Background Nuclear receptors, including estrogen receptor (ER), progesterone receptor (PR)-B, per... more Background Nuclear receptors, including estrogen receptor (ER), progesterone receptor (PR)-B, peroxisome proliferator – activated receptor gamma, and retinoic acid receptor alpha, have been implicated in breast cancer etiology and progression. We investigated the role of dendritic cell – specific transcript (DC-SCRIPT) as coregulator of these nuclear receptors and as a prognostic factor in breast cancer. Methods The effect of DC-SCRIPT on the transcriptional activity of nuclear receptors was assessed by luciferase reporter assays. DC-SCRIPT expression in normal and tumor tissue from breast cancer patients was analyzed by polymerase chain reaction and immunohistochemistry. The prognostic value of tumor DC-SCRIPT mRNA expression was assessed in three independent cohorts of breast cancer patients: a discovery group (n = 47) and a validation group (n = 97) (neither of which had received systemic adjuvant therapy) and in a tamoxifen-treated validation group (n = 68) by using a DC-SCRIPT to porphobilinogen deaminase transcript ratio cutoff of 0.15 determined in the discovery group. Univariate and multivariable Cox proportional hazards model analyses were performed. All statistical tests were two-sided. Results DC-SCRIPT suppressed ER- and PR-mediated transcription in a ligand-dependent fashion, whereas it enhanced the retinoic acid receptor alpha – and peroxisome proliferator – activated receptor gamma – mediated transcription. In breast tissue samples from nine patients , DC-SCRIPT mRNA was expressed at lower levels in the tumor than in the corresponding normal tissue ( P = .010). Patients in the discovery group with high tumor DC-SCRIPT mRNA levels (66%) had a longer disease-free interval than those with a low DC-SCRIPT mRNA level (34%) (hazard ratio [HR] of recurrence for high vs low DC-SCRIPT level = 0.23, 95% confidence interval [CI] = 0.06 to 0.93, P = .039), which was confirmed in the validation group (HR of recurrence = 0.50, 95% CI = 0.26 to 0.95, P = .034). This prognostic value was confined to patients with ER- and/or PR-positive tumors (discovery group: HR of recurrence = 0.16, 95% CI = 0.03 to 0.89, P = .030; validation group: HR of recurrence = 0.42, 95% CI = 0.19 to 0.91, P = .028) and was also observed in the second validation group (HR = 0.46, 95% CI = 0.22 to 0.97, P = .040). DC-SCRIPT was an independent prognostic factor after correction for tumor size, lymph node status, and adjuvant therapy (n = 145; HR = 0.50, 95% CI = 0.29 to 0.85, P = .010).
Histone-modifying enzymes are required for cell identity and lineage commitment, however
little i... more Histone-modifying enzymes are required for cell identity and lineage commitment, however little is known about the regulatory origins of the epigenome during embryonic development. Here we generate a comprehensive set of epigenome reference maps, which we use to determine the extent to which maternal factors shape chromatin state in Xenopus embryos. Using a-amanitin to inhibit zygotic transcription, we find that the majority of H3K4me3- and H3K27me3-enriched regions form a maternally defined epigenetic regulatory space with an underlying logic of hypomethylated islands. This maternal regulatory space extends to a substantial proportion of neurula stage-activated promoters. In contrast, p300 recruitment to distal regulatory regions requires embryonic transcription at most loci. The results show that H3K4me3 and H3K27me3 are part of a regulatory space that exerts an extended maternal control well into post-gastrulation development, and highlight the combinatorial action of maternal and zygotic factors through proximal and distal regulatory sequences.
Polycomb group (PcG) proteins are key regulators in establishing a transcriptional repressive sta... more Polycomb group (PcG) proteins are key regulators in establishing a transcriptional repressive state. Polycomb Repressive Complex 2 (PRC2), one of the two major PcG protein complexes, is essential for proper differentiation and maintenance of cellular identity. Multiple factors are involved in recruiting PRC2 to its genomic targets. In this review, we will discuss the role of DNA sequence, transcription factors, pre-existing histone modifications, and RNA in guiding PRC2 towards specific genomic loci. The DNA sequence itself influences the DNA methylation state, which is an important determinant of PRC2 recruitment. Other histone modifications are also important for PRC2 binding as PRC2 can respond to different cellular states via crosstalk between histone modifications. Additionally, PRC2 might be able to sense the transcriptional status of genes by binding to nascent RNA, which could also guide the complex to chromatin. In this review, we will discuss how all these molecular aspects define a local chromatin state which controls accurate, cell-type-specific epigenetic silencing by PRC2.
The identification of a major class of innate
immune receptors, termed pattern recognition
recept... more The identification of a major class of innate immune receptors, termed pattern recognition receptors (PRRs), has boosted research on innate pathogen recognition. The immune response to a specific pathogen is not restricted to the recognition by one type of PRR or activation of a single cell type, but instead comprises complex collaborations between different receptors, cells and signal mediators. Here we will discuss the crosstalk between PRRs involved in fungal recognition, focusing on the molecular interactions occurring at the plasma membrane.
Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or... more Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or promote tolerance. The DC family is generally comprised of two functionally distinct DC subsets. Conventional dendritic cells (cDC) are the classical antigen presenting cells; plasmacytoid dendritic cells (pDC) are the main producers of type I interferons thereby serving innate immunity. Upon activation DCs are able to present antigen and stimulate T cells. The immune modulatory functions of DCs largely depend on the recognition of soluble cues. Besides pathogen derived cues, recent data indicate that the tissue micro-environment, i.e. of the gut and skin affects cDC function. Many of these micro-environmental factors are ligands for the nuclear receptor (NR) family of transcription regulators known to affect immunity and tolerance. Whether pDC function is also influenced by tissue derived cues, like hormones, vitamins and metabolic products, is largely unknown. Here, we investigated the NR expression profile of murine pDCs and cDCs. We assessed the mRNA levels of 19 NRs of in vitro derived as well as ex vivo isolated DCs from four different lymphoid tissues. We observed that cDCs and pDCs expressed the same repertoire of NRs. Expression levels, however, differed between the two subsets, especially upon maturation of DCs. These data imply that NR ligands do impact pDC function and that their activity might be regulated in a DC-specific manner.
Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or toler... more Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or tolerance depending on diverse factors in the DC environment. Pathogens, but also tissue damage, hormones, and vitamins, affect DC activation and maturation. In particular, glucocorticoids (GCs) are known for their immunosuppressive effect on DCs, creating tolerogenic DCs. GCs activate the type I nuclear receptor (NR) glucocorticoid receptor (GR), followed by induced expression of the transcription factor glucocorticoid-inducible leucine zipper (GILZ). GILZ has been shown to be necessary and sufficient for GC-induced tolerogenic DC generation. Recently, we have identified the DC-specific transcript (DC-SCRIPT) as an NR coregulator, suppressing type I steroid NRs estrogen receptor and progesterone receptor. In this study, we analyzed the effect of DC-SCRIPT on GR activity. We demonstrate that DC-SCRIPT coexists with GR in protein complexes and functions as a corepressor of GR-mediated transcription. Coexpression of DC-SCRIPT and GR is shown in human monocyte-derived DCs, and DC-SCRIPT knockdown enhances GR-dependent upregulation of GILZ mRNA expression in DCs. This demonstrates that DC-SCRIPT serves an important role in regulating GR function in DCs, corepressing GR-dependent upregulation of the tolerance-inducing transcription factor GILZ. These data imply that by controlling GR function and GILZ expression DC-SCRIPT is potentially involved in the balance between tolerance and immunity.
BACKGROUND:
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Recep... more BACKGROUND:
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), play an important role in prostate cancer etiology. We recently found that DC-SCRIPT is a prognostic marker in breast cancer and a unique NR coregulator differentially regulating different classes of NRs. Here we investigated the importance of DC-SCRIPT in prostate cancer. METHODS:
DC-SCRIPT mRNA expression was measured by qPCR. Immunohistochemistry was used to detect DC-SCRIPT protein expression. The functional effects of DC-SCRIPT on the transcriptional activity of AR and VDR were assessed by luciferase reporter assays and qPCR assays on well-known AR and VDR target genes. RESULTS:
DC-SCRIPT mRNA was higher in normal than in corresponding malignant prostate tissue but could not be related to disease stage. DC-SCRIPT protein was found in morphologically normal prostate glands and in infiltrating immune cells. Strikingly, DC-SCRIPT protein expression was absent in malignant prostate epithelial tissue and prostate carcinoma cell lines. DC-SCRIPT protein expression appears to be lost prior to the basal cell marker HMW cytokeratin used in prostate carcinoma diagnostics. In addition, our data demonstrated that DC-SCRIPT repressed transcription mediated by wild-type and mutated AR while enhancing VDR mediated transcription. In addition, transient expression of DC-SCRIPT expression in prostate carcinoma cells strongly repressed cell growth. CONCLUSIONS:
DC-SCRIPT is a key regulator of nuclear receptors AR and VDR that play an opposite role in prostate cancer etiology and loss of DC-SCRIPT may be involved in the onset of prostate cancer.
Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and co... more Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and course of an immune response. Molecular understanding of DC biology is important for the design of DC-based immunotherapies and optimal clinical applications in vaccination settings. Previously, we isolated and characterized the cDNA-encoding dendritic cell-specific transcript (DC-SCRIPT; also known as ZNF366). DC-SCRIPT mRNA expression in the immune system was confined to DCs and was reported to be an early hallmark of DC differentiation. In this study, we demonstrate IL-4 to be the dominant factor for DC-SCRIPT expression in human monocyte-derived DCs. In addition, to our knowledge, we show for the first time endogenous DC-SCRIPT protein expression in human DCs both in vitro and in situ. DC-SCRIPT protein is detected early upon differentiation of monocytes into DCs and is also present in multiple freshly isolated DC subsets. Maturation of DCs with TLR ligands further increased DC-SCRIPT mRNA expression, suggesting a role in DC maturation. Indeed, small interfering RNA-mediated knockdown of DC-SCRIPT affected the cytokine response upon TLR stimulation. These DCs displayed enhanced IL-10 and decreased IL-12 production, compared with wild-type DCs. Silencing of IL-10 in DC-SCRIPT knockdown DCs rescued IL-12 expression, suggesting a primary role for DC-SCRIPT in the regulation of IL-10 production.
The cAMP/PKA signalling pathway and transcription factor cAMP response element-binding protein (C... more The cAMP/PKA signalling pathway and transcription factor cAMP response element-binding protein (CREB) play key roles in long-term memory (LTM) formation. We used two closely related parasitic wasp species, Cotesia glomerata and Cotesia rubecula, which were previously shown to be different in LTM formation, and sequenced at least nine different CREB transcripts in both wasp species. The splicing patterns, functional domains and amino acid sequences were similar to those found in the CREB genes of other organisms. The predicted amino acid sequences of the CREB isoforms were identical in both wasp species. Using real-time quantitative PCR we found that two low abundant CREB transcripts are differentially expressed in the two wasps, whereas the expression levels of high abundant transcripts are similar.
The balance between tolerance and immunity is important for the outcome of an infection or cancer... more The balance between tolerance and immunity is important for the outcome of an infection or cancer, and dendritic cells (DCs) are key regulators of this balance. DC-specific transcript (DC-SCRIPT) is a protein expressed by DCs and has been demonstrated to suppress both TLR-mediated expression of IL-10 and glucocorticoid receptor–mediated transcription of glucocorticoid-induced leucine zipper (GILZ). Because GILZ is known to promote IL-10 production, we investigated whether these two processes are linked. Dual-knockdown and inhibition experiments demonstrated that neither GILZ nor glucocorticoid receptor play a role in TLR-induced IL-10 production after DC-SCRIPT knockdown. The NF-kB pathway is another route involved in IL-10 production after DC activation. Strikingly, inhibition of NF-kB led to a decreased TLR-mediated IL-10 production in DC-SCRIPT knockdown DCs. Moreover, DC-SCRIPT knockdown DCs showed enhanced phosphorylation, acetylation, and IL10 enhancer binding of the NF-kB subunit p65. These data demonstrate that besides nuclear receptor regulation, DC-SCRIPT also modulates activation of NF-kBp65 after TLR activation in human DCs.
