ATF3 deficiency impairs the proliferative–secretory phase transition and decidualization in RIF patients

AbstractDecidualization is a complex process involving cellular proliferation and differentiation of the endometrial stroma and is required to establish and support pregnancy. Dysregulated decidualization has been reported to be a critical cause of recurrent implantation failure (RIF). In this study, we found that Activating transcription factor 3 (ATF3) expression was significantly downregulated in the endometrium of RIF patients. Knockdown of ATF3 in human endometrium stromal cells (hESCs) hampers decidualization, while overexpression could trigger the expression of decidual marker genes, and ameliorate the decidualization of hESCs from RIF patients. Mechanistically, ATF3 promotes decidualization by upregulating FOXO1 via suppressing miR-135b expression. In addition, the endometrium of RIF patients was hyperproliferative, while overexpression of ATF3 inhibited the proliferation of hESCs through CDKN1A. These data demonstrate the critical roles of endometrial ATF3 in regulating decidualization and proliferation, and dysregulation of ATF3 in the endometrium may be a novel cause of RIF and therefore represent a potential therapeutic target for RIF.

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Progesterone Receptor Transcriptome and Cistrome in Decidualized Human Endometrial Stromal Cells

Endocrinology ◽

10.1210/en.2014-1566 ◽

2015 ◽

Vol 156 (6) ◽

pp. 2239-2253 ◽

Cited By ~ 41

Author(s):

Erik C. Mazur ◽

Yasmin M. Vasquez ◽

Xilong Li ◽

Ramakrishna Kommagani ◽

Lichun Jiang ◽

...

Keyword(s):

Progesterone Receptor ◽

Stromal Cells ◽

Cellular Proliferation ◽

Direct Interaction ◽

Marker Genes ◽

Regulation Of Transcription ◽

Activator Protein 1 ◽

Endometrial Stromal Cells ◽

Complex Process

Abstract Decidualization is a complex process involving cellular proliferation and differentiation of the endometrial stroma that is required to establish and support pregnancy. Progesterone acting via its nuclear receptor, the progesterone receptor (PGR), is a critical regulator of decidualization and is known to interact with certain members of the activator protein-1 (AP-1) family in the regulation of transcription. In this study, we identified the cistrome and transcriptome of PGR and identified the AP-1 factors FOSL2 and JUN to be regulated by PGR and important in the decidualization process. Direct targets of PGR were identified by integrating gene expression data from RNA sequencing with the whole-genome binding profile of PGR determined by chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) in primary human endometrial stromal cells exposed to 17β-estradiol, medroxyprogesterone acetate, and cAMP to promote in vitro decidualization. Ablation of FOSL2 and JUN attenuates the induction of 2 decidual marker genes, IGFBP1 and PRL. ChIP-seq analysis of genomic binding revealed that FOSL2 is bound in proximity to 8586 distinct genes, including nearly 80% of genes bound by PGR. A comprehensive assessment of the PGR-dependent decidual transcriptome integrated with the genomic binding of PGR identified FOSL2 as a potentially important transcriptional coregulator of PGR via direct interaction with regulatory regions of genes actively regulated during decidualization.

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Induction of ferroptosis by ATF3 elevation alleviates cisplatin resistance in gastric cancer by restraining Nrf2/Keap1/xCT signaling

Cellular & Molecular Biology Letters ◽

10.1186/s11658-021-00271-y ◽

2021 ◽

Vol 26 (1) ◽

Author(s):

Dazhi Fu ◽

Chunxiao Wang ◽

Lei Yu ◽

Rui Yu

Keyword(s):

