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Gary S. Stein - One of the best experts on this subject based on the ideXlab platform.
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microrna 466 inhibits tumor growth and bone metastasis in prostate cancer by direct regulation of osteogenic Transcription Factor RUNX2
Cell Death and Disease, 2018Co-Authors: Melissa Colden, Gary S. Stein, Sharanjot Saini, Priya V Dahiya, Varahram Shahryari, Soichiro Yamamura, Yuichiro Tanaka, Rajvir Dahiya, Shahana MajidAbstract:MicroRNAs (miRNAs) have emerged as key players in cancer progression and metastatic initiation yet their importance in regulating prostate cancer (PCa) metastasis to bone has begun to be appreciated. We employed multimodal strategy based on in-house PCa clinical samples, publicly available TCGA cohorts, a panel of cell lines, in silico analyses, and a series of in vitro and in vivo assays to investigate the role of miR-466 in PCa. Expression analyses revealed that miR-466 is under-expressed in PCa compared to normal tissues. Reconstitution of miR-466 in metastatic PCa cell lines impaired their oncogenic functions such as cell proliferation, migration/invasion and induced cell cycle arrest, and apoptosis compared to control miRNA. Conversely, attenuation of miR-466 in normal prostate cells induced tumorigenic characteristics. miR-466 suppressed PCa growth and metastasis through direct targeting of bone-related Transcription Factor RUNX2. Overexpression of miR-466 caused a marked downregulation of integrated network of RUNX2 target genes such as osteopontin, osteocalcin, ANGPTs, MMP11 including Fyn, pAkt, FAK and vimentin that are known to be involved in migration, invasion, angiogenesis, EMT and metastasis. Xenograft models indicate that miR-466 inhibits primary orthotopic tumor growth and spontaneous metastasis to bone. Receiver operating curve and Kaplan–Meier analyses show that miR-466 expression can discriminate between malignant and normal prostate tissues; and can predict biochemical relapse. In conclusion, our data strongly suggests miR-466-mediated attenuation of RUNX2 as a novel therapeutic approach to regulate PCa growth, particularly metastasis to bone. This study is the first report documenting the anti-bone metastatic role and clinical significance of miR-466 in prostate cancer.
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mitotic inheritance of mrna facilitates translational activation of the osteogenic lineage commitment Factor RUNX2 in progeny of osteoblastic cells
Journal of Cellular Physiology, 2016Co-Authors: Nelson Varela, Marcelo Antonelli, Martin Montecino, Gary S. Stein, Hugo Sepulveda, Carlos Lizama, Roman Thaler, Alejandra Aranguiz, Ricardo D Moreno, Andre J Van WijnenAbstract:Epigenetic mechanisms mediate the acquisition of specialized cellular phenotypes during tissue development, maintenance and repair. When phenotype-committed cells transit through mitosis, chromosomal condensation counteracts epigenetic activation of gene expression. Subsequent post-mitotic re-activation of Transcription depends on epigenetic DNA and histone modifications, as well as other architecturally bound proteins that “bookmark” the genome. Osteogenic lineage commitment, differentiation and progenitor proliferation require the bone-related runt-related Transcription Factor RUNX2. Here, we characterized a non-genomic mRNA mediated mechanism by which osteoblast precursors retain their phenotype during self-renewal. We show that osteoblasts produce maximal levels of RUNX2 mRNA, but not protein, prior to mitotic cell division. RUNX2 mRNA partitions symmetrically between daughter cells in a non-chromosomal tubulin-containing compartment. Subsequently, Transcription-independent de novo synthesis of RUNX2 protein in early G1 phase results in increased functional interactions of RUNX2 with a representative osteoblast-specific target gene (osteocalcin/BGLAP2) in chromatin. Somatic transmission of RUNX2 mRNAs in osteoblasts and osteosarcoma cells represents a versatile mechanism for translational rather than Transcriptional induction of this principal gene regulator to maintain osteoblast phenotype identity after mitosis. J. Cell. Physiol. 231: 1001–1014, 2016. © 2015 Wiley Periodicals, Inc.
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epigenetic control of the bone master RUNX2 gene during osteoblast lineage commitment by the histone demethylase jarid1b kdm5b
Journal of Biological Chemistry, 2015Co-Authors: Jane B Lian, Andre J Van Wijnen, Gary S. Stein, Janet L. Stein, Adriana Rojas, Rodrigo Aguilar, Berta Henriquez, Brigitte Van Zundert, Miguel L AllendeAbstract:Abstract Transcription Factor RUNX2 controls bone development and osteoblast differentiation by regulating expression of a significant number of bone-related target genes. Here, we report that Transcriptional activation and repression of the RUNX2 gene via its osteoblast-specific P1 promoter (encoding mRNA for the RUNX2/p57 isoform) is accompanied by selective deposition and elimination of histone marks during differentiation of mesenchymal cells to the osteogenic and myoblastic lineages. These epigenetic profiles are mediated by key components of the Trithorax/COMPASS-like and Polycomb group complexes together with histone arginine methylases like PRMT5 and lysine demethylases like JARID1B/KDM5B. Importantly, knockdown of the H3K4me2/3 demethylase JARID1B, but not of the demethylases UTX and NO66, prevents repression of the RUNX2 P1 promoter during myogenic differentiation of mesenchymal cells. The epigenetically forced expression of RUNX2/p57 and osteocalcin, a classical bone-related target gene, under myoblastic-differentiation is accompanied by enrichment of the H3K4me3 and H3K27ac marks at the RUNX2 P1 promoter region. Our results identify JARID1B as a key component of a potent epigenetic switch that controls mesenchymal cell fate into myogenic and osteogenic lineages.