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), pl... more Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), play an important role in prostate cancer etiology. We recently found that DC-SCRIPT is a prognostic marker in breast cancer and a unique NR coregulator differentially regulating different classes of NRs. Here we investigated the importance of DC-SCRIPT in prostate cancer. DC-SCRIPT mRNA expression was measured by qPCR. Immunohistochemistry was used to detect DC-SCRIPT protein expression. The functional effects of DC-SCRIPT on the transcriptional activity of AR and VDR were assessed by luciferase reporter assays and qPCR assays on well-known AR and VDR target genes. DC-SCRIPT mRNA was higher in normal than in corresponding malignant prostate tissue but could not be related to disease stage. DC-SCRIPT protein was found in morphologically normal prostate glands and in infiltrating immune cells. Strikingly, DC-SCRIPT protein expression was absent in malignant prostate epithelial tissue and prostate carcinoma cell lines. DC-SCRIPT protein expression appears to be lost prior to the basal cell marker HMW cytokeratin used in prostate carcinoma diagnostics. In addition, our data demonstrated that DC-SCRIPT repressed transcription mediated by wild-type and mutated AR while enhancing VDR mediated transcription. In addition, transient expression of DC-SCRIPT expression in prostate carcinoma cells strongly repressed cell growth. DC-SCRIPT is a key regulator of nuclear receptors AR and VDR that play an opposite role in prostate cancer etiology and loss of DC-SCRIPT may be involved in the onset of prostate cancer.
Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and co... more Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and course of an immune response. Molecular understanding of DC biology is important for the design of DC-based immunotherapies and optimal clinical applications in vaccination settings. Previously, we isolated and characterized the cDNA-encoding dendritic cell-specific transcript (DC-SCRIPT; also known as ZNF366). DC-SCRIPT mRNA expression in the immune system was confined to DCs and was reported to be an early hallmark of DC differentiation. In this study, we demonstrate IL-4 to be the dominant factor for DC-SCRIPT expression in human monocyte-derived DCs. In addition, to our knowledge, we show for the first time endogenous DC-SCRIPT protein expression in human DCs both in vitro and in situ. DC-SCRIPT protein is detected early upon differentiation of monocytes into DCs and is also present in multiple freshly isolated DC subsets. Maturation of DCs with TLR ligands further increased DC-SCRIPT mRNA expression, suggesting a role in DC maturation. Indeed, small interfering RNA-mediated knockdown of DC-SCRIPT affected the cytokine response upon TLR stimulation. These DCs displayed enhanced IL-10 and decreased IL-12 production, compared with wild-type DCs. Silencing of IL-10 in DC-SCRIPT knockdown DCs rescued IL-12 expression, suggesting a primary role for DC-SCRIPT in the regulation of IL-10 production.
Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or toler... more Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or tolerance depending on diverse factors in the DC environment. Pathogens, but also tissue damage, hormones, and vitamins, affect DC activation and maturation. In particular, glucocorticoids (GCs) are known for their immunosuppressive effect on DCs, creating tolerogenic DCs. GCs activate the type I nuclear receptor (NR) glucocorticoid receptor (GR), followed by induced expression of the transcription factor glucocorticoid-inducible leucine zipper (GILZ). GILZ has been shown to be necessary and sufficient for GC-induced tolerogenic DC generation. Recently, we have identified the DC-specific transcript (DC-SCRIPT) as an NR coregulator, suppressing type I steroid NRs estrogen receptor and progesterone receptor. In this study, we analyzed the effect of DC-SCRIPT on GR activity. We demonstrate that DC-SCRIPT coexists with GR in protein complexes and functions as a corepressor of GR-mediated transcription. Coexpression of DC-SCRIPT and GR is shown in human monocyte-derived DCs, and DC-SCRIPT knockdown enhances GR-dependent upregulation of GILZ mRNA expression in DCs. This demonstrates that DC-SCRIPT serves an important role in regulating GR function in DCs, corepressing GR-dependent upregulation of the tolerance-inducing transcription factor GILZ. These data imply that by controlling GR function and GILZ expression DC-SCRIPT is potentially involved in the balance between tolerance and immunity.
The balance between tolerance and immunity is important for the outcome of an infection or cancer... more The balance between tolerance and immunity is important for the outcome of an infection or cancer, and dendritic cells (DCs) are key regulators of this balance. DC-specific transcript (DC-SCRIPT) is a protein expressed by DCs and has been demonstrated to suppress both TLR-mediated expression of IL-10 and glucocorticoid receptor-mediated transcription of glucocorticoid-induced leucine zipper (GILZ). Because GILZ is known to promote IL-10 production, we investigated whether these two processes are linked. Dual-knockdown and inhibition experiments demonstrated that neither GILZ nor glucocorticoid receptor play a role in TLR-induced IL-10 production after DC-SCRIPT knockdown. The NF-κB pathway is another route involved in IL-10 production after DC activation. Strikingly, inhibition of NF-κB led to a decreased TLR-mediated IL-10 production in DC-SCRIPT knockdown DCs. Moreover, DC-SCRIPT knockdown DCs showed enhanced phosphorylation, acetylation, and IL10 enhancer binding of the NF-κB subunit p65. These data demonstrate that besides nuclear receptor regulation, DC-SCRIPT also modulates activation of NF-κBp65 after TLR activation in human DCs.
Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or... more Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or promote tolerance. The DC family is generally comprised of two functionally distinct DC subsets. Conventional dendritic cells (cDC) are the classical antigen presenting cells; plasmacytoid dendritic cells (pDC) are the main producers of type I interferons thereby serving innate immunity. Upon activation DCs are able to present antigen and stimulate T cells. The immune modulatory functions of DCs largely depend on the recognition of soluble cues. Besides pathogen derived cues, recent data indicate that the tissue micro-environment, i.e. of the gut and skin affects cDC function. Many of these micro-environmental factors are ligands for the nuclear receptor (NR) family of transcription regulators known to affect immunity and tolerance. Whether pDC function is also influenced by tissue derived cues, like hormones, vitamins and metabolic products, is largely unknown. Here, we investigated the NR expression profile of murine pDCs and cDCs. We assessed the mRNA levels of 19 NRs of in vitro derived as well as ex vivo isolated DCs from four different lymphoid tissues. We observed that cDCs and pDCs expressed the same repertoire of NRs. Expression levels, however, differed between the two subsets, especially upon maturation of DCs. These data imply that NR ligands do impact pDC function and that their activity might be regulated in a DC-specific manner.
