The Expression Profile of Wnt/β-Catenin Signalling Pathway Genes in Miscarriages

The lncRNA DEANR1 facilitates human endoderm differentiation by activating FOXA2 expression.

To investigate the efficacy and prognosis of acute myeloid leukemia (AML) patients with chromosome karyotype abnormalities. The clinical features and treatment responses of 91 patients with AML were collected and analyzed retrospectively. The efficacy and survival rate of the AML patients with normal and abnormal chromosome karyotype were compared. Chromosome translocations and monosomal karyotypes were the main heterogeneity of AML. There was no significant difference in complete remission rate and overall response rate between the normal and abnormal karyotype groups, but the recurrence rate was higher in abnormal karyotype group. There was no significant difference in response of AML patients received the standard "3+7 regimen" and pre-excitation chemotherapy in the treatment of normal and abnormal karyotype groups. The relapse free survival time (RFS) was longer in the normal karyotype group, but there was no significant difference in overall survival time (OS). The abnormal karyotype of AML is an independent prognostic factor, monosomal karyotype shows a poor prognosis, and the recurrence rate in AML patients with monosomal karyotype is higher.

Somatic Mutations Predict Poor Prognosis in Myelodysplastic Syndrome Patients with Normal Karyotypes

To explore the role of specific AT rich sequence binding protein 1(SATB1) and wnt/β-catenin signaling pathways in the regulation of trophoblast invasion and its effect in the pathogenesis of preeclampsia. From March 2013 to March 2014, 20 cases of human villous tissues (early pregnancy group) from women of 8-10 gestational weeks who received artificial abortion at the First Affiliated Hospital of Chongqing Medical University, 18 cases of placental tissues (mid-pregnancy group) from women of 18-20 gestational weeks who had labor induction by water bag, 20 cases of placental tissues (normal full-term group) from healthy full-term pregnancy women and 20 cases of placental tissues (preeclamptic group) from women with preeclampsia who received elective c-section in were collected. Immunohistochemical SP method was utilized to determine the position of SATB1 and beta-catenin in villous tissues or placental tissues. Western blot was performed to analyze the expression level of SATB1 and beta-catenin in villous tissues or placental tissues. Immunofluorescence assay was used to determine the location of SATB1 and β-catenin in HTR8/SVneo cells. Western blot was performed to detect the expression level of SATB1 and beta-catenin in HTR8/SVneo cells cultured in normoxia and hypoxia reoxygenation (H/R) condition. Co-Immunoprecipitation detection was used to evaluate the interaction between SATB1 and β-catenin in placental tissues in preeclamptic group and HTR8/SVneo cells in H/R group. Gelatin zymography analysis was used to measure the activity of matrix metalloproteinases (MMP)-2 and 9 in placental tissues from preeclamptic group and HTR8/SVneo cells in H/R group. (1) In the normal full-term group, rare syncytiotrophoblastic nodule, less fibrinoid necrosis and abundant numbers of capillary could be observed in placental tissues. In comparison, there were obvious vacuolation in the cytoblast of syncytiotrophoblast, rich fibrinoid necrosis and poor numbers of villous capillary in placental tissues from preeclamptic group. (2) SATB1 could be found by immunochemical staining in placenta or villous tissues from all the groups. The staining intensity of SATB1 were more weakening in preeclamptic group than in the normal full-term group. (3) β-catenin could be found by immunochemical staining in placenta or villous tissues from all the groups. The staining intensity of β-catenin were more weakening in preeclamptic group than in the normal full-term group. (4) The protein expression levels of SATB1 in early pregnancy group, mid-pregnancy group, normal full-term group and preeclamptic group were 0.300 ± 0.009, 0.271±0.015, 0.238±0.018 and 0.153±0.007, respectively. The protein levels of β-catenin among the above groups were 0.743±0.041, 0.648±0.021, 0.549±0.069 and 0.269±0.047, respectively. Both the expression of SATB1 and β-catenin protein were significant decreased in placental tissues from preeclamptic group compared with the other three groups. (5) The SATB1 and β-catenin protein was located in nucleus of trophoblast and a small amount was in the cytoplasm. The fluorescence intensity of both SATB1 and β-catenin in the H/R group were significantly decreasing when compared to the normoxia group. (6) HTR8/SVneo cells in H/R group showed a significant decrease in both SATB1 and β-catenin protein levels when compared to the normoxia group. The protein level of SATB1 in the normoxia group was 0.213±0.005, while was 0.083±0.021 in the H/R group. The protein level of β-catenin in the normoxia group was 0.797±0.081, and was 0.543±0.131 in the H/R group. (7) There was an interaction between SATB1 and β-catenin in placental tissues from the preeclamptic group and HTR8/SVneo cells exposed by H/R. (8) The enzymatic activity of MMP-2 and MMP-9 protein were decreased significantly in placental tissues from the preeclamptic group (2.251±0.310, 1.447±0.102, respectively) when compared to the normal full-term group (7.098±0.451, 5.502±0.197, respectively). MMP-2 and MMP-9 were significantly decreased in the H/R group (0.471±0.104, 0.297±0.103, respectively) when compared to the normoxia group (0.842±0.209, 0.595±0.100, respectively). The expression of SATB1 decreased in the placenta of preeclampsia. This may influence the activity of MMP-2 and 9 by regulating Wnt/β-catenin signaling pathways, affect trophoblast invasion and eventually result in preeclampsia.

