DOCSLIB.ORG

Effect of Akt Inhibition on Scatter Factor-Regulated Gene Expression in DU-145 Human Prostate Cancer Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Effect of Akt Inhibition on Scatter Factor-Regulated Gene Expression in DU-145 Human Prostate Cancer Cells Oncogene (2007) 26, 2925–2938 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc ORIGINAL ARTICLE Effect of Akt inhibition on scatter factor-regulated gene expression in DU-145 human prostate cancer cells JXu1, M Gao2, S Fan1, Q Meng1, ID Goldberg2, R Abounader3, H Ressom1, JJ Laterra3 and EM Rosen1 1Department of Oncology, Lombardi Comprehensive Cancer Center/Georgetown University, Washington, DC, USA; 2Department of Radiation Oncology, Long Island Jewish Medical Center, The Long Island Campus for the Albert Einstein College of Medicine, New York, NY, USA and 3Department of Neurooncology, The Kennedy Krieger Institute/Johns Hopkins University School of Medicine, Baltimore, MD, USA The cytokine scatter factor (SF) (hepatocyte growth Introduction factor) transduces various biologic actions, including cell motility, invasion, angiogenesis and apoptosis inhibition. Scatter factor (SF) (hepatocyte growth factor) is a The latter is relevant to understanding the role of SF in pleiotrophic cytokine that regulates cell motility, inva- promoting tumor cell survival in different contexts, for sion, proliferation, morphogenesis, angiogenesis, tumori- example, detachment from basement membrane, growth genesis and transformation in different cell types and in metastatic sites and responses to chemo- and radio- contexts. These actions are transduced through c-Met, a therapy. Previously, we showed that SF protects cells receptor tyrosine kinase (Bottaro et al., 1991). SF can against apoptosis owing to DNA damage, by a mechanism protect various cell types against apoptosis owing to involving phosphoinositol-3-kinase/c-Akt signaling. Here, detachment from the substratum (anoikis) (Frisch and we used DNA microarray assays to identify c-Akt- Francis, 1994), autoimmune responses (Futamatsu regulated genes that might contribute to cell protection. et al., 2005), intracellular infection (Leiriao et al., DU-145 human prostate cancer cells were transfected7a 2005), staurosporine (Bardelli et al., 1996), the sphingo- dominant-negative mutant Akt, treated7SF and analysed lipid ceramide (Kannan et al., 2004), hypoxia/reperfu- for gene expression using Affymetrix arrays. These sion injury (Wang et al., 2004) and various other agents studies identified SF-regulated genes for which induction (Dworkin et al., 2004; Huh et al., 2004). Although SF is was c-Akt-dependent vs -independent. Selected micro- usually cytoprotective, it can also mediate proapoptotic array findings were confirmed by semiquantitative and effects through several mechanisms (Matteucci et al., quantitative reverse transcription–polymerase chain reac- 2003). tion. We tested the contribution of four SF-inducible/ We reported that SF protects epithelial, carcinoma c-Akt-dependent genes (AMPD3, EPHB2, MX1 and and glioma cells against apoptosis owing to DNA- WNT4) to protection against adriamycin (a DNA topo- damaging agents, including ionizing radiation and isomerase IIa inhibitor) using RNA interference. Knock- chemotherapy drugs, for example, adriamycin (ADR) down of each gene except EPHB2 caused a small but (a DNA topoisomerase IIa inhibitor) and cis-platinum significant reduction in the SF cell protection. The lack of (a crosslinking agent). This protection is due to effect of EPHB2 knockdown may be due to the fact that antiapoptotic signaling, inhibition of the mitochondrial DU-145 cells contain a single-mutant EPHB2 allele. A apoptosis pathway and stimulation of DNA strand combination of three small interfering RNAs blocked break repair (Fan et al., 1998, 2000, 2001; Bowers et al., most of the protection by SF in both DU-145 and T47D 2000; Gao et al., 2001). The protection of cancer cells cells. These findings identify novel c-Akt-regulated genes, against