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Oncogenic cooperation between PI3K/Akt signaling and Transcription Factor RUNX2 promotes the invasive properties of metastatic breast cancer cells
Journal of Cellular Physiology, 2013Co-Authors: Sandhya Pande, Sayyed K Zaidi, Jane B Lian, Andre J Van Wijnen, Janet L. Stein, Gillian Browne, Srivatsan Padmanabhan, Gary S. SteinAbstract:The serine/threonine kinase Akt/PKB promotes cancer cell growth and invasion through several downstream targets. Identification of novel substrates may provide new avenues for therapeutic intervention. Our study shows that Akt phosphorylates the cancer-related Transcription Factor RUNX2 resulting in stimulated DNA binding of the purified recombinant protein in vitro. Pharmacological inhibition of the PI3K/Akt pathway in breast cancer cells reduces DNA-binding activity of RUNX2 with concomitant reduction in the expression of metastasis-related RUNX2 target genes. Akt phosphorylates RUNX2 at three critical residues within the runt DNA-binding domain to enhance its in vivo genomic interactions with a target gene promoter, MMP13. Mutation of these three phosphorylation sites reduces RUNX2 DNA-binding activity. However, Akt signaling does not appear to interefere with CBFβ-RUNX2 interactions. Consequently, expression of multiple metastasis-related genes is decreased and RUNX2-mediated cell invasion is supressed. Thus, our work identifies RUNX2 as a novel and important downstream mediator of the PI3K/Akt pathway that is linked to metastatic properties of breast cancer cells. J. Cell. Physiol. 228: 1784–1792, 2013. © 2013 Wiley Periodicals, Inc.
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the cancer related Transcription Factor RUNX2 modulates cell proliferation in human osteosarcoma cell lines
Journal of Cellular Physiology, 2013Co-Authors: Claudia M J Lucero, Oscar A Vega, Mariana Osorio, Julio C Tapia, Marcelo Antonelli, Andre J Van Wijnen, Gary S. Stein, Mario GalindoAbstract:Osteosarcoma is the most common bone tumor in children and adolescents (Young and Miller, 1975). The highest incidence of osteosarcoma is in the second decade of life, which suggests a relationship between bone growth and tumor development (Fraumeni, 1967; Cotterill et al., 2004). One of the critical steps for normal skeletal development and bone formation is the proliferative expansion of mesenchymal cells, osteoprogenitors, and immature osteoblasts. Cell growth and differentiation of normal osteoprogenitors and pre-osteoblasts is tightly regulated by RUNX2, which favors a quiescent state (Pratap et al., 2003; Galindo et al., 2005). The growth suppressive potential of RUNX2 is controlled by modulation of its protein levels during the cell cycle (Galindo et al., 2005, 2007). Cell cycle dependent changes of RUNX2 levels occur with respect to G1 progression at a cell cycle stage when normal osteoblasts monitor extra-cellular cues for competency to initiate cell cycle progression beyond the G1/S phase transition. Accordingly, transient RUNX2 overexpression in synchronized cells delays cell cycle entry into S phase and significantly decreases cell proliferation in the MC3T3 pre-osteoblasts, RUNX2 null calvarian osteoprogenitors, C2C12 pluripotent mesenchymal, and IMR-90 fibroblasts cell lines (Pratap et al., 2003; Galindo et al., 2005; Young et al., 2007a; Teplyuk et al., 2008, 2009a). The function of RUNX2 as a negative regulator of cell proliferation is also reflected by linkage of RUNX2 deficiency to cell immortalization and tumorigenesis (Kilbey et al., 2007; Zaidi et al., 2007a). Apart from the growth suppressive potential that is evident during late G1 in osteoblasts (Pratap et al., 2003; Galindo et al., 2005), RUNX2 may have mitogenic potential in early G1 (Teplyuk et al., 2008). Several studies indicate that RUNX2-dependent control of proliferation is cell type-specific. RUNX2 inhibits proliferation of osteoprogenitors and committed osteoblasts (Pratap et al., 2003; Galindo et al., 2005), but it may have distinct biological roles in chondrocytes (Galindo et al., 2005; Hinoi et al., 2006; Komori, 2008) and endothelial cells (Inman and Shore, 2003; Qiao et al., 2006). While immature osteoblasts from mice with RUNX2 null mutations show accelerated proliferative potential, chondrocyte proliferation seems to be decreased in RUNX2 null mice (Pratap et al., 2003; Yoshida et al., 2004), suggesting that RUNX2 would also have opposites roles in different bone cell types. Moreover, ectopic expression of RUNX2 in aortic endothelial cells increases cell proliferation (Sun et al., 2004), whereas RUNX2 depletion inhibits cell proliferation in human marrow endothelial cells (Qiao et al., 2006). These findings support the concept that RUNX2 protein can function as either a bona fide tumor suppressor or a classical oncoprotein depending on the cellular context (Blyth et al., 2005). Current evidence indicates that RUNX2 expression is a key pathological Factor in osteosarcoma (Martin et al., 2011) by controlling a