BackgroundMetabolic homeostasis is closely related to early impairment of cell fate determination and embryo development. The protein kinase mechanistic target of rapamycin (mTOR) is a key regulator of cellular metabolism in the body. Inhibition of mTOR signaling in early embryo causes postimplantation development failure, yet the mechanisms are still poorly understood.MethodsPregnancy mice and preimplantation mouse embryo were treated with mTOR inhibitor in vivo and in vitro respectively, and subsequently examined the blastocyst formation, implantation, and post-implantation development. We used immunofluorescence staining, RNA-Seq smart2, and genome-wide bisulfite sequencing technologies to investigate the impact of mTOR inhibitors on the quality, cell fate determination, and molecular alterations in developing embryos.ResultsWe showed mTOR suppression during preimplantation decreases the rate of blastocyst formation and the competency of implantation, impairs the post implantation embryonic development. We discovered that blocking mTOR signaling negatively affected the transformation of 8-cell embryos into blastocysts and caused various deficiencies in blastocyst quality. These included problems with compromised trophectoderm cell differentiation, as well as disruptions in cell fate specification. mTOR suppression significantly affected the transcription and DNA methylation of embryos. Treatment with mTOR inhibitors increase lysosomal activation and disrupts the organization and dynamics of the actin cytoskeleton in blastocysts.ConclusionsThese results demonstrate that mTOR plays a crucial role in 8-cell to blastocyst transition and safeguards embryo quality during early embryo development.

Congenital heart diseases are the common disease in birth defects .There are 120000 to 200000 children born with congenital heart disease in China each year , most of which are complex congenital heart diseases , seriously harming the health of the newborn , and bring heavy burden to commu-nity and the patient＇s family .The pathogenesis of congenital heart disease is complex , mostly of which due to the co-effects of environmental and genetic factors .Many patients get congenital heart diseases due to the combined action of multiple genes and signaling pathways , such as TGF-β, Nodal , MAPK , Wnt , Notch and homocysteine methionine signaling pathways which are closely associated with the cardiac devel -opment , and any abnormality in these genes or signal path ways may lead to congenital heart disease .In this paper , we summarizd the roles of various signaling pathways and their related genes in heart development , and how changes or abnormalities in these signaling pathways and genes caused congenital heart disease . Key words: Congenital heart disease; Tetralogy of fallot; Signaling pathway; Genetic fac-tors; Gene regulation

Under acute or chronic stresses, the adult heart undergoes a remodeling process that involves cardiomyocyte hypertrophy accompanied by apoptosis, necrosis, and fibrosis that lead to impaired cardiac contractility. The role of endogenous regeneration in this process is currently under investigation. Sustained deleterious stimuli will lead to a decompensated form of hypertrophy often culminating in heart failure.1 This form of hypertrophy is often referred to as “maladaptive.” When dealing with hypertrophy, it appears important to distinguish between the term being used on the cellular and molecular level (enlargement of individual cardiomyocytes and re-expression of fetal/embryonic genes) and the organ level (increased heart weight, left ventricular wall thickness, and functional diastolic and systolic impairment). In our view, these processes are certainly linked but not identical. Hypertrophy on the organ levels summarizes several independent cellular and molecular processes (see below), where cardiomyocyte growth is not necessarily the most important. Independent of its origin, cardiac hypertrophy is associated with alterations in cardiac geometry, mass, architecture, and function controlled by a complex network of interconnected and abundant signal-transduction pathways.2 New signaling molecules are emerging as possible targets to specifically attenuate maladaptive hypertrophy. Pathological, stress-induced growth of cardiomyocytes was shown to depend on Wnt/β-catenin nuclear signaling rather than its adhesive function in cell adhesion. However, the specificity of the cell type and the molecular mechanisms governing the Wnt signaling–dependent changes are currently unknown.3 In this issue of the Hypertension , the study by Malekar et al4 provides new evidences concerning the …