Cisplatin Resistance ◽

Activating Transcription Factor 3 ◽

Advanced Gastric Carcinoma ◽

Kaplan Meier ◽

Promising Therapeutic Approach ◽

Activating Transcription Factor ◽

Kaplan Meier Plotter ◽

Frequent Problem ◽

Atf3 Expression ◽

Resistant Cells

Abstract Background Currently, resistance against cisplatin (DDP) is a frequent problem for the success of advanced gastric carcinoma (GC) chemotherapy. Here, we sought to investigate the function of activating transcription factor 3 (ATF3) n GC chemoresistance. Methods Expression of ATF3 was determined in GC cell lines (MNK45, SGC7901, and BGC823) and cisplatin (DDP)-resistant cells (SGC7901/DDP and BGC823/DDP). Biological informatics was performed to analyze ATF3 expression and prognosis in GC patients. Cisplatin resistance was evaluated. Ferroptosis was detected after ATF3 transfection of cells. The underlying molecular mechanism was also investigated. Results Transcripts of ATF3 were decreased in GC cells and GC tissues. Kaplan–Meier plotter analysis revealed that ATF3 expression was positively related to the overall survival of GC patients. In particular, lower levels of ATF3 were observed in cisplatin-resistant SGC7901/DDP and BGC823/DDP relative to their parental cells. Notably, ATF3 elevation sensitized cisplatin-resistant cells to cisplatin. Mechanically, compared with parental cells, SGC7901/DDP and BGC823/DDP cells exhibited lower ferroptosis evident by lower ROS, MDA and lipid peroxidation and higher intracellular GSH levels. However, ATF3 elevated ferroptosis in SGC7901/DDP and BGC823/DDP cells. Intriguingly, ATF3 overexpression together with ferroptosis activator erastin or RSL3 treatment further enhanced ferroptosis and cisplatin resistance; however, the ferroptosis suppressor liproxstatin-1 reversed the function of ATF3 in ferroptosis and cisplatin resistance. Additionally, cisplatin-resistant cells exhibited stronger activation of Nrf2/Keap1/xCT signaling relative to parental cells, which was restrained by ATF3 up-regulation. Importantly, restoring Nrf2 signaling overturned ATF3-mediated ferroptosis and cisplatin resistance. Conclusion ATF3 may sensitize GC cells to cisplatin by induction of ferroptosis via blocking Nrf2/Keap1/xCT signaling, supporting a promising therapeutic approach for overcoming chemoresistance in GC.

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Polo-like kinase expression in normal human endometrium during the menstrual cycle

Reproduction Fertility and Development ◽

10.1071/rd00023 ◽

2000 ◽

Vol 12 (2) ◽

pp. 59 ◽

Cited By ~ 6

Author(s):

Noriyuki Takai ◽

Tami Miyazaki ◽

Isao Miyakawa ◽

Ryoji Hamanaka

Keyword(s):

Menstrual Cycle ◽

Stromal Cells ◽

Cellular Proliferation ◽

Secretory Phase ◽

Ki 67 ◽

Normal Endometrium ◽

Proliferative Phase ◽

Normal Human ◽

Late Secretory Phase

The enzyme, polo-like kinase (PLK), is a mammalian serine/threonine kinase involved in cell cycle regulation. A great deal of evidence regarding the role of PLK in the cell cycle has been obtained through studies of cultured cells, though little is known about its function or even expression in vivo. The endometrium undergoes rapid proliferation and differentiation under ovarian steroid hormone control during the 28-day cycle. Thus, normal endometrium provides an excellent model in which to study the hormone dependency of PLK expression. In the present study, we examined the features of PLK expression in 20 samples of normal human endometrium during the menstrual cycle. The expression of Ki-67 and proliferating cell nuclear antigen (PCNA) were also examined as markers of proliferation. Immunohistochemical studies showed that PLK staining was detected in the basement membrane of many endometrial glands, stromal cells, and some endothelial cells. The number of PLK-positive endometrial gland cells was significantly higher in the late proliferative phase (19.16% 4.98%) and the early secretory phase (19.28% 4.99%) than in the early proliferative phase (2.60% 2.33%) or the late secretory phase (5.76% 2.16%) (P<0.0001). PLK expression seemed to be correlated with the expression of Ki-67 and PCNA in many endometrial glands and stromal cells particularly in the late proliferative phase, reflecting a role of PLK in cellular proliferation. Nevertheless, in the early secretory phase, at which point the expression of Ki-67 and PCNA decreased in endometrial glands, PLK was strongly expressed. This finding suggests that PLK may have some post-mitotic functions in certain specialized cell types. Although the highest expression of PLK was observed in the late proliferative and the early secretory phases, the expression drastically decreased in the late secretory phase. These findings, taken together, indicate that the expression of PLK in normal endometrium fluctuates over the course of the menstrual cycle, suggesting in turn that PLK is associated with hormone-dependent cellular proliferation and that hormone functions may be involved in its regulation.