DNA damage-induced apoptosis is a clinically some of which contribute to SF-mediated cytoprotection. relevant problem because SF accumulates and c-Met is Oncogene (2007) 26, 2925–2938. doi:10.1038/sj.onc.1210088; often upregulated in various tumor types, including published online 13 November 2006 cancers of the breast, bladder, prostate and brain (gliomas) (Jin et al., 1997; Rosen et al., 1997; Beppu Keywords: scatter factor (SF); hepatocyte growth factor et al., 2000; Knudsen and Edlund, 2004). Thus, SF may (HGF); c-Akt; dominant-negative; gene expression; increase tumor cell resistance to radiation and chemo- DU-145 therapy by blocking DNA damage-induced apoptosis and stimulating DNA repair. We have studied the mechanisms of SF-mediated cell Correspondence: Dr EM Rosen, Department of Oncology, Lombardi protection in several cell types, including MDCK canine Comprehensive Cancer Center/Georgetown University, 3970 Reser- epithelial cells, human breast and prostate cancer cells, voir Road, NW, Box 571469, Washington, DC 20057-1469, USA. E-mail: [email protected] and human and rat glioma cells. We found that SF Received 15 August 2005; revised 1 August 2006; accepted 1 August stimulates antiapoptotic signaling from the c-Met 2006; published online 13 November 2006 receptor to phosphatidylinositol-30-kinase (PI3K) and Effect of Akt on scatter factor-regulated gene expression JXuet al 2926 the cell survival-promoting serine/threonine kinase Condition 4 vs 3 (4/3): (SF þ DN-Akt)/DN-Akt c-Akt (Bowers et al., 2000; Fan et al., 2000, 2001). The SF-inducible genes with Akt inhibited multisubstrate adapter Gab1 (Grb2-associated binder-1) and the tumor suppressor PTEN (phosphatase and Genes on both lists are SF-inducible genes whose tensin homolog) act as upstream inhibitors of c-Akt to induction does not require c-Akt, whereas those on the block SF-mediated cell protection, whereas Pak1 (p21- first list but not the second are candidate genes whose associated kinase-1) acts downstream of c-Akt to induction by SF requires c-Akt. mediate cell protection (Fan et al., 2001, 2005). SF- Gene expression alterations were considered to be mediated activation of nuclear factor (NF-kB) signaling significant if: (1) the log (base 2) signal ratios were requires c-Src, c-Akt and Pak1, and NF-kB signaling is X þ 0.8 (ratio ¼ 1.74) or pÀ0.8 (ratio ¼ 0.57) in at least required for cell protection (Fan et al., 2005). two out of three independent experiments and (2) the The purpose of this study was to identify downstream corresponding P-values were significant, using the transcriptional targets of SF that are regulated through Affymetrix algorithm. The results for the 6 and 48 h c-Akt and contribute to cell protection. These studies time points are summarized in Table 1. At 6 h, 49 genes were carried out using DU-145 prostate cancer cells, a exhibited SF-induced alterations in expression based on cell type that we utilized in previous studies of c-Met the above criteria. Among these, 19 genes (40%) were no antiapoptosis signaling. longer altered in expression when DN-Akt was used to block c-Akt signaling. At 48 h, 104 genes showed SF-induced changes in expression and 44 of these Results genes (42%) were no longer changed in the presence of DN-Akt. At 48 h, we also observed a group of genes Identification of SF- and c-Akt-regulated genes in (N ¼ 114) whose expression was altered by SF only in DU-145 cells the presence of DN-Akt. Most of these genes (90%) The goal of this study was to identify SF-inducible genes were decreased in expression. This group may represent whose expression is dependent upon c-Akt signaling. genes for which c-Akt normally blocks the ability of SF The approach was to compare the gene expression to inhibit expression. profiles of cells treated with SF in the absence or The complete lists of genes altered by SF and/or presence of a dominant-negative (DN)-Akt expression DN-Akt and the magnitudes of the alterations are provi- vector. Briefly, subconfluent proliferating cells were