number of cancer-related genes (van der Deen et al., 2012). Moreover, osteosarcoma development may be associated with RUNX2 overexpression and defects in osteogenic differentiation (Wagner et al., 2011). Over-expression of RUNX2 in transgenic mice within the osteoblast lineage inhibits osteoblast maturation, increases bone resorption, and causes osteopenia with multiple fractures (Liu et al., 2001; Geoffroy et al., 2002). RUNX2 is also clearly detected in clinical osteosarcoma samples (Andela et al., 2005; Lu et al., 2008; Sadikovic et al., 2009; Won et al., 2009; Kurek et al., 2010). Analysis of genomic DNA from osteosarcoma patients with amplication of the 6p12#x02013;p21 chromosomal interval, which spans the RUNX2 locus, increases the RUNX2 gene copy number and aberrantly elevates RUNX2 expression (Lau et al., 2004; Lu et al., 2008; Sadikovic et al., 2009). Increased expression of RUNX2 in osteosarcoma biopsies has been associated to increased tumorigenicity, tumor progression, metastases, lower survival, and poor prognosis (Won et al., 2009; Kurek et al., 2010; Sadikovic et al., 2010). Interestingly, osteosarcoma cell culture models may exhibit a similar variability of RUNX2 gene expression, because RUNX2 is expressed at different levels in a number of human osteosarcoma cell lines (Thomas et al., 2004; Lu et al., 2008; Luo et al., 2008; Kurek et al., 2010; Shapovalov et al., 2010). A subset of patient-derived osteosarcoma cell lines exhibit high levels of RUNX2, whereas others show decreased RUNX2 expression in accordance with the findings of Thomas and colleagues who suggested that RUNX2 protein levels are negatively regulated in some types of osteosarcoma (Thomas et al., 2004; Nathan et al., 2009; Pereira et al., 2009; San Martin et al., 2009; Won et al., 2009; Kurek et al., 2010; Sadikovic et al., 2010). Recently, we presented data indicating that cell cycle control of RUNX2, which is readily observed in osteoblasts, is deregulated in osteosarcoma cells (Galindo et al., 2005; San Martin et al., 2009). RUNX2 is constitutively expressed throughout the cell cycle in at least two osteosarcoma (human SaOS and rat ROS) cell lines (Young et al., 2007b; San Martin et al., 2009). Hence, the Transcriptional and post-Transcriptional mechanisms that mediate cell cycle control of RUNX2 gene expression in osteoblasts could be compromised in osteosarcoma cells. The latter may occur in conjunction with abrogation of other molecular mechanisms cells that mediate normal osteoblast proliferation and that may bypass the growth suppressive properties of RUNX2 in bone cancer cells (Nathan et al., 2009). In this article, we systematically examined human osteosarcoma cell lines with respect to RUNX2 gene expression and cell cycle regulation to understand the biological functions of RUNX2 in osteosarcoma cell proliferation. Our main finding is that forced expression of RUNX2 suppresses growth in all cell lines, indicating that stimulation of RUNX2 beyond its preestablished levels in osteosarcoma cells remains capable of triggering an anti-proliferative response. We propose that osteosarcoma cells in which RUNX2 is present must balance prooncogenic functions of RUNX2 with the requirement to maintain RUNX2 at levels that avoid tumor suppression.
Jane B Lian - One of the best experts on this subject based on the ideXlab platform.
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epigenetic control of the bone master RUNX2 gene during osteoblast lineage commitment by the histone demethylase jarid1b kdm5b
Journal of Biological Chemistry, 2015Co-Authors: Jane B Lian, Andre J Van Wijnen, Gary S. Stein, Janet L. Stein, Adriana Rojas, Rodrigo Aguilar, Berta Henriquez, Brigitte Van Zundert, Miguel L AllendeAbstract:Abstract Transcription Factor RUNX2 controls bone development and osteoblast differentiation by regulating expression of a significant number of bone-related target genes. Here, we report that Transcriptional activation and repression of the RUNX2 gene via its osteoblast-specific P1 promoter (encoding mRNA for the RUNX2/p57 isoform) is accompanied by selective deposition and elimination of histone marks during differentiation of mesenchymal cells to the osteogenic and myoblastic lineages. These epigenetic profiles are mediated by key components of the Trithorax/COMPASS-like and Polycomb group complexes together with histone arginine methylases like PRMT5 and lysine demethylases like JARID1B/KDM5B. Importantly, knockdown of the H3K4me2/3 demethylase JARID1B, but not of the demethylases UTX and NO66, prevents repression of the RUNX2 P1 promoter during myogenic differentiation of mesenchymal cells. The epigenetically forced expression of RUNX2/p57 and osteocalcin, a classical bone-related target gene, under myoblastic-differentiation is accompanied by enrichment of the H3K4me3 and H3K27ac marks at the RUNX2 P1 promoter region. Our results identify JARID1B as a key component of a potent epigenetic switch that controls mesenchymal cell fate into myogenic and osteogenic lineages.