RORγt plays vital roles in development and differentiation of multiple immune cell types with pleiotropic functions, including thymocytes, LTi cells, ILC3, γδ T-cells and Th17 cells. Moreover, RORγ, the isoform of RORγt, was reported to promote cancer cell survival in castration-resistant prostate cancer. RORγ is expressed in tissues like muscle, liver and brain, but its function remains unclear. To address the mechanisms by which RORγt/RORγ contributes to diverse cell fates and differentiation programs, we are exploring RORγt post-translational modifications, RORγt/RORγ-associated protein complexes and downstream signals. To identify RORγt/RORγ-associated protein complexes, we generated knock-in mice expressing Strep-tagged RORγt/RORγ, which had IL-17A inducing activity similar to that of non-tagged RORγt in Th17 cells polarized in vitro and allowed for highly specific isolation of protein complexes. Strep-tagged RORγt proteins will be precipitated from thymocytes and polarized pathogenic or nonpathogenic Th17 cells, followed by mass-spectrometry to identify RORγt-associated protein partners and analysis of their functions in RORγt expressing cells. Moreover, due to its nonoverlapping expression patterns with RORγt and obscure functions, RORγ is very interesting to investigate in terms of associated protein complexes and roles in other tissues. RORγt mutations were employed to dissect its roles in cell fate determination and differentiation. We identified a RORγt mutant by random mutagenesis, designated as MUT, which dramatically lost transcriptional activity when expressed in 293T-cells and in vitro polarized Rorc-KO Th17 cells. Mice harboring MUT mutation had normal thymocyte differentiation and development of secondary lymphoid tissues. No lymphomas were detected in MUT mice, unlike in C57BL/6 RORγt-KO mice that develop T-cell lymphomas. Strikingly, T-cells and ILCs differed in proportions and numbers according to tissues. In secondary lymphoid organs, including draining lymph nodes and spleen, no differences were detected in proportions or numbers of CD4 or CD8 T-cells. In large intestine, however, the proportions and numbers of γδ T-cells and Th17 cells were significantly reduced in MUT mice, while those of Th2 cells were markedly increased. Meanwhile, hyper-proliferation of RORγt-negative T helper cells and Treg cells was detected in large intestine of MUT mice. ILCs from large intestine, including ILC3 with high expression of RORγt, were similar to wild type mice in both proportions and numbers. A different phenotype was detected in small intestine, where T-cells and ILCs were markedly reduced. Given the different requirement or expression stage of RORγt for development of T-cells and ILCs, the results probably reflect the effects of RORγt-associated functions on microenvironments. Analyses of immune cells and microbiome at different ages may help to address the mechanism. The functions of MUT RORγt-expressing immune cells will be explored in homeostatic or pathogenic settings with SFB colonization or the EAE model. Moreover, comparison of transcriptomes of wild-type and MUT RORγt expressing cells may reveal essential targets for differentiation and function, which will be validated by transfer of bone marrow or T-cells engineered by CRISPR-mediated gene knockout or retroviral overexpression. Wild-type and MUT RORγt-associated protein complexes will be compared to interrogate potential mechanisms in directing cell differentiation. Detected by mass-spectrometry and biochemical technologies, the majority of RORγt proteins are phosphorylated in naïve thymocytes and in vitro polarized Th17 cells. Mice with respective mutations in RORγt were generated by CRISPR technology. Similar analysis as indicated above will be performed to interrogate roles of RORγt in cell fate determination and differentiation. Citation Format: Hao Xu, Dan Littman, Wendy Huang. Dissection of RORγt functions in lymphoid cell fate determination and differentiation [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B198.