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Hmgb3 Induces the Differentiation of Uterine Stromal Cells Through Targeting Ptn

Reproductive Sciences ◽

10.1177/1933719118792098 ◽

2018 ◽

Vol 26 (7) ◽

pp. 891-899

Author(s):

Kai Wang ◽

Yun-Hou Yin ◽

Zhan-Qing Yang ◽

Hai-Fan Yu ◽

Yu-Si Wang ◽

...

Keyword(s):

Stromal Cells ◽

Messenger Rna ◽

Cellular Proliferation ◽

Physiological Role ◽

High Mobility ◽

Ovariectomized Mice ◽

Dynamic Expression ◽

Proliferation And Differentiation ◽

Reliable Marker

Uterine decidualization is crucial for placenta formation and pregnancy maintenance. Although previous studies have reported that high mobility group box 3 (Hmgb3) is involved in the regulation of cellular proliferation and differentiation, little is known regarding its physiological role in uterine decidualization. Here, in situ hybridization result exhibited a dynamic expression pattern of Hmgb3 messenger RNA (mRNA) during early gestation, and it was mainly localized to the decidua on days 6 to 8 of gestation. Consistently, elevated Hmgb3 expression was noted in the decidualizing stromal cells after intraluminal oil infusion. In uterine luminal epithelium of ovariectomized mice, estrogen induced the accumulation of Hmgb3 mRNA, which was dependent on the existence of implanting blastocyst. Simultaneously, Hmgb3 could stimulate the proliferation of uterine stromal cells and promote the expression of Prl8a2, a reliable marker for stromal cell differentiation. Further analysis evidenced that Hmgb3 might modulate the expression of pleiotropin (Ptn) in uterine stromal cells. Moreover, silencing of Ptn could impede the upregulation of Prl8a2 elicited by Hmgb3 overexpression, while overexpression of Ptn reversed the repressive effects of Hmgb3 siRNA on Prl8a2 expression. Collectively, Hmgb3 may direct uterine decidualization through targeting Ptn.

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Activating transcription factor 3: a hormone responsive gene in the etiology of hypospadias

Acta Endocrinologica ◽

10.1530/eje-07-0793 ◽

2008 ◽

Vol 158 (5) ◽

pp. 729-739 ◽

Cited By ~ 51

Author(s):

Ana Beleza-Meireles ◽

Virpi Töhönen ◽

Cilla Söderhäll ◽

Christian Schwentner ◽

Christian Radmayr ◽

...

Keyword(s):

Transcription Factor ◽

Mutation Screening ◽

Activating Transcription Factor 3 ◽

Healthy Controls ◽

Nucleotide Polymorphisms ◽

Genital Development ◽

Responsive Gene ◽

Activating Transcription Factor ◽

Atf3 Expression ◽

Transcription Factor 3

IntroductionHypospadias is a common inborn error of the genital development, whose complex etiology remains elusive. Defects of the androgen metabolism and activity have been found in a subset of boys with hypospadias. Moreover, the balance between androgens and estrogens seems to be important to the proper male genital development. Activating transcription factor 3 (ATF3), an estrogen responsive gene, has been reported to be expressed during sexual development and up-regulated in hypospadic genital skin. We investigated ATF3 as a candidate gene for hypospadias.Material and methodsGenotyping of eight-tagged single nucleotide polymorphisms (SNP)s was performed in 330 boys with hypospadias and in 380 healthy controls. Screening for mutations in ATF3 was conducted in a subset of boys with hypospadias. ATF3 expression was evaluated in the foreskin of boys with hypospadias and in healthy controls and in the human fetal genitalia by immunohistochemistry.ResultsThree common SNPs, spanning a region of about 16 kb in intron 1 of ATF3, are associated with hypospadias. These SNPs are not linked and their effects are independent. The combination of the three risk SNPs yields the highest significance. Mutation screening identified the gene variant c536A>G in one patient and c817C>T in the 3′-UTR in two other patients. ATF3 expression was evidenced in the developing male urethra.ConclusionsATF3 gene variants influence the risk of hypospadias. Its hormonal responsiveness may underlie this risk effect. But also other ATF3-dependent biological aspects, such as cell survival and death, response to stress stimuli, or the control of epithelial–mesenchymal interactions, may be of importance.