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oncogenic cooperation between pi3k akt signaling and Transcription Factor RUNX2 promotes the invasive properties of metastatic breast cancer cells
Journal of Cellular Physiology, 2013Co-Authors: Sandhya Pande, Sayyed K Zaidi, Jane B Lian, Andre J Van Wijnen, Gillian Browne, Srivatsan PadmanabhanAbstract:The serine/threonine kinase Akt/PKB promotes cancer cell growth and invasion through several downstream targets. Identification of novel substrates may provide new avenues for therapeutic intervention. Our study shows that Akt phosphorylates the cancer-related Transcription Factor RUNX2 resulting in stimulated DNA binding of the purified recombinant protein in vitro. Pharmacological inhibition of the PI3K/Akt pathway in breast cancer cells reduces DNA-binding activity of RUNX2 with concomitant reduction in the expression of metastasis-related RUNX2 target genes. Akt phosphorylates RUNX2 at three critical residues within the runt DNA-binding domain to enhance its in vivo genomic interactions with a target gene promoter, MMP13. Mutation of these three phosphorylation sites reduces RUNX2 DNA-binding activity. However, Akt signaling does not appear to interefere with CBFβ-RUNX2 interactions. Consequently, expression of multiple metastasis-related genes is decreased and RUNX2-mediated cell invasion is supressed. Thus, our work identifies RUNX2 as a novel and important downstream mediator of the PI3K/Akt pathway that is linked to metastatic properties of breast cancer cells. J. Cell. Physiol. 228: 1784–1792, 2013. © 2013 Wiley Periodicals, Inc.
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Oncogenic cooperation between PI3K/Akt signaling and Transcription Factor RUNX2 promotes the invasive properties of metastatic breast cancer cells
Journal of Cellular Physiology, 2013Co-Authors: Sandhya Pande, Sayyed K Zaidi, Jane B Lian, Andre J Van Wijnen, Janet L. Stein, Gillian Browne, Srivatsan Padmanabhan, Gary S. SteinAbstract:The serine/threonine kinase Akt/PKB promotes cancer cell growth and invasion through several downstream targets. Identification of novel substrates may provide new avenues for therapeutic intervention. Our study shows that Akt phosphorylates the cancer-related Transcription Factor RUNX2 resulting in stimulated DNA binding of the purified recombinant protein in vitro. Pharmacological inhibition of the PI3K/Akt pathway in breast cancer cells reduces DNA-binding activity of RUNX2 with concomitant reduction in the expression of metastasis-related RUNX2 target genes. Akt phosphorylates RUNX2 at three critical residues within the runt DNA-binding domain to enhance its in vivo genomic interactions with a target gene promoter, MMP13. Mutation of these three phosphorylation sites reduces RUNX2 DNA-binding activity. However, Akt signaling does not appear to interefere with CBFβ-RUNX2 interactions. Consequently, expression of multiple metastasis-related genes is decreased and RUNX2-mediated cell invasion is supressed. Thus, our work identifies RUNX2 as a novel and important downstream mediator of the PI3K/Akt pathway that is linked to metastatic properties of breast cancer cells. J. Cell. Physiol. 228: 1784–1792, 2013. © 2013 Wiley Periodicals, Inc.
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genomic promoter occupancy of runt related Transcription Factor RUNX2 in osteosarcoma cells identifies genes involved in cell adhesion and motility
Journal of Biological Chemistry, 2012Co-Authors: Margaretha Van Der Deen, Jacqueline Akech, Sneha Gupta, Martin Montecino, Jane B Lian, Mario Galindo, Daniel W Young, David S Lapointe, Janet L. Stein, Gary S. SteinAbstract:Abstract Runt-related Transcription Factors (RUNX1, RUNX2, and RUNX3) are key lineage-specific regulators of progenitor cell growth and differentiation but also function pathologically as cancer genes that contribute to tumorigenesis. RUNX2 attenuates growth and stimulates maturation of osteoblasts during bone formation but is also robustly expressed in a subset of osteosarcomas, as well as in metastatic breast and prostate tumors. To assess the biological function of RUNX2 in osteosarcoma cells, we examined human genomic promoter interactions for RUNX2 using chromatin immunoprecipitation (ChIP)-microarray analysis in SAOS-2 cells. Promoter binding of both RUNX2 and RNA polymerase II was compared with gene expression profiles of cells in which RUNX2 was depleted by RNA interference. Many RUNX2-bound loci (1550 of 2339 total) exhibit promoter occupancy by RNA polymerase II and contain the RUNX consensus motif 5′-((T/A/C)G(T/A/C)GG(T/G). Gene ontology analysis indicates that RUNX2 controls components of multiple signaling pathways (e.g. WNT, TGFβ, TNFα, and interleukins), as well as genes linked to cell motility and adhesion (e.g. the focal adhesion-related genes FAK/PTK2 and TLN1). Our results reveal that siRNA depletion of RUNX2, PTK2, or TLN1 diminishes motility of U2OS osteosarcoma cells. Thus, RUNX2 binding to diverse gene loci may support the biological properties of osteosarcoma cells.
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a program of micrornas controls osteogenic lineage progression by targeting Transcription Factor RUNX2
Proceedings of the National Academy of Sciences of the United States of America, 2011Co-Authors: Ying Zhang, Jane B Lian, Andre J Van Wijnen, Janet L. Stein, Carlo M. Croce, Gary S. SteinAbstract:Lineage progression in osteoblasts and chondrocytes is stringently controlled by the cell-fate–determining Transcription Factor RUNX2. In this study, we directly addressed whether microRNAs (miRNAs) can control the osteogenic activity of RUNX2 and affect osteoblast maturation. A panel of 11 RUNX2-targeting miRNAs (miR-23a, miR-30c, miR-34c, miR-133a, miR-135a, miR-137, miR-204, miR-205, miR-217, miR-218, and miR-338) is expressed in a lineage-related pattern in mesenchymal cell types. During both osteogenic and chondrogenic differentiation, these miRNAs, in general, are inversely expressed relative to RUNX2. Based on 3′UTR luciferase reporter, immunoblot, and mRNA stability assays, each miRNA directly attenuates RUNX2 protein accumulation. RUNX2-targeting miRNAs differentially inhibit RUNX2 protein expression in osteoblasts and chondrocytes and display different efficacies. Thus, cellular context contributes to miRNA-mediated regulation of RUNX2. All RUNX2-targeting miRNAs (except miR-218) significantly impede osteoblast differentiation, and their effects can be reversed by the corresponding anti-miRNAs. These findings demonstrate that osteoblastogenesis is limited by an elaborate network of functionally tested miRNAs that directly target the osteogenic master regulator RUNX2.
