/physiology , Female , Gastrulation/physiology , Gene Expression Regulation, Developmental , Germ Layers/metabolism , Humans , Swine , X Chromosome Inactivation/physiology
17.
Cell ; 176(1-2): 182-197.e23, 2019 01 10.
Article in English | MEDLINE | ID: mdl-30595450

ABSTRACT

During development, the precise relationships between transcription and chromatin modifications often remain unclear. We use the X chromosome inactivation (XCI) paradigm to explore the implication of chromatin changes in gene silencing. Using female mouse embryonic stem cells, we initiate XCI by inducing Xist and then monitor the temporal changes in transcription and chromatin by allele-specific profiling. This reveals histone deacetylation and H2AK119 ubiquitination as the earliest chromatin alterations during XCI. We show that HDAC3 is pre-bound on the X chromosome and that, upon Xist coating, its activity is required for efficient gene silencing. We also reveal that first PRC1-associated H2AK119Ub and then PRC2-associated H3K27me3 accumulate initially at large intergenic domains that can then spread into genes only in the context of histone deacetylation and gene silencing. Our results reveal the hierarchy of chromatin events during the initiation of XCI and identify key roles for chromatin in the early steps of transcriptional silencing.


Subject(s)
Chromatin/metabolism , X Chromosome Inactivation/genetics , X Chromosome Inactivation/physiology , Acetylation , Animals , Chromatin/genetics , Embryonic Stem Cells , Epigenomics/methods , Female , Gene Silencing , Histone Deacetylases/metabolism , Histones/metabolism , Mice , Polycomb-Group Proteins/metabolism , Protein Processing, Post-Translational , RNA, Long Noncoding/metabolism , Transcription, Genetic , Ubiquitination , X Chromosome/metabolism
18.
Am J Med Genet B Neuropsychiatr Genet ; 180(6): 415-427, 2019 09.
Article in English | MEDLINE | ID: mdl-30537437

ABSTRACT

Co-morbid chronic musculoskeletal pain (CMSP) and posttraumatic stress symptoms (PTSS) are frequent sequelae of motor vehicle collision, are associated with greater disability than either outcome alone, and are more prevalent in women than men. In the current study we assessed for evidence that gene transcripts originating from the X chromosome contribute to sex differences in vulnerability to CMSP and PTSS after motor vehicle collision. Nested samples were drawn from a longitudinal study of African American individuals, and CMSP (0-10 numeric rating scale) and PTSS (impact of events scale, revised) outcomes were assessed 6 months following motor vehicle collision. Blood RNA were sequenced (n = 101) and the relationship between X chromosome mRNA expression levels and co-morbid CMSP and PTSS outcomes was evaluated using logistic regression analyses. A disproportionate number of peritraumatic X chromosome mRNA predicting CMSP and PTSS in women were genes previously found to escape X chromosome inactivation (11/40, z = -2.9, p = .004). Secondary analyses assessing gene ontology relationships between these genes identified an enrichment in genes known to influence neuronal plasticity. Further, the relationship of expression of two critical regulators of X chromosome inactivation, X-inactive specific transcript (XIST) and Yin Yang 1 (YY1), was different in women developing CMSP and PTSS. Together, these data suggest that X chromosome genes that escape inactivation may contribute to sex differences in vulnerability to CMSP and PTSS after motor vehicle collision.


Subject(s)
Musculoskeletal Pain/genetics , Stress Disorders, Post-Traumatic/genetics , X Chromosome Inactivation/genetics , Accidents, Traffic/psychology , Adult , Black or African American , Chromosomes, Human, X/genetics , Chromosomes, Human, X/physiology , Comorbidity , Female , Gene Expression Regulation/genetics , Humans , Longitudinal Studies , Middle Aged , Prevalence , X Chromosome Inactivation/physiology
19.
Hum Mol Genet ; 28(8): 1331-1342, 2019 04 15.
Article in English | MEDLINE | ID: mdl-30576442

ABSTRACT

X chromosome inactivation (XCI) is a key epigenetic gene expression regulatory process, which may play a role in women's cancer. In particular tissues, some genes are known to escape XCI, yet patterns of XCI in ovarian cancer (OC) and their clinical associations are largely unknown. To examine XCI in OC, we integrated germline genotype with tumor copy number, gene expression and DNA methylation information from 99 OC patients. Approximately 10% of genes showed different XCI status (either escaping or being subject to XCI) compared with the studies of other tissues. Many of these genes are known oncogenes or tumor suppressors (e.g. DDX3X, TRAPPC2 and TCEANC). We also observed strong association between cis promoter DNA methylation and allele-specific expression imbalance (P = 2.0 × 10-10). Cluster analyses of the integrated data identified two molecular subgroups of OC patients representing those with regulated (N = 47) and dysregulated (N = 52) XCI. This XCI cluster membership was associated with expression of X inactive specific transcript (P = 0.002), a known driver of XCI, as well as age, grade, stage, tumor histology and extent of residual disease following surgical debulking. Patients with dysregulated XCI (N = 52) had shorter time to recurrence (HR = 2.34, P = 0.001) and overall survival time (HR = 1.87, P = 0.02) than those with regulated XCI, although results were attenuated after covariate adjustment. Similar findings were observed when restricted to high-grade serous tumors. We found evidence of a unique OC XCI profile, suggesting that XCI may play an important role in OC biology. Additional studies to examine somatic changes with paired tumor-normal tissue are needed.


Subject(s)
Carcinoma, Ovarian Epithelial/genetics , Genes, X-Linked/genetics , X Chromosome Inactivation/physiology , Aged , Alleles , Carcinoma, Ovarian Epithelial/metabolism , Chromosomes, Human, X/genetics , Cluster Analysis , DNA Methylation/genetics , Epigenesis, Genetic/genetics , Female , Gene Expression Regulation/genetics , Gene Frequency/genetics , Genetic Association Studies/methods , Genotype , Humans , Middle Aged , Ovarian Neoplasms/genetics , Promoter Regions, Genetic/genetics , RNA, Long Noncoding , Transcription Factors/genetics , X Chromosome Inactivation/genetics
20.
Chromosome Res ; 26(3): 179-189, 2018 09.
Article in English | MEDLINE | ID: mdl-29679205

ABSTRACT

In female mammals, each cell silences one X chromosome by converting it into transcriptionally inert heterochromatin. The inactivation is concomitant with epigenetic changes including methylation of specific histone residues and incorporation of macroH2A. Such epigenetic changes may exert influence on the positioning of the inactive X chromosome (Xi) within the nucleus beyond the level of chromatin structure. However, the dynamic positioning of the inactive X chromosome during cell cycle remains unclear. Here, we show that H3K27me3 is a cell-cycle-independent marker for the inactivated X chromosomes in WI38 cells. By utilizing this marker, three types of Xi locations in the nuclei are classified, which are envelope position (associated with envelope), mid-position (between the envelope and nucleolus), and nucleolus position (associated with the nucleolus). Moreover, serial-section analysis revealed that the inactive X chromosomes in the mid-position appear to be sparser and less condensed than those associated with the nuclear envelope or nucleolus. During the transition from G0 to G1 phase, the inactive X chromosomes tend to move from the envelope position to the nucleolus position in WI38 cells. Our results imply a role of chromosome positioning in maintaining the organization of the inactive X chromosomes in different cell phases.


Subject(s)
Chromosomes, Mammalian/metabolism , G1 Phase/physiology , Resting Phase, Cell Cycle/physiology , X Chromosome Inactivation/physiology , X Chromosome/metabolism , Animals , Cell Line , Female , Mice
SELECTION OF CITATIONS
SEARCH DETAIL
...