ded in the Supplementary data (Supplementary Tables transfected with an empty vector (pcDNA3) or DN- S1–S9). Genes similarly altered by SF at T ¼ 6h in Akt and incubated7SF (100 ng/ml) for T ¼ 6or48h. the absence vs presence of DN-Akt (SF-regulated/ Four treatment conditions were tested: (1) pcDNA3 c-Akt-independent genes) are listed in Supplementary alone (control); (2) (SF þ pcDNA3); (3) DN-Akt alone Table S1, whereas the SF-regulated/c-Akt-dependent and (4) (DN-Akt þ SF). DNA microarray analyses were genes are listed in Supplementary Table S2. The performed using the Affymetrix HG-U133A gene chips. corresponding gene lists for T ¼ 48 h are shown in To identify SF-regulated genes that are dependent upon Supplementary Tables S3 and S4. Supplementary Table c-Akt, we compared the following: S2 includes some genes that were induced by SF in the absence of DN-Akt, but whose expression was Condition 2 vs 1 (2/1): (SF þ pcDNA3)/pcDNA3 decreased by SF in DN-Akt-transfected cells in one SF-inducible genes or more experiments. At 48 h, 60 genes showed similar Table 1 Summary of results from DNA microarray experimentsa Table number Comparison Gene subset Number Number of genes of genes increased decreased S1 [pcDNA3+SF]/pcDNA3, Similarly regulated in the presence and absence of DN-Akt 29 1 [DN-Akt+SF]/DN-Akt (T ¼ 6h) S2 [pcDNA3+SF]/pcDNA3, Altered in the absence but not presence of DN-Akt 18 1 [DN-Akt+SF]/DN-Akt (T ¼ 6h) S3 [pcDNA3+SF]/pcDNA3, Similarly regulated in the presence and absence of DN-Akt 40 20 [DN-Akt+SF]/DN-Akt (T ¼ 48 h) S4 [pcDNA3+SF]/pcDNA3, Altered in the absence but not presence of DN-Akt 13 31 [DN-Akt+SF]/DN-Akt (T ¼ 48 h) S5 [pcDNA3+SF]/pcDNA3, Altered in the presence but not absence of DN-Akt 11 103 [DN-Akt+SF]/DN-Akt (T ¼ 48 h) S6 DN-Akt vs pcDNA3 (T ¼ 6 h) All 27 — S7 DN-Akt vs pcDNA3 (T ¼ 48 h) All 9 — S8 [DN-Akt+SF] vs [pcDNA3+SF] (T ¼ 6 h) All 8 — S9 [DN-Akt+SF] vs [pcDNA3+SF] (T ¼ 48 h) All 12 5 Abbreviations: DN-Akt, dominant-negative mutant Akt; SF, scatter factor.
Load more

Recommended publications
  • The Role and Mechanisms of Action of Micrornas in Cancer Drug Resistance Wengong Si1,2,3, Jiaying Shen4, Huilin Zheng1,5 and Weimin Fan1,6*
    Si et al. Clinical Epigenetics (2019) 11:25 https://doi.org/10.1186/s13148-018-0587-8 REVIEW Open Access The role and mechanisms of action of microRNAs in cancer drug resistance Wengong Si1,2,3, Jiaying Shen4, Huilin Zheng1,5 and Weimin Fan1,6* Abstract MicroRNAs (miRNAs) are small non-coding RNAs with a length of about 19–25 nt, which can regulate various target genes and are thus involved in the regulation of a variety of biological and pathological processes, including the formation and development of cancer. Drug resistance in cancer chemotherapy is one of the main obstacles to curing this malignant disease. Statistical data indicate that over 90% of the mortality of patients with cancer is related to drug resistance. Drug resistance of cancer chemotherapy can be caused by many mechanisms, such as decreased antitumor drug uptake, modified drug targets, altered cell cycle checkpoints, or increased DNA damage repair, among others. In recent years, many studies have shown that miRNAs are involved in the drug resistance of tumor cells by targeting drug-resistance-related genes or influencing genes related to cell proliferation, cell cycle, and apoptosis. A single miRNA often targets a number of genes, and its regulatory effect is tissue-specific. In this review, we emphasize the miRNAs that are involved in the regulation of drug resistance among different cancers and probe the mechanisms of the deregulated expression of miRNAs. The molecular targets of miRNAs and their underlying signaling pathways are also explored comprehensively. A holistic understanding of the functions of miRNAs in drug resistance will help us develop better strategies to regulate them efficiently and will finally pave the way toward better translation of miRNAs into clinics, developing them into a promising approach in cancer therapy.