Andre J Van Wijnen - One of the best experts on this subject based on the ideXlab platform.
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mitotic inheritance of mrna facilitates translational activation of the osteogenic lineage commitment Factor RUNX2 in progeny of osteoblastic cells
Journal of Cellular Physiology, 2016Co-Authors: Nelson Varela, Marcelo Antonelli, Martin Montecino, Gary S. Stein, Hugo Sepulveda, Carlos Lizama, Roman Thaler, Alejandra Aranguiz, Ricardo D Moreno, Andre J Van WijnenAbstract:Epigenetic mechanisms mediate the acquisition of specialized cellular phenotypes during tissue development, maintenance and repair. When phenotype-committed cells transit through mitosis, chromosomal condensation counteracts epigenetic activation of gene expression. Subsequent post-mitotic re-activation of Transcription depends on epigenetic DNA and histone modifications, as well as other architecturally bound proteins that “bookmark” the genome. Osteogenic lineage commitment, differentiation and progenitor proliferation require the bone-related runt-related Transcription Factor RUNX2. Here, we characterized a non-genomic mRNA mediated mechanism by which osteoblast precursors retain their phenotype during self-renewal. We show that osteoblasts produce maximal levels of RUNX2 mRNA, but not protein, prior to mitotic cell division. RUNX2 mRNA partitions symmetrically between daughter cells in a non-chromosomal tubulin-containing compartment. Subsequently, Transcription-independent de novo synthesis of RUNX2 protein in early G1 phase results in increased functional interactions of RUNX2 with a representative osteoblast-specific target gene (osteocalcin/BGLAP2) in chromatin. Somatic transmission of RUNX2 mRNAs in osteoblasts and osteosarcoma cells represents a versatile mechanism for translational rather than Transcriptional induction of this principal gene regulator to maintain osteoblast phenotype identity after mitosis. J. Cell. Physiol. 231: 1001–1014, 2016. © 2015 Wiley Periodicals, Inc.
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epigenetic control of the bone master RUNX2 gene during osteoblast lineage commitment by the histone demethylase jarid1b kdm5b
Journal of Biological Chemistry, 2015Co-Authors: Jane B Lian, Andre J Van Wijnen, Gary S. Stein, Janet L. Stein, Adriana Rojas, Rodrigo Aguilar, Berta Henriquez, Brigitte Van Zundert, Miguel L AllendeAbstract:Abstract Transcription Factor RUNX2 controls bone development and osteoblast differentiation by regulating expression of a significant number of bone-related target genes. Here, we report that Transcriptional activation and repression of the RUNX2 gene via its osteoblast-specific P1 promoter (encoding mRNA for the RUNX2/p57 isoform) is accompanied by selective deposition and elimination of histone marks during differentiation of mesenchymal cells to the osteogenic and myoblastic lineages. These epigenetic profiles are mediated by key components of the Trithorax/COMPASS-like and Polycomb group complexes together with histone arginine methylases like PRMT5 and lysine demethylases like JARID1B/KDM5B. Importantly, knockdown of the H3K4me2/3 demethylase JARID1B, but not of the demethylases UTX and NO66, prevents repression of the RUNX2 P1 promoter during myogenic differentiation of mesenchymal cells. The epigenetically forced expression of RUNX2/p57 and osteocalcin, a classical bone-related target gene, under myoblastic-differentiation is accompanied by enrichment of the H3K4me3 and H3K27ac marks at the RUNX2 P1 promoter region. Our results identify JARID1B as a key component of a potent epigenetic switch that controls mesenchymal cell fate into myogenic and osteogenic lineages.
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RUNX3 facilitates growth of Ewing sarcoma cells.
Journal of Cellular Physiology, 2014Co-Authors: Krista L. Bledsoe, Meghan E. Mcgee-lawrence, Emily T. Camilleri, Scott M. Riester, Andre M. Oliveira, Andre J Van Wijnen, Xiaoke Wang, Jennifer J WestendorfAbstract:Ewing sarcoma is an aggressive pediatric small round cell tumor that predominantly occurs in bone. Approximately 85% of Ewing sarcomas harbor the EWS/FLI fusion protein, which arises from a chromosomal translocation, t(11:22)(q24:q12). EWS/FLI interacts with numerous lineage-essential Transcription Factors to maintain mesenchymal progenitors in an undifferentiated state. We previously showed that EWS/FLI binds the osteogenic Transcription Factor RUNX2 and prevents osteoblast differentiation. In this study, we investigated the role of another Runt-domain protein, RUNX3, in Ewing sarcoma. RUNX3 participates in mesenchymal-derived bone formation and is a context dependent tumor suppressor and oncogene. RUNX3 was detected in all Ewing sarcoma cells examined, whereas RUNX2 was detected in only 73% of specimens. Like RUNX2, RUNX3 binds to EWS/FLI via its Runt domain. EWS/FLI prevented RUNX3 from activating the Transcription of a RUNX-responsive reporter, p6OSE2. Stable suppression of RUNX3 expression in the Ewing sarcoma cell line A673 delayed colony growth in anchorage independent soft agar assays and reversed expression of EWS/FLI-responsive genes. These results demonstrate an important role for RUNX3 in Ewing sarcoma.