Background Nuclear receptors, including estrogen receptor (ER), progesterone receptor (PR)-B, per... more Background Nuclear receptors, including estrogen receptor (ER), progesterone receptor (PR)-B, peroxisome proliferator – activated receptor gamma, and retinoic acid receptor alpha, have been implicated in breast cancer etiology and progression. We investigated the role of dendritic cell – specific transcript (DC-SCRIPT) as coregulator of these nuclear receptors and as a prognostic factor in breast cancer. Methods The effect of DC-SCRIPT on the transcriptional activity of nuclear receptors was assessed by luciferase reporter assays. DC-SCRIPT expression in normal and tumor tissue from breast cancer patients was analyzed by polymerase chain reaction and immunohistochemistry. The prognostic value of tumor DC-SCRIPT mRNA expression was assessed in three independent cohorts of breast cancer patients: a discovery group (n = 47) and a validation group (n = 97) (neither of which had received systemic adjuvant therapy) and in a tamoxifen-treated validation group (n = 68) by using a DC-SCRIPT to porphobilinogen deaminase transcript ratio cutoff of 0.15 determined in the discovery group. Univariate and multivariable Cox proportional hazards model analyses were performed. All statistical tests were two-sided. Results DC-SCRIPT suppressed ER- and PR-mediated transcription in a ligand-dependent fashion, whereas it enhanced the retinoic acid receptor alpha – and peroxisome proliferator – activated receptor gamma – mediated transcription. In breast tissue samples from nine patients , DC-SCRIPT mRNA was expressed at lower levels in the tumor than in the corresponding normal tissue ( P = .010). Patients in the discovery group with high tumor DC-SCRIPT mRNA levels (66%) had a longer disease-free interval than those with a low DC-SCRIPT mRNA level (34%) (hazard ratio [HR] of recurrence for high vs low DC-SCRIPT level = 0.23, 95% confidence interval [CI] = 0.06 to 0.93, P = .039), which was confirmed in the validation group (HR of recurrence = 0.50, 95% CI = 0.26 to 0.95, P = .034). This prognostic value was confined to patients with ER- and/or PR-positive tumors (discovery group: HR of recurrence = 0.16, 95% CI = 0.03 to 0.89, P = .030; validation group: HR of recurrence = 0.42, 95% CI = 0.19 to 0.91, P = .028) and was also observed in the second validation group (HR = 0.46, 95% CI = 0.22 to 0.97, P = .040). DC-SCRIPT was an independent prognostic factor after correction for tumor size, lymph node status, and adjuvant therapy (n = 145; HR = 0.50, 95% CI = 0.29 to 0.85, P = .010).
Histone-modifying enzymes are required for cell identity and lineage commitment, however
little i... more Histone-modifying enzymes are required for cell identity and lineage commitment, however little is known about the regulatory origins of the epigenome during embryonic development. Here we generate a comprehensive set of epigenome reference maps, which we use to determine the extent to which maternal factors shape chromatin state in Xenopus embryos. Using a-amanitin to inhibit zygotic transcription, we find that the majority of H3K4me3- and H3K27me3-enriched regions form a maternally defined epigenetic regulatory space with an underlying logic of hypomethylated islands. This maternal regulatory space extends to a substantial proportion of neurula stage-activated promoters. In contrast, p300 recruitment to distal regulatory regions requires embryonic transcription at most loci. The results show that H3K4me3 and H3K27me3 are part of a regulatory space that exerts an extended maternal control well into post-gastrulation development, and highlight the combinatorial action of maternal and zygotic factors through proximal and distal regulatory sequences.
Polycomb group (PcG) proteins are key regulators in establishing a transcriptional repressive sta... more Polycomb group (PcG) proteins are key regulators in establishing a transcriptional repressive state. Polycomb Repressive Complex 2 (PRC2), one of the two major PcG protein complexes, is essential for proper differentiation and maintenance of cellular identity. Multiple factors are involved in recruiting PRC2 to its genomic targets. In this review, we will discuss the role of DNA sequence, transcription factors, pre-existing histone modifications, and RNA in guiding PRC2 towards specific genomic loci. The DNA sequence itself influences the DNA methylation state, which is an important determinant of PRC2 recruitment. Other histone modifications are also important for PRC2 binding as PRC2 can respond to different cellular states via crosstalk between histone modifications. Additionally, PRC2 might be able to sense the transcriptional status of genes by binding to nascent RNA, which could also guide the complex to chromatin. In this review, we will discuss how all these molecular aspects define a local chromatin state which controls accurate, cell-type-specific epigenetic silencing by PRC2.
The identification of a major class of innate
immune receptors, termed pattern recognition
recept... more The identification of a major class of innate immune receptors, termed pattern recognition receptors (PRRs), has boosted research on innate pathogen recognition. The immune response to a specific pathogen is not restricted to the recognition by one type of PRR or activation of a single cell type, but instead comprises complex collaborations between different receptors, cells and signal mediators. Here we will discuss the crosstalk between PRRs involved in fungal recognition, focusing on the molecular interactions occurring at the plasma membrane.
Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or... more Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or promote tolerance. The DC family is generally comprised of two functionally distinct DC subsets. Conventional dendritic cells (cDC) are the classical antigen presenting cells; plasmacytoid dendritic cells (pDC) are the main producers of type I interferons thereby serving innate immunity. Upon activation DCs are able to present antigen and stimulate T cells. The immune modulatory functions of DCs largely depend on the recognition of soluble cues. Besides pathogen derived cues, recent data indicate that the tissue micro-environment, i.e. of the gut and skin affects cDC function. Many of these micro-environmental factors are ligands for the nuclear receptor (NR) family of transcription regulators known to affect immunity and tolerance. Whether pDC function is also influenced by tissue derived cues, like hormones, vitamins and metabolic products, is largely unknown. Here, we investigated the NR expression profile of murine pDCs and cDCs. We assessed the mRNA levels of 19 NRs of in vitro derived as well as ex vivo isolated DCs from four different lymphoid tissues. We observed that cDCs and pDCs expressed the same repertoire of NRs. Expression levels, however, differed between the two subsets, especially upon maturation of DCs. These data imply that NR ligands do impact pDC function and that their activity might be regulated in a DC-specific manner.
Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or toler... more Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or tolerance depending on diverse factors in the DC environment. Pathogens, but also tissue damage, hormones, and vitamins, affect DC activation and maturation. In particular, glucocorticoids (GCs) are known for their immunosuppressive effect on DCs, creating tolerogenic DCs. GCs activate the type I nuclear receptor (NR) glucocorticoid receptor (GR), followed by induced expression of the transcription factor glucocorticoid-inducible leucine zipper (GILZ). GILZ has been shown to be necessary and sufficient for GC-induced tolerogenic DC generation. Recently, we have identified the DC-specific transcript (DC-SCRIPT) as an NR coregulator, suppressing type I steroid NRs estrogen receptor and progesterone receptor. In this study, we analyzed the effect of DC-SCRIPT on GR activity. We demonstrate that DC-SCRIPT coexists with GR in protein complexes and functions as a corepressor of GR-mediated transcription. Coexpression of DC-SCRIPT and GR is shown in human monocyte-derived DCs, and DC-SCRIPT knockdown enhances GR-dependent upregulation of GILZ mRNA expression in DCs. This demonstrates that DC-SCRIPT serves an important role in regulating GR function in DCs, corepressing GR-dependent upregulation of the tolerance-inducing transcription factor GILZ. These data imply that by controlling GR function and GILZ expression DC-SCRIPT is potentially involved in the balance between tolerance and immunity.