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Knockdown of RyR3 Enhances Adiponectin Expression Through an atf3-Dependent Pathway

Endocrinology ◽

10.1210/en.2012-1515 ◽

2013 ◽

Vol 154 (3) ◽

pp. 1117-1129 ◽

Cited By ~ 5

Author(s):

Shu-Huei Tsai ◽

Emily Yun-Chia Chang ◽

Yi-Cheng Chang ◽

Siow-Wey Hee ◽

Yun-Chih Tsai ◽

...

Keyword(s):

Specific Protein ◽

Activating Transcription Factor 3 ◽

Mrna Levels ◽

Mitochondrial Mass ◽

Glucose Levels ◽

Adiponectin Mrna ◽

Activating Transcription Factor ◽

Interfering Rna ◽

Dependent Pathway ◽

Atf3 Expression

Abstract Adiponectin is an important adipose-specific protein, which possesses insulin (INS)-sensitizing, antiinflammatory, and antiatherosclerotic functions. However, its regulation remains largely unknown. In this study, we identified that ryanodine receptor (RyR)3 plays an important role in the regulation of adiponectin expression. RyR3 was expressed in 3T3-L1 preadipocytes, and its level was decreased upon adipogenesis. Silencing of RyR3 expression in 3T3-L1 preadipocytes resulted in up-regulated adiponectin promoter activity, enhanced adiponectin mRNA expression, and more adiponectin protein secreted into the medium. An inverse relation between RyR3 and adiponectin mRNA levels was also observed in adipose tissues of db/db mice. In addition, knockdown of RyR3 with small interfering RNA (siRNA) in db/db mice and high-fat diet-fed obese mice increased serum adiponectin level, improved INS sensitivity, and lowered fasting glucose levels. These effects were in parallel with decreased mitochondrial Ca2+, increased mitochondrial mass, and reduced activating transcription factor 3 (atf3) expression. Overexpression of atf3 in 3T3-L1 preadipocytes blocked the effect of RyR3 silencing on adiponectin expression, indicating that an atf3-dependent pathway mediates the effect downstream of RyR3 silencing. Our data suggest that RyR3 may be a new therapeutic target for improving INS sensitivity and related metabolic disorders.

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Ozone induces pulmonary activating transcription factor 3 (ATF3) expression

The FASEB Journal ◽

10.1096/fasebj.25.1_supplement.1037.2 ◽

2011 ◽

Vol 25 (S1) ◽

Author(s):

Yuchieh Wu ◽

Wesley Konsavage ◽

Joanna Floros ◽

Jeffrey S Shenberger

Keyword(s):

Transcription Factor ◽

Activating Transcription Factor 3 ◽

Activating Transcription Factor ◽

Atf3 Expression ◽

Transcription Factor 3

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Abstract P795: Activating Transcription Factor 3 Improves Stroke Recovery Through Reduction of Neuronal Damage and Neuroinflammation

Stroke ◽

10.1161/str.52.suppl_1.p795 ◽

2021 ◽

Vol 52 (Suppl_1) ◽

Author(s):

Zhanqiang Wang ◽

Peipei Pan ◽

Rich Liang ◽

Qifeng Li ◽

Janothon Pan ◽

...