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oncogenic cooperation between pi3k akt signaling and Transcription Factor RUNX2 promotes the invasive properties of metastatic breast cancer cells
Journal of Cellular Physiology, 2013Co-Authors: Sandhya Pande, Sayyed K Zaidi, Jane B Lian, Andre J Van Wijnen, Gillian Browne, Srivatsan PadmanabhanAbstract:The serine/threonine kinase Akt/PKB promotes cancer cell growth and invasion through several downstream targets. Identification of novel substrates may provide new avenues for therapeutic intervention. Our study shows that Akt phosphorylates the cancer-related Transcription Factor RUNX2 resulting in stimulated DNA binding of the purified recombinant protein in vitro. Pharmacological inhibition of the PI3K/Akt pathway in breast cancer cells reduces DNA-binding activity of RUNX2 with concomitant reduction in the expression of metastasis-related RUNX2 target genes. Akt phosphorylates RUNX2 at three critical residues within the runt DNA-binding domain to enhance its in vivo genomic interactions with a target gene promoter, MMP13. Mutation of these three phosphorylation sites reduces RUNX2 DNA-binding activity. However, Akt signaling does not appear to interefere with CBFβ-RUNX2 interactions. Consequently, expression of multiple metastasis-related genes is decreased and RUNX2-mediated cell invasion is supressed. Thus, our work identifies RUNX2 as a novel and important downstream mediator of the PI3K/Akt pathway that is linked to metastatic properties of breast cancer cells. J. Cell. Physiol. 228: 1784–1792, 2013. © 2013 Wiley Periodicals, Inc.
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Oncogenic cooperation between PI3K/Akt signaling and Transcription Factor RUNX2 promotes the invasive properties of metastatic breast cancer cells
Journal of Cellular Physiology, 2013Co-Authors: Sandhya Pande, Sayyed K Zaidi, Jane B Lian, Andre J Van Wijnen, Janet L. Stein, Gillian Browne, Srivatsan Padmanabhan, Gary S. SteinAbstract:The serine/threonine kinase Akt/PKB promotes cancer cell growth and invasion through several downstream targets. Identification of novel substrates may provide new avenues for therapeutic intervention. Our study shows that Akt phosphorylates the cancer-related Transcription Factor RUNX2 resulting in stimulated DNA binding of the purified recombinant protein in vitro. Pharmacological inhibition of the PI3K/Akt pathway in breast cancer cells reduces DNA-binding activity of RUNX2 with concomitant reduction in the expression of metastasis-related RUNX2 target genes. Akt phosphorylates RUNX2 at three critical residues within the runt DNA-binding domain to enhance its in vivo genomic interactions with a target gene promoter, MMP13. Mutation of these three phosphorylation sites reduces RUNX2 DNA-binding activity. However, Akt signaling does not appear to interefere with CBFβ-RUNX2 interactions. Consequently, expression of multiple metastasis-related genes is decreased and RUNX2-mediated cell invasion is supressed. Thus, our work identifies RUNX2 as a novel and important downstream mediator of the PI3K/Akt pathway that is linked to metastatic properties of breast cancer cells. J. Cell. Physiol. 228: 1784–1792, 2013. © 2013 Wiley Periodicals, Inc.
Yabing Chen - One of the best experts on this subject based on the ideXlab platform.
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smooth muscle cell specific RUNX2 deficiency inhibits vascular calcification
Circulation Research, 2012Co-Authors: Chang Hyun Byon, Amjad Javed, Jianfeng Chen, Kaiyu Yuan, Jack M Heath, Peter G Anderson, Kui Zhang, Yabing ChenAbstract:Rationale:Vascular calcification is a hallmark of atherosclerosis, a major cause of morbidity and mortality in the United States. We have previously reported that the osteogenic Transcription Factor RUNX2 is an essential and sufficient regulator of calcification of vascular smooth muscle cells (VSMC) in vitro. Objective:To determine the contribution of osteogenic differentiation of VSMC to the pathogenesis of vascular calcification and the function of VSMC-derived RUNX2 in regulating calcification in vivo. Methods and Results:SMC-specific RUNX2-deficient mice, generated by breeding SM22α-Cre mice with the RUNX2 exon 8 floxed mice, exhibited normal aortic gross anatomy and expression levels of SMC-specific marker genes. RUNX2 deficiency did not affect basal SMC markers, but inhibited oxidative stress-reduced expression of SMC markers. High-fat-diet-induced vascular calcification in vivo was markedly inhibited in the RUNX2-deficient mice in comparison with their control littermates. RUNX2 deficiency inhibit...