BACKGROUND:
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Recep... more BACKGROUND:
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), play an important role in prostate cancer etiology. We recently found that DC-SCRIPT is a prognostic marker in breast cancer and a unique NR coregulator differentially regulating different classes of NRs. Here we investigated the importance of DC-SCRIPT in prostate cancer. METHODS:
DC-SCRIPT mRNA expression was measured by qPCR. Immunohistochemistry was used to detect DC-SCRIPT protein expression. The functional effects of DC-SCRIPT on the transcriptional activity of AR and VDR were assessed by luciferase reporter assays and qPCR assays on well-known AR and VDR target genes. RESULTS:
DC-SCRIPT mRNA was higher in normal than in corresponding malignant prostate tissue but could not be related to disease stage. DC-SCRIPT protein was found in morphologically normal prostate glands and in infiltrating immune cells. Strikingly, DC-SCRIPT protein expression was absent in malignant prostate epithelial tissue and prostate carcinoma cell lines. DC-SCRIPT protein expression appears to be lost prior to the basal cell marker HMW cytokeratin used in prostate carcinoma diagnostics. In addition, our data demonstrated that DC-SCRIPT repressed transcription mediated by wild-type and mutated AR while enhancing VDR mediated transcription. In addition, transient expression of DC-SCRIPT expression in prostate carcinoma cells strongly repressed cell growth. CONCLUSIONS:
DC-SCRIPT is a key regulator of nuclear receptors AR and VDR that play an opposite role in prostate cancer etiology and loss of DC-SCRIPT may be involved in the onset of prostate cancer.
Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and co... more Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and course of an immune response. Molecular understanding of DC biology is important for the design of DC-based immunotherapies and optimal clinical applications in vaccination settings. Previously, we isolated and characterized the cDNA-encoding dendritic cell-specific transcript (DC-SCRIPT; also known as ZNF366). DC-SCRIPT mRNA expression in the immune system was confined to DCs and was reported to be an early hallmark of DC differentiation. In this study, we demonstrate IL-4 to be the dominant factor for DC-SCRIPT expression in human monocyte-derived DCs. In addition, to our knowledge, we show for the first time endogenous DC-SCRIPT protein expression in human DCs both in vitro and in situ. DC-SCRIPT protein is detected early upon differentiation of monocytes into DCs and is also present in multiple freshly isolated DC subsets. Maturation of DCs with TLR ligands further increased DC-SCRIPT mRNA expression, suggesting a role in DC maturation. Indeed, small interfering RNA-mediated knockdown of DC-SCRIPT affected the cytokine response upon TLR stimulation. These DCs displayed enhanced IL-10 and decreased IL-12 production, compared with wild-type DCs. Silencing of IL-10 in DC-SCRIPT knockdown DCs rescued IL-12 expression, suggesting a primary role for DC-SCRIPT in the regulation of IL-10 production.
The cAMP/PKA signalling pathway and transcription factor cAMP response element-binding protein (C... more The cAMP/PKA signalling pathway and transcription factor cAMP response element-binding protein (CREB) play key roles in long-term memory (LTM) formation. We used two closely related parasitic wasp species, Cotesia glomerata and Cotesia rubecula, which were previously shown to be different in LTM formation, and sequenced at least nine different CREB transcripts in both wasp species. The splicing patterns, functional domains and amino acid sequences were similar to those found in the CREB genes of other organisms. The predicted amino acid sequences of the CREB isoforms were identical in both wasp species. Using real-time quantitative PCR we found that two low abundant CREB transcripts are differentially expressed in the two wasps, whereas the expression levels of high abundant transcripts are similar.
The balance between tolerance and immunity is important for the outcome of an infection or cancer... more The balance between tolerance and immunity is important for the outcome of an infection or cancer, and dendritic cells (DCs) are key regulators of this balance. DC-specific transcript (DC-SCRIPT) is a protein expressed by DCs and has been demonstrated to suppress both TLR-mediated expression of IL-10 and glucocorticoid receptor–mediated transcription of glucocorticoid-induced leucine zipper (GILZ). Because GILZ is known to promote IL-10 production, we investigated whether these two processes are linked. Dual-knockdown and inhibition experiments demonstrated that neither GILZ nor glucocorticoid receptor play a role in TLR-induced IL-10 production after DC-SCRIPT knockdown. The NF-kB pathway is another route involved in IL-10 production after DC activation. Strikingly, inhibition of NF-kB led to a decreased TLR-mediated IL-10 production in DC-SCRIPT knockdown DCs. Moreover, DC-SCRIPT knockdown DCs showed enhanced phosphorylation, acetylation, and IL10 enhancer binding of the NF-kB subunit p65. These data demonstrate that besides nuclear receptor regulation, DC-SCRIPT also modulates activation of NF-kBp65 after TLR activation in human DCs.
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), pl... more Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), play an important role in prostate cancer etiology. We recently found that DC-SCRIPT is a prognostic marker in breast cancer and a unique NR coregulator differentially regulating different classes of NRs. Here we investigated the importance of DC-SCRIPT in prostate cancer. DC-SCRIPT mRNA expression was measured by qPCR. Immunohistochemistry was used to detect DC-SCRIPT protein expression. The functional effects of DC-SCRIPT on the transcriptional activity of AR and VDR were assessed by luciferase reporter assays and qPCR assays on well-known AR and VDR target genes. DC-SCRIPT mRNA was higher in normal than in corresponding malignant prostate tissue but could not be related to disease stage. DC-SCRIPT protein was found in morphologically normal prostate glands and in infiltrating immune cells. Strikingly, DC-SCRIPT protein expression was absent in malignant prostate epithelial tissue and prostate carcinoma cell lines. DC-SCRIPT protein expression appears to be lost prior to the basal cell marker HMW cytokeratin used in prostate carcinoma diagnostics. In addition, our data demonstrated that DC-SCRIPT repressed transcription mediated by wild-type and mutated AR while enhancing VDR mediated transcription. In addition, transient expression of DC-SCRIPT expression in prostate carcinoma cells strongly repressed cell growth. DC-SCRIPT is a key regulator of nuclear receptors AR and VDR that play an opposite role in prostate cancer etiology and loss of DC-SCRIPT may be involved in the onset of prostate cancer.
Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and co... more Dendritic cells (DCs) are the professional APCs of the immune system that dictate the type and course of an immune response. Molecular understanding of DC biology is important for the design of DC-based immunotherapies and optimal clinical applications in vaccination settings. Previously, we isolated and characterized the cDNA-encoding dendritic cell-specific transcript (DC-SCRIPT; also known as ZNF366). DC-SCRIPT mRNA expression in the immune system was confined to DCs and was reported to be an early hallmark of DC differentiation. In this study, we demonstrate IL-4 to be the dominant factor for DC-SCRIPT expression in human monocyte-derived DCs. In addition, to our knowledge, we show for the first time endogenous DC-SCRIPT protein expression in human DCs both in vitro and in situ. DC-SCRIPT protein is detected early upon differentiation of monocytes into DCs and is also present in multiple freshly isolated DC subsets. Maturation of DCs with TLR ligands further increased DC-SCRIPT mRNA expression, suggesting a role in DC maturation. Indeed, small interfering RNA-mediated knockdown of DC-SCRIPT affected the cytokine response upon TLR stimulation. These DCs displayed enhanced IL-10 and decreased IL-12 production, compared with wild-type DCs. Silencing of IL-10 in DC-SCRIPT knockdown DCs rescued IL-12 expression, suggesting a primary role for DC-SCRIPT in the regulation of IL-10 production.
Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or toler... more Dendritic cells (DCs) play a central role in the immune system; they can induce immunity or tolerance depending on diverse factors in the DC environment. Pathogens, but also tissue damage, hormones, and vitamins, affect DC activation and maturation. In particular, glucocorticoids (GCs) are known for their immunosuppressive effect on DCs, creating tolerogenic DCs. GCs activate the type I nuclear receptor (NR) glucocorticoid receptor (GR), followed by induced expression of the transcription factor glucocorticoid-inducible leucine zipper (GILZ). GILZ has been shown to be necessary and sufficient for GC-induced tolerogenic DC generation. Recently, we have identified the DC-specific transcript (DC-SCRIPT) as an NR coregulator, suppressing type I steroid NRs estrogen receptor and progesterone receptor. In this study, we analyzed the effect of DC-SCRIPT on GR activity. We demonstrate that DC-SCRIPT coexists with GR in protein complexes and functions as a corepressor of GR-mediated transcription. Coexpression of DC-SCRIPT and GR is shown in human monocyte-derived DCs, and DC-SCRIPT knockdown enhances GR-dependent upregulation of GILZ mRNA expression in DCs. This demonstrates that DC-SCRIPT serves an important role in regulating GR function in DCs, corepressing GR-dependent upregulation of the tolerance-inducing transcription factor GILZ. These data imply that by controlling GR function and GILZ expression DC-SCRIPT is potentially involved in the balance between tolerance and immunity.
The balance between tolerance and immunity is important for the outcome of an infection or cancer... more The balance between tolerance and immunity is important for the outcome of an infection or cancer, and dendritic cells (DCs) are key regulators of this balance. DC-specific transcript (DC-SCRIPT) is a protein expressed by DCs and has been demonstrated to suppress both TLR-mediated expression of IL-10 and glucocorticoid receptor-mediated transcription of glucocorticoid-induced leucine zipper (GILZ). Because GILZ is known to promote IL-10 production, we investigated whether these two processes are linked. Dual-knockdown and inhibition experiments demonstrated that neither GILZ nor glucocorticoid receptor play a role in TLR-induced IL-10 production after DC-SCRIPT knockdown. The NF-κB pathway is another route involved in IL-10 production after DC activation. Strikingly, inhibition of NF-κB led to a decreased TLR-mediated IL-10 production in DC-SCRIPT knockdown DCs. Moreover, DC-SCRIPT knockdown DCs showed enhanced phosphorylation, acetylation, and IL10 enhancer binding of the NF-κB subunit p65. These data demonstrate that besides nuclear receptor regulation, DC-SCRIPT also modulates activation of NF-κBp65 after TLR activation in human DCs.
Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or... more Dendritic cells (DC) play a central role in the immune system. They can either induce immunity or promote tolerance. The DC family is generally comprised of two functionally distinct DC subsets. Conventional dendritic cells (cDC) are the classical antigen presenting cells; plasmacytoid dendritic cells (pDC) are the main producers of type I interferons thereby serving innate immunity. Upon activation DCs are able to present antigen and stimulate T cells. The immune modulatory functions of DCs largely depend on the recognition of soluble cues. Besides pathogen derived cues, recent data indicate that the tissue micro-environment, i.e. of the gut and skin affects cDC function. Many of these micro-environmental factors are ligands for the nuclear receptor (NR) family of transcription regulators known to affect immunity and tolerance. Whether pDC function is also influenced by tissue derived cues, like hormones, vitamins and metabolic products, is largely unknown. Here, we investigated the NR expression profile of murine pDCs and cDCs. We assessed the mRNA levels of 19 NRs of in vitro derived as well as ex vivo isolated DCs from four different lymphoid tissues. We observed that cDCs and pDCs expressed the same repertoire of NRs. Expression levels, however, differed between the two subsets, especially upon maturation of DCs. These data imply that NR ligands do impact pDC function and that their activity might be regulated in a DC-specific manner.
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activated receptor gamma, and retinoic acid receptor alpha, have been implicated in breast cancer etiology
and progression. We investigated the role of dendritic cell – specific transcript (DC-SCRIPT) as coregulator of
these nuclear receptors and as a prognostic factor in breast cancer.
Methods The effect of DC-SCRIPT on the transcriptional activity of nuclear receptors was assessed by luciferase
reporter assays. DC-SCRIPT expression in normal and tumor tissue from breast cancer patients was analyzed
by polymerase chain reaction and immunohistochemistry. The prognostic value of tumor DC-SCRIPT
mRNA expression was assessed in three independent cohorts of breast cancer patients: a discovery group
(n = 47) and a validation group (n = 97) (neither of which had received systemic adjuvant therapy) and in
a tamoxifen-treated validation group (n = 68) by using a DC-SCRIPT to porphobilinogen deaminase transcript
ratio cutoff of 0.15 determined in the discovery group. Univariate and multivariable Cox proportional
hazards model analyses were performed. All statistical tests were two-sided.
Results DC-SCRIPT suppressed ER- and PR-mediated transcription in a ligand-dependent fashion, whereas it enhanced
the retinoic acid receptor alpha – and peroxisome proliferator – activated receptor gamma – mediated transcription.