Keyword(s):

Transcription Factor ◽

Ischemic Stroke ◽

Neuronal Damage ◽

Stroke Recovery ◽

Activating Transcription Factor 3 ◽

Double Labeling ◽

Activating Transcription Factor ◽

Adhesive Removal ◽

Atf3 Expression ◽

Transcription Factor 3

Background and Purpose: Activating transcription factor 3 (ATF3), a member of the ATF/CREB family, is upregulated in the early phase of various stresses including ischemic stroke. The role of ATF3 in ischemic stroke injury has not been fully elucidated. Hypothesis: ATF3 improves stroke outcomes through reduction of neuronal damage and neuroinflammation. Methods: Permanent distal middle cerebral artery occlusion (pMCAO) was performed in 8-week-old male and female C57BL/6J (WT) and ATF3 knockout (ATF3 -/- ) mice. The sensorimotor functions were analyzed 3 days after pMCAO through adhesive removal and corner tests. Infarct volumes and injured neurons were quantified on Nissl stained and Fluoro-Jade C (FJC) stained sections. The cell-specific expression of ATF3 was analyzed by co-staining ATF3 antibody with antibodies specific to NeuN (Neuron), CD68 (microglia/macrophage) and GFAP (astrocyte). The expression of ATF3 in injured neurons was analyzed by ATF3 and FJC double labeling. Results: ATF3 expression was detected exclusively in neurons in the infarct area 1 day after pMCAO. There were a few ATF3 + CD68 + microglia/macrophages at the peri-infarct regions 2 and 3 days after pMCAO. Almost all FJC + neurons were ATF3 positive. No ATF3 expression was detected in astrocytes. Compared to WT mice, ATF3 -/- mice took longer time to remove the adhesive from their right paws (p<0.001) in adhesive removal test and more left turns in corner test (p<0.001) 3 days after pMCAO. ATF3 -/- mice also had a larger infarct volume (p = 0.014), more FJC + neurons (p=0.002) and CD68 + microglia/macrophages (p=0.003) in the peri-infarct regions than WT mice. Conclusions: Deletion of ATF3 exacerbates neuronal injury, neuroinflammation, and sensorimotor dysfunction in stroke mice, suggesting that upregulation of ATF3 at early stage of stroke improves ischemic stroke recovery through reduction of neuronal damage and neuroinflammation.

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Steroidogenic Factor 1 (NR5A1) Activates ATF3 Transcriptional Activity

International Journal of Molecular Sciences ◽

10.3390/ijms21041429 ◽

2020 ◽

Vol 21 (4) ◽

pp. 1429

Author(s):

Natsuko Emura ◽

Chiung-Min Wang ◽

William Harry Yang ◽

Wei-Hsiung Yang

Keyword(s):

Site Directed Mutagenesis ◽

Activating Transcription Factor 3 ◽

Dependent Manner ◽

Orphan Nuclear Receptor ◽

Steroidogenic Factor 1 ◽

Post Translational Modifications ◽

Endocrine Organs ◽

Activating Transcription Factor ◽

Dose Dependent ◽

Atf3 Expression

Steroidogenic Factor 1 (SF-1/NR5A1), an orphan nuclear receptor, is important for sexual differentiation and the development of multiple endocrine organs, as well as cell proliferation in cancer cells. Activating transcription factor 3 (ATF3) is a transcriptional repressor, and its expression is rapidly induced by DNA damage and oncogenic stimuli. Since both NR5A1 and ATF3 can regulate and cooperate with several transcription factors, we hypothesized that NR5A1 may interact with ATF3 and plays a functional role in cancer development. First, we found that NR5A1 physically interacts with ATF3. We further demonstrated that ATF3 expression is up-regulated by NR5A1. Moreover, the promoter activity of the ATF3 is activated by NR5A1 in a dose-dependent manner in several cell lines. By mapping the ATF3 promoter as well as the site-directed mutagenesis analysis, we provide evidence that NR5A1 response elements (−695 bp and −665 bp) are required for ATF3 expression by NR5A1. It is well known that the transcriptional activities of NR5A1 are modulated by post-translational modifications, such as small ubiquitin-related modifier (SUMO) modification and phosphorylation. Notably, we found that both SUMOylation and phosphorylation of NR5A1 play roles, at least in part, for NR5A1-mediated ATF3 expression. Overall, our results provide the first evidence of a novel relationship between NR5A1 and ATF3.

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