Background /Aims: Wnt are a family of intercellular signaling factors that regulates a wide range of functions during embryonic development including cell proliferation, cell fate determination, differentiation, cell polarity and cell migration. They bind to Frizzled receptors and thereafter proceed via Dishevelled to several intracellular pathways known as the canonical and non-canonical signaling pathways. Activation of the canonical pathway results in an accumulation of cytoplasmic beta-catenin levels. Increased Wnt/beta-catenin signaling is associated with many forms of cancer, including the embryonic neural childhood tumors medulloblastoma and neuroblastoma. The non-canonical Wnt signaling pathways can inhibit the canonical signaling pathway, although the mechanism is still unclear. In this study, we investigate the roles of Prickle1 and Van Gogh-Like 2 (Vangl2), two planar cell polarity (PCP) genes in regulation of the cytoskeleton, Wnt/beta-catenin activity and tumorigenicity in neuroblastoma and medulloblastoma. Methods: mRNA and protein expression of human Prickle1, Vangl2 and beta-catenin were examined in neuroblastoma and medulloblastoma cell lines. siRNA and cDNA transfection were performed together with co-immunoprecipitation and immunocytochemistry. Results and conclusions: We show that Prickle1 and Vangl2 are expressed in neuroblastoma and medulloblastoma cell lines at different levels. Decreased Prickle1 expression correlates with increased levels of Wnt/beta-catenin in both medullo- and neuroblastoma cell lines, demonstrating a downregulation of PCP components and a possible activation of the canonical Wnt signaling pathway. Immunoprecipitation demonstrated that Prickle1 is present in complex with Vangl2. However, the exact mode of interaction between Prickle1 and Vangl2 remains to be eluciated. Microarray analysis revealed that low expressions of Prickle1 or Vangl2 are both associated significantly worse clinical outcome in neuroblastoma. Additionally, siRNA experiments revealed that knockdown of Prickle1 and Vangl2 increased cell proliferation of neuroblastoma cells. Taken together this indicate that the non-canonical Wnt pathway may be an interesting therapeutic target in embryanol neural childhood tumors. Citation Format: Cecilia H. Dyberg, Panagiotis Papachristou, Hugo Lagercrantz, John I Johnsen, Malin Wickström. Vangl2 and Prickle1 expression, interaction and regulation of Wnt/PCP signaling in medulloblastoma and neuroblastoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3795. doi:10.1158/1538-7445.AM2013-3795

1.1 Defining differentiation and cell fate in cancer In the setting of embryonic development, terms such as “cell fate“ and “cellular differentiation“ are relatively easy to grasp, since they refer to a straightforward linear model whereby progenitor cells give rise sequentially to various distinct and identifiable lineages, eventually resulting in a terminally differentiated cell that has until recently been thought to possess very little cellular plasticity or multipotent capacity. The sequences of events that regulate this process are extremely well conserved within a species, and even across species in many instances. In the setting of cancer, cell fate and cellular differentiation are often used descriptively to convey an observed phenotype rather than a defined and well-understood molecular process. How accurate is it to refer to “differentiation“ in cancer when the so-called end-point for this process results in cellular heterogeneity that is antithetical to the regulated and predictable differentiated cells that result from embryonic development? What does it mean to refer to “cell fate“ during the inherently dysregulated series of events involved in oncogenesis, which may not result in a distinct endpoint even across cells within the same tumor? Our review uses these terms in reference to the dynamic processes that constantly shape the function and properties of melanoma cells, which coincidentally utilize many of the same pathways involved in the regulated process of differentiation and determination of cell fate during embryonic development. While the label of being a differentiated cell may imply a terminal nature that might be viewed as less tumorigenic or lethal in the setting of cancer, this concept requires further experimental confirmation. Undoubtedly, these terms will become more refined as our understanding of the molecular events underlying melanoma progression comes into clearer focus. For now, they are the best terms available to describe some of the events and processes that help determine the behavior and response of tumor cells, and we use them with the full knowledge that years from now, advances in our understanding of cancer could render these terms woefully inadequate, or worse yet inaccurate.

Implantation requires reciprocal interaction between blastocysts and a receptive uterus. In mice, one important player in this dialogue involves endocannabinoid signaling via cannabinoid receptor CB1. Anandamide is an endogenous cannabinoid ligand, and its levels are spatiotemporally regulated in the uterus during early pregnancy, showing lower levels in the receptive uterus and at the implantation site. However, the mechanism by which differential uterine anandamide gradients are established under different pregnancy status is not clearly understood. Using multiple approaches, we show here that uterine anandamide levels conducive to implantation are primarily regulated by spatiotemporal expression of Nape-Pld, the gene encoding N-acylphosphatidylethanolamine-hydrolyzing phospholipase D that generates anandamide. The expression is well correlated with its activity and anandamide levels. This study is clinically relevant, since elevated anandamide levels in peripheral circulation are associated with spontaneous pregnancy failure in women.

Wnt family members are best known for their roles in cell fate determination, differentiation, proliferation and apoptosis during embryonic development. Wnt signaling becomes effective during these cellular processes through the proper interaction between its ligands, receptors, effectors and inhibitors. Here we review Wnt signaling in terms of embryonic development to the blastocyst stage implantation with emphasis on endometrial changes that are critical for receptivity in the uterus. The relationship between Wnt signaling and implantation clearly reveals that, Wnt family members are critical for both early embryonic development and changing of the endometrium before implantation. Specific Wnt signaling pathway members are demonstrated to be critical for endometrial events such as decidualization and endometrial gland formation in addition to cyclic changes in the endometrium controlled by reproductive hormones. In conclusion, specific roles of Wnt members and associated factors for both uterine function and embryonic development should be further investigated with respect to the efficiency of human ARTs.