In breast tissue samples from nine patients , DC-SCRIPT mRNA was expressed at lower levels in
the tumor than in the corresponding normal tissue ( P = .010). Patients in the discovery group with high
tumor DC-SCRIPT mRNA levels (66%) had a longer disease-free interval than those with a low DC-SCRIPT
mRNA level (34%) (hazard ratio [HR] of recurrence for high vs low DC-SCRIPT level = 0.23, 95% confidence
interval [CI] = 0.06 to 0.93, P = .039), which was confirmed in the validation group (HR of recurrence = 0.50,
95% CI = 0.26 to 0.95, P = .034). This prognostic value was confined to patients with ER- and/or PR-positive
tumors (discovery group: HR of recurrence = 0.16, 95% CI = 0.03 to 0.89, P = .030; validation group: HR of
recurrence = 0.42, 95% CI = 0.19 to 0.91, P = .028) and was also observed in the second validation group
(HR = 0.46, 95% CI = 0.22 to 0.97, P = .040). DC-SCRIPT was an independent prognostic factor after correction
for tumor size, lymph node status, and adjuvant therapy (n = 145; HR = 0.50, 95% CI = 0.29 to 0.85, P = .010).
little is known about the regulatory origins of the epigenome during embryonic development.
Here we generate a comprehensive set of epigenome reference maps, which we use to
determine the extent to which maternal factors shape chromatin state in Xenopus embryos.
Using a-amanitin to inhibit zygotic transcription, we find that the majority of H3K4me3- and
H3K27me3-enriched regions form a maternally defined epigenetic regulatory space with an
underlying logic of hypomethylated islands. This maternal regulatory space extends to a
substantial proportion of neurula stage-activated promoters. In contrast, p300 recruitment to
distal regulatory regions requires embryonic transcription at most loci. The results show that
H3K4me3 and H3K27me3 are part of a regulatory space that exerts an extended maternal
control well into post-gastrulation development, and highlight the combinatorial action of
maternal and zygotic factors through proximal and distal regulatory sequences.
Repressive Complex 2 (PRC2), one of the two major PcG protein complexes, is essential for proper
differentiation and maintenance of cellular identity. Multiple factors are involved in recruiting PRC2
to its genomic targets. In this review, we will discuss the role of DNA sequence, transcription factors,
pre-existing histone modifications, and RNA in guiding PRC2 towards specific genomic loci. The DNA
sequence itself influences the DNA methylation state, which is an important determinant of PRC2 recruitment.
Other histone modifications are also important for PRC2 binding as PRC2 can respond to different
cellular states via crosstalk between histone modifications. Additionally, PRC2 might be able to sense
the transcriptional status of genes by binding to nascent RNA, which could also guide the complex to
chromatin. In this review, we will discuss how all these molecular aspects define a local chromatin state
which controls accurate, cell-type-specific epigenetic silencing by PRC2.
immune receptors, termed pattern recognition
receptors (PRRs), has boosted research on innate
pathogen recognition. The immune response to a
specific pathogen is not restricted to the recognition
by one type of PRR or activation of a single
cell type, but instead comprises complex collaborations
between different receptors, cells and
signal mediators. Here we will discuss the crosstalk
between PRRs involved in fungal recognition,
focusing on the molecular interactions occurring
at the plasma membrane.
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), play an important role in prostate cancer etiology. We recently found that DC-SCRIPT is a prognostic marker in breast cancer and a unique NR coregulator differentially regulating different classes of NRs. Here we investigated the importance of DC-SCRIPT in prostate cancer.
METHODS:
DC-SCRIPT mRNA expression was measured by qPCR. Immunohistochemistry was used to detect DC-SCRIPT protein expression. The functional effects of DC-SCRIPT on the transcriptional activity of AR and VDR were assessed by luciferase reporter assays and qPCR assays on well-known AR and VDR target genes.
RESULTS:
DC-SCRIPT mRNA was higher in normal than in corresponding malignant prostate tissue but could not be related to disease stage. DC-SCRIPT protein was found in morphologically normal prostate glands and in infiltrating immune cells. Strikingly, DC-SCRIPT protein expression was absent in malignant prostate epithelial tissue and prostate carcinoma cell lines. DC-SCRIPT protein expression appears to be lost prior to the basal cell marker HMW cytokeratin used in prostate carcinoma diagnostics. In addition, our data demonstrated that DC-SCRIPT repressed transcription mediated by wild-type and mutated AR while enhancing VDR mediated transcription. In addition, transient expression of DC-SCRIPT expression in prostate carcinoma cells strongly repressed cell growth.
CONCLUSIONS:
DC-SCRIPT is a key regulator of nuclear receptors AR and VDR that play an opposite role in prostate cancer etiology and loss of DC-SCRIPT may be involved in the onset of prostate cancer.
key regulators of this balance. DC-specific transcript (DC-SCRIPT) is a protein expressed by DCs and has been demonstrated to
suppress both TLR-mediated expression of IL-10 and glucocorticoid receptor–mediated transcription of glucocorticoid-induced
leucine zipper (GILZ). Because GILZ is known to promote IL-10 production, we investigated whether these two processes are
linked. Dual-knockdown and inhibition experiments demonstrated that neither GILZ nor glucocorticoid receptor play a role in
TLR-induced IL-10 production after DC-SCRIPT knockdown. The NF-kB pathway is another route involved in IL-10 production
after DC activation. Strikingly, inhibition of NF-kB led to a decreased TLR-mediated IL-10 production in DC-SCRIPT knockdown
DCs. Moreover, DC-SCRIPT knockdown DCs showed enhanced phosphorylation, acetylation, and IL10 enhancer binding of
the NF-kB subunit p65. These data demonstrate that besides nuclear receptor regulation, DC-SCRIPT also modulates activation
of NF-kBp65 after TLR activation in human DCs.
activated receptor gamma, and retinoic acid receptor alpha, have been implicated in breast cancer etiology
and progression. We investigated the role of dendritic cell – specific transcript (DC-SCRIPT) as coregulator of
these nuclear receptors and as a prognostic factor in breast cancer.
Methods The effect of DC-SCRIPT on the transcriptional activity of nuclear receptors was assessed by luciferase
reporter assays. DC-SCRIPT expression in normal and tumor tissue from breast cancer patients was analyzed
by polymerase chain reaction and immunohistochemistry. The prognostic value of tumor DC-SCRIPT
mRNA expression was assessed in three independent cohorts of breast cancer patients: a discovery group
(n = 47) and a validation group (n = 97) (neither of which had received systemic adjuvant therapy) and in
a tamoxifen-treated validation group (n = 68) by using a DC-SCRIPT to porphobilinogen deaminase transcript
ratio cutoff of 0.15 determined in the discovery group. Univariate and multivariable Cox proportional
hazards model analyses were performed. All statistical tests were two-sided.