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RUNX2 upregulated receptor activator of nuclear Factor κb ligand in calcifying smooth muscle cells promotes migration and osteoclastic differentiation of macrophages
Arteriosclerosis Thrombosis and Vascular Biology, 2011Co-Authors: Chang Hyun Byon, Jianfeng Chen, Kaiyu Yuan, Jack M Heath, Peter G Anderson, Yin Tintut, Linda L Demer, Deli Wang, Yabing ChenAbstract:Objective—Clinical and experimental studies demonstrate the important roles of vascular smooth muscle cells (VSMC) in the pathogenesis of atherosclerosis. We have previously determined that the osteogenic Transcription Factor RUNX2 is essential for VSMC calcification. The present study characterized RUNX2-regulated signals and their potential roles in vascular calcification. Methods and Results—In vivo studies with atherogenic apolipoprotein E−/− mice demonstrated that increased oxidative stress was associated with upregulation of RUNX2 and receptor activator of nuclear Factor κB ligand (RANKL), which colocalized in the calcified atherosclerotic lesions and were juxtaposed to infiltrated macrophages and osteoclast-like cells that are positively stained for an osteoclast marker, tartrate-resistant acid phosphatase. Mechanistic studies using RNA interference, a luciferase reporter system, chromatin immunoprecipitation, and electrophoretic mobility shift assays indicated that RUNX2 regulated the expression o...
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oxidative stress induces vascular calcification through modulation of the osteogenic Transcription Factor RUNX2 by akt signaling
Journal of Biological Chemistry, 2008Co-Authors: Chang Hyun Byon, Amjad Javed, John C Kappes, Thomas L Clemens, Victor M Darleyusmar, Jay M Mcdonald, Yabing ChenAbstract:Oxidative stress plays a critical role in the pathogenesis of atherosclerosis including the formation of lipid laden macrophages and the development of inflammation. However, oxidative stress-induced molecular signaling that regulates the development of vascular calcification has not been investigated in depth. Osteogenic differentiation of vascular smooth muscle cells (VSMC) is critical in the development of calcification in atherosclerotic lesions. An important contributor to oxidative stress in atherosclerotic lesions is the formation of hydrogen peroxide from diverse sources in vascular cells. In this study we defined molecular signaling that is operative in the H2O2-induced VSMC calcification. We found that H2O2 promotes a phenotypic switch of VSMC from contractile to osteogenic phenotype. This response was associated with an increased expression and transactivity of RUNX2, a key Transcription Factor for osteogenic differentiation. The essential role of RUNX2 in oxidative stress-induced VSMC calcification was further confirmed by RUNX2 depletion and overexpression. Inhibition of RUNX2 using short hairpin RNA blocked VSMC calcification, and adenovirus-mediated overexpression of RUNX2 alone induced VSMC calcification. Inhibition of H2O2-activated AKT signaling blocked VSMC calcification and RUNX2 induction concurrently. This blockage did not cause VSMC apoptosis. Taken together, our data demonstrate a critical role for AKT-mediated induction of RUNX2 in oxidative stress-induced VSMC calcification.
Janet L. Stein - One of the best experts on this subject based on the ideXlab platform.
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epigenetic control of the bone master RUNX2 gene during osteoblast lineage commitment by the histone demethylase jarid1b kdm5b
Journal of Biological Chemistry, 2015Co-Authors: Jane B Lian, Andre J Van Wijnen, Gary S. Stein, Janet L. Stein, Adriana Rojas, Rodrigo Aguilar, Berta Henriquez, Brigitte Van Zundert, Miguel L AllendeAbstract:Abstract Transcription Factor RUNX2 controls bone development and osteoblast differentiation by regulating expression of a significant number of bone-related target genes. Here, we report that Transcriptional activation and repression of the RUNX2 gene via its osteoblast-specific P1 promoter (encoding mRNA for the RUNX2/p57 isoform) is accompanied by selective deposition and elimination of histone marks during differentiation of mesenchymal cells to the osteogenic and myoblastic lineages. These epigenetic profiles are mediated by key components of the Trithorax/COMPASS-like and Polycomb group complexes together with histone arginine methylases like PRMT5 and lysine demethylases like JARID1B/KDM5B. Importantly, knockdown of the H3K4me2/3 demethylase JARID1B, but not of the demethylases UTX and NO66, prevents repression of the RUNX2 P1 promoter during myogenic differentiation of mesenchymal cells. The epigenetically forced expression of RUNX2/p57 and osteocalcin, a classical bone-related target gene, under myoblastic-differentiation is accompanied by enrichment of the H3K4me3 and H3K27ac marks at the RUNX2 P1 promoter region. Our results identify JARID1B as a key component of a potent epigenetic switch that controls mesenchymal cell fate into myogenic and osteogenic lineages.
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Oncogenic cooperation between PI3K/Akt signaling and Transcription Factor RUNX2 promotes the invasive properties of metastatic breast cancer cells
Journal of Cellular Physiology, 2013Co-Authors: Sandhya Pande, Sayyed K Zaidi, Jane B Lian, Andre J Van Wijnen, Janet L. Stein, Gillian Browne, Srivatsan Padmanabhan, Gary S. SteinAbstract:The serine/threonine kinase Akt/PKB promotes cancer cell growth and invasion through several downstream targets. Identification of novel substrates may provide new avenues for therapeutic intervention. Our study shows that Akt phosphorylates the cancer-related Transcription Factor RUNX2 resulting in stimulated DNA binding of the purified recombinant protein in vitro. Pharmacological inhibition of the PI3K/Akt pathway in breast cancer cells reduces DNA-binding activity of RUNX2 with concomitant reduction in the expression of metastasis-related RUNX2 target genes. Akt phosphorylates RUNX2 at three critical residues within the runt DNA-binding domain to enhance its in vivo genomic interactions with a target gene promoter, MMP13. Mutation of these three phosphorylation sites reduces RUNX2 DNA-binding activity. However, Akt signaling does not appear to interefere with CBFβ-RUNX2 interactions. Consequently, expression of multiple metastasis-related genes is decreased and RUNX2-mediated cell invasion is supressed. Thus, our work identifies RUNX2 as a novel and important downstream mediator of the PI3K/Akt pathway that is linked to metastatic properties of breast cancer cells. J. Cell. Physiol. 228: 1784–1792, 2013. © 2013 Wiley Periodicals, Inc.