The formation of the neatly layered retina during embryonic development is dictated by a series of complicated transcription factor interactions. Retina-specific expression of these transcription factors is an essential step in establishing retinal progenitor cells (RPCs) from embryonic stem cells. The transcriptional control of gene expression is largely mediated by the combinatorial interactions between cis-regulatory DNA elements and trans-acting transcription factors, which cooperate/interact with each other to form a transcription regulatory network during this developmental process. Such regulatory networks are essential in regulating tissue/cell-specific gene expression, e.g., in cell fate determination and differentiation during embryonic retinal development (Hu et al., 2010; Kumar, 2009; Swaroop et al., 2010). Many genes, which involved in transcriptional networks for the specification of a certain retinal cell lineage, have already been identified and characterized (Corbo et al., 2010; Kim et al., 2008b; Tang et al., 2010). The transcriptional regulatory networks for specific retinal cell lineages, e.g., the photoreceptors (Corbo et al., 2010; Hsiau et al., 2007) and bipolar neurons (Kim et al., 2008b), were established in recent years. However, the transcriptional regulatory network that governs the entire neural retinal development is still elusive. Identifying tissue/cell-specific cis-regulatory elements, trans-acting factor binding sites (TFBSs), and their binding transcription factors (TFs) represent key steps towards understanding tissue/cell-specific gene expression and further successful reconstruction of transcriptional regulatory networks. These steps also present major challenges in both fields of experimental biology and computational biology. Currently, the prevailing method of studying TFBSs and transcriptional regulatory networks is to determine the function of tissue-specific trans-acting factors based on data from genome-wide gene expression profiling and chromatin immunoprecipitation (ChIP). ChIP is often used to investigate protein-DNA interactions in a cell. Coupled with massive parallel sequencing, ChIP-seq is capable of mapping the genome-wide protein-DNA interaction at a finer resolution (Valouev et al., 2008) to identify candidate enhancer sequences (Visel et al., 2009). Thus, a regulatory cascade can be recognized via consequential analysis of the factors involved. Here, we present a new method for the computational analysis of TFBSs and transcriptional regulatory networks utilizing genome-wide sequencing, expression, and enhancer data. In

Ligand-independent activation of the sevenless receptor tyrosine kinase changes the fate of cells in the developing Drosophila eye

BackgroundThe cellular signaling pathway (network) is one of the main topics of organismic investigations. The intracellular interactions between genes in a signaling pathway are considered as the foundation of functional genomics. Thus, what genes and how much they influence each other through transcriptional binding or physical interactions are essential problems. Under the synchronous measures of gene expression via a microarray chip, an amount of dynamic information is embedded and remains to be discovered. Using a systematically dynamic modeling approach, we explore the causal relationship among genes in cellular signaling pathways from the system biology approach.ResultsIn this study, a second-order dynamic model is developed to describe the regulatory mechanism of a target gene from the upstream causality point of view. From the expression profile and dynamic model of a target gene, we can estimate its upstream regulatory function. According to this upstream regulatory function, we would deduce the upstream regulatory genes with their regulatory abilities and activation delays, and then link up a regulatory pathway. Iteratively, these regulatory genes are considered as target genes to trace back their upstream regulatory genes. Then we could construct the regulatory pathway (or network) to the genome wide. In short, we can infer the genetic regulatory pathways from gene-expression profiles quantitatively, which can confirm some doubted paths or seek some unknown paths in a regulatory pathway (network). Finally, the proposed approach is validated by randomly reshuffling the time order of microarray data.ConclusionWe focus our algorithm on the inference of regulatory abilities of the identified causal genes, and how much delay before they regulate the downstream genes. With this information, a regulatory pathway would be built up using microarray data. In the present study, two signaling pathways, i.e. circadian regulatory pathway in Arabidopsis thaliana and metabolic shift pathway from fermentation to respiration in yeast Saccharomyces cerevisiae, are reconstructed using microarray data to evaluate the performance of our proposed method. In the circadian regulatory pathway, we identified mainly the interactions between the biological clock and the photoperiodic genes consistent with the known regulatory mechanisms. We also discovered the now less-known regulations between crytochrome and phytochrome. In the metabolic shift pathway, the casual relationship of enzymatic genes could be detected properly.