Results DC-SCRIPT suppressed ER- and PR-mediated transcription in a ligand-dependent fashion, whereas it enhanced
the retinoic acid receptor alpha – and peroxisome proliferator – activated receptor gamma – mediated transcription.
In breast tissue samples from nine patients , DC-SCRIPT mRNA was expressed at lower levels in
the tumor than in the corresponding normal tissue ( P = .010). Patients in the discovery group with high
tumor DC-SCRIPT mRNA levels (66%) had a longer disease-free interval than those with a low DC-SCRIPT
mRNA level (34%) (hazard ratio [HR] of recurrence for high vs low DC-SCRIPT level = 0.23, 95% confidence
interval [CI] = 0.06 to 0.93, P = .039), which was confirmed in the validation group (HR of recurrence = 0.50,
95% CI = 0.26 to 0.95, P = .034). This prognostic value was confined to patients with ER- and/or PR-positive
tumors (discovery group: HR of recurrence = 0.16, 95% CI = 0.03 to 0.89, P = .030; validation group: HR of
recurrence = 0.42, 95% CI = 0.19 to 0.91, P = .028) and was also observed in the second validation group
(HR = 0.46, 95% CI = 0.22 to 0.97, P = .040). DC-SCRIPT was an independent prognostic factor after correction
for tumor size, lymph node status, and adjuvant therapy (n = 145; HR = 0.50, 95% CI = 0.29 to 0.85, P = .010).
little is known about the regulatory origins of the epigenome during embryonic development.
Here we generate a comprehensive set of epigenome reference maps, which we use to
determine the extent to which maternal factors shape chromatin state in Xenopus embryos.
Using a-amanitin to inhibit zygotic transcription, we find that the majority of H3K4me3- and
H3K27me3-enriched regions form a maternally defined epigenetic regulatory space with an
underlying logic of hypomethylated islands. This maternal regulatory space extends to a
substantial proportion of neurula stage-activated promoters. In contrast, p300 recruitment to
distal regulatory regions requires embryonic transcription at most loci. The results show that
H3K4me3 and H3K27me3 are part of a regulatory space that exerts an extended maternal
control well into post-gastrulation development, and highlight the combinatorial action of
maternal and zygotic factors through proximal and distal regulatory sequences.
Repressive Complex 2 (PRC2), one of the two major PcG protein complexes, is essential for proper
differentiation and maintenance of cellular identity. Multiple factors are involved in recruiting PRC2
to its genomic targets. In this review, we will discuss the role of DNA sequence, transcription factors,
pre-existing histone modifications, and RNA in guiding PRC2 towards specific genomic loci. The DNA
sequence itself influences the DNA methylation state, which is an important determinant of PRC2 recruitment.
Other histone modifications are also important for PRC2 binding as PRC2 can respond to different
cellular states via crosstalk between histone modifications. Additionally, PRC2 might be able to sense
the transcriptional status of genes by binding to nascent RNA, which could also guide the complex to
chromatin. In this review, we will discuss how all these molecular aspects define a local chromatin state
which controls accurate, cell-type-specific epigenetic silencing by PRC2.
immune receptors, termed pattern recognition
receptors (PRRs), has boosted research on innate
pathogen recognition. The immune response to a
specific pathogen is not restricted to the recognition
by one type of PRR or activation of a single
cell type, but instead comprises complex collaborations
between different receptors, cells and
signal mediators. Here we will discuss the crosstalk
between PRRs involved in fungal recognition,
focusing on the molecular interactions occurring
at the plasma membrane.
Nuclear receptors (NR), including the Androgen Receptor (AR) and the Vitamin D Receptor (VDR), play an important role in prostate cancer etiology. We recently found that DC-SCRIPT is a prognostic marker in breast cancer and a unique NR coregulator differentially regulating different classes of NRs. Here we investigated the importance of DC-SCRIPT in prostate cancer.
METHODS:
DC-SCRIPT mRNA expression was measured by qPCR. Immunohistochemistry was used to detect DC-SCRIPT protein expression. The functional effects of DC-SCRIPT on the transcriptional activity of AR and VDR were assessed by luciferase reporter assays and qPCR assays on well-known AR and VDR target genes.
RESULTS:
DC-SCRIPT mRNA was higher in normal than in corresponding malignant prostate tissue but could not be related to disease stage. DC-SCRIPT protein was found in morphologically normal prostate glands and in infiltrating immune cells. Strikingly, DC-SCRIPT protein expression was absent in malignant prostate epithelial tissue and prostate carcinoma cell lines. DC-SCRIPT protein expression appears to be lost prior to the basal cell marker HMW cytokeratin used in prostate carcinoma diagnostics. In addition, our data demonstrated that DC-SCRIPT repressed transcription mediated by wild-type and mutated AR while enhancing VDR mediated transcription. In addition, transient expression of DC-SCRIPT expression in prostate carcinoma cells strongly repressed cell growth.
CONCLUSIONS:
DC-SCRIPT is a key regulator of nuclear receptors AR and VDR that play an opposite role in prostate cancer etiology and loss of DC-SCRIPT may be involved in the onset of prostate cancer.
key regulators of this balance. DC-specific transcript (DC-SCRIPT) is a protein expressed by DCs and has been demonstrated to
suppress both TLR-mediated expression of IL-10 and glucocorticoid receptor–mediated transcription of glucocorticoid-induced
leucine zipper (GILZ). Because GILZ is known to promote IL-10 production, we investigated whether these two processes are
linked. Dual-knockdown and inhibition experiments demonstrated that neither GILZ nor glucocorticoid receptor play a role in
TLR-induced IL-10 production after DC-SCRIPT knockdown. The NF-kB pathway is another route involved in IL-10 production
after DC activation. Strikingly, inhibition of NF-kB led to a decreased TLR-mediated IL-10 production in DC-SCRIPT knockdown
DCs. Moreover, DC-SCRIPT knockdown DCs showed enhanced phosphorylation, acetylation, and IL10 enhancer binding of
the NF-kB subunit p65. These data demonstrate that besides nuclear receptor regulation, DC-SCRIPT also modulates activation
of NF-kBp65 after TLR activation in human DCs.