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genomic promoter occupancy of runt related Transcription Factor RUNX2 in osteosarcoma cells identifies genes involved in cell adhesion and motility
Journal of Biological Chemistry, 2012Co-Authors: Margaretha Van Der Deen, Jacqueline Akech, Sneha Gupta, Martin Montecino, Jane B Lian, Mario Galindo, Daniel W Young, David S Lapointe, Janet L. Stein, Gary S. SteinAbstract:Abstract Runt-related Transcription Factors (RUNX1, RUNX2, and RUNX3) are key lineage-specific regulators of progenitor cell growth and differentiation but also function pathologically as cancer genes that contribute to tumorigenesis. RUNX2 attenuates growth and stimulates maturation of osteoblasts during bone formation but is also robustly expressed in a subset of osteosarcomas, as well as in metastatic breast and prostate tumors. To assess the biological function of RUNX2 in osteosarcoma cells, we examined human genomic promoter interactions for RUNX2 using chromatin immunoprecipitation (ChIP)-microarray analysis in SAOS-2 cells. Promoter binding of both RUNX2 and RNA polymerase II was compared with gene expression profiles of cells in which RUNX2 was depleted by RNA interference. Many RUNX2-bound loci (1550 of 2339 total) exhibit promoter occupancy by RNA polymerase II and contain the RUNX consensus motif 5′-((T/A/C)G(T/A/C)GG(T/G). Gene ontology analysis indicates that RUNX2 controls components of multiple signaling pathways (e.g. WNT, TGFβ, TNFα, and interleukins), as well as genes linked to cell motility and adhesion (e.g. the focal adhesion-related genes FAK/PTK2 and TLN1). Our results reveal that siRNA depletion of RUNX2, PTK2, or TLN1 diminishes motility of U2OS osteosarcoma cells. Thus, RUNX2 binding to diverse gene loci may support the biological properties of osteosarcoma cells.
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a program of micrornas controls osteogenic lineage progression by targeting Transcription Factor RUNX2
Proceedings of the National Academy of Sciences of the United States of America, 2011Co-Authors: Ying Zhang, Jane B Lian, Andre J Van Wijnen, Janet L. Stein, Carlo M. Croce, Gary S. SteinAbstract:Lineage progression in osteoblasts and chondrocytes is stringently controlled by the cell-fate–determining Transcription Factor RUNX2. In this study, we directly addressed whether microRNAs (miRNAs) can control the osteogenic activity of RUNX2 and affect osteoblast maturation. A panel of 11 RUNX2-targeting miRNAs (miR-23a, miR-30c, miR-34c, miR-133a, miR-135a, miR-137, miR-204, miR-205, miR-217, miR-218, and miR-338) is expressed in a lineage-related pattern in mesenchymal cell types. During both osteogenic and chondrogenic differentiation, these miRNAs, in general, are inversely expressed relative to RUNX2. Based on 3′UTR luciferase reporter, immunoblot, and mRNA stability assays, each miRNA directly attenuates RUNX2 protein accumulation. RUNX2-targeting miRNAs differentially inhibit RUNX2 protein expression in osteoblasts and chondrocytes and display different efficacies. Thus, cellular context contributes to miRNA-mediated regulation of RUNX2. All RUNX2-targeting miRNAs (except miR-218) significantly impede osteoblast differentiation, and their effects can be reversed by the corresponding anti-miRNAs. These findings demonstrate that osteoblastogenesis is limited by an elaborate network of functionally tested miRNAs that directly target the osteogenic master regulator RUNX2.
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live cell imaging of the cancer related Transcription Factor RUNX2 during mitotic progression
Journal of Cellular Physiology, 2011Co-Authors: Shirwin M Pockwinse, Jane B Lian, Andre J Van Wijnen, Gary S. Stein, Janet L. Stein, Krishna P Kota, Alexandre J C Quaresma, Anthony N Imbalzano, Jeffrey A NickersonAbstract:The nuclear matrix bound Transcription Factor RUNX2 is a lineage-specific developmental regulator that is linked to cancer. We have previously shown that RUNX2 controls Transcription of both RNA polymerase II genes and RNA polymerase I dependent ribosomal RNA genes. RUNX2 is epigenetically retained through mitosis on both classes of target genes in condensed chromosomes. We have used fluorescence recovery after photobleaching (FRAP) to measure the relative binding kinetics of EGFP-RUNX2 at Transcription sites in the nucleus and nucleoli during interphase, as well as on mitotic chromosomes. RUNX2 becomes more strongly bound as cells go from interphase through prophase, with a doubling of the most tightly bound “immobile fraction”. RUNX2 exchange then becomes much more facile during metaphase to telophase. During interphase the less tightly bound pool of RUNX2 exchanges more slowly at nucleoli than at subnuclear foci, and the non-exchanging immobile fraction is greater in nucleoli. These results are consistent with a model in which the molecular mechanism of RUNX2 binding is different at protein-coding and ribosomal RNA genes. The binding interactions of RUNX2 change as cells go through mitosis, with binding affinity increasing as chromosomes condense and then decreasing through subsequent mitotic phases. The increased residence of RUNX2 at mitotic chromosomes may reflect its epigenetic function in “bookmarking” of target genes in cancer cells.