    [Show full text]
  • Principles and Applications of Rabbit Models for Atherosclerosis Research
    J Atheroscler Thromb, 2018; 25: 213-220. http://doi.org/10.5551/jat.RV17018 Review Principles and Applications of Rabbit Models for Atherosclerosis Research Jianglin Fan1, Yajie Chen1, Haizhao Yan1, Manabu Niimi1, Yanli Wang2 and Jingyan Liang3 1Department of Molecular Pathology, Faculty of Medicine, Graduate School of Medical Sciences, University of Yamanashi, Yamanashi, Japan 2Department of Pathology, Xi’an Medical University, Xi’an, China 3Research Center for Vascular Biology, Yangzhou University School of Medicine, Yangzhou, China Rabbits are one of the most used experimental animals for biomedical research, particularly as a bio- reactor for the production of antibodies. However, many unique features of the rabbit have also made it as an excellent species for examining a number of aspects of human diseases such as atherosclerosis. Rabbits are phylogenetically closer to humans than rodents, in addition to their relatively proper size, tame disposition, and ease of use and maintenance in the laboratory facility. Due to their short life spans, short gestation periods, high numbers of progeny, low cost (compared with other large ani- mals) and availability of genomics and proteomics, rabbits usually serve to bridge the gap between smaller rodents (mice and rats) and larger animals, such as dogs, pigs and monkeys, and play an important role in many translational research activities such as pre-clinical testing of drugs and diag- nostic methods for patients. The principle of using rabbits rather than other animals as an experi- mental model is very simple: rabbits should be used for research, such as translational research, that is difficult to accomplish with other species. Recently, rabbit genome sequencing and transcriptomic profiling of atherosclerosis have been successfully completed, which has paved a new way for researchers to use this model in the future.
    [Show full text]
  • WO 2017/147196 Al 31 August 2017 (31.08.2017) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/147196 Al 31 August 2017 (31.08.2017) P O P C T (51) International Patent Classification: Kellie, E. [US/US]; 70 Lanark Road, Maiden, MA 02148 C12Q 1/68 (2006.01) (US). COLE, Michael, B. [US/US]; 233 1 Eunice Street, Berkeley, CA 94708 (US). YOSEF, Nir [IL/US]; 1520 (21) International Application Number: Laurel Ave., Richmond, CA 94805 (US). GAYO, En¬ PCT/US20 17/0 18963 rique, Martin [ES/US]; 115 Peterborough Street, Boston, (22) International Filing Date: MA 022 15 (US). OUYANG, Zhengyu [CN/US]; 15 Vas- 22 February 2017 (22.02.2017) sar Street, Medford, MA 02155 (US). YU, Xu [CN/US]; 6 Whittier Place, Apt. 16j, Boston, MA 02 114 (US). (25) Filing Language: English (74) Agents: KOWALSKI, Thomas, J. et al; Vedder Price English (26) Publication Language: P.C., 1633 Broadway, New York, NY 1001 9 (US). (30) Priority Data: (81) Designated States (unless otherwise indicated, for every 62/298,349 22 February 2016 (22.02.2016) US kind of national protection available): AE, AG, AL, AM, (71) Applicants: MASSACHUSETTS INSTITUTE OF AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, TECHNOLOGY [US/US]; 77 Massachusetts Ave., Cam BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, bridge, MA 02139 (US). THE REGENTS OF THE UNI¬ DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, VERSITY OF CALIFORNIA [US/US]; 1111 Franklin HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, Street, 12th Floor, Oakland, CA 94607 (US).
    [Show full text]
  • Expression Profiling of KLF4
    Expression Profiling of KLF4 AJCR0000006 Supplemental Data Figure S1. Snapshot of enriched gene sets identified by GSEA in Klf4-null MEFs. Figure S2. Snapshot of enriched gene sets identified by GSEA in wild type MEFs. 98 Am J Cancer Res 2011;1(1):85-97 Table S1: Functional Annotation Clustering of Genes Up-Regulated in Klf4 -Null MEFs ILLUMINA_ID Gene Symbol Gene Name (Description) P -value Fold-Change Cell Cycle 8.00E-03 ILMN_1217331 Mcm6 MINICHROMOSOME MAINTENANCE DEFICIENT 6 40.36 ILMN_2723931 E2f6 E2F TRANSCRIPTION FACTOR 6 26.8 ILMN_2724570 Mapk12 MITOGEN-ACTIVATED PROTEIN KINASE 12 22.19 ILMN_1218470 Cdk2 CYCLIN-DEPENDENT KINASE 2 9.32 ILMN_1234909 Tipin TIMELESS INTERACTING PROTEIN 5.3 ILMN_1212692 Mapk13 SAPK/ERK/KINASE 4 4.96 ILMN_2666690 Cul7 CULLIN 7 2.23 ILMN_2681776 Mapk6 MITOGEN ACTIVATED PROTEIN KINASE 4 2.11 ILMN_2652909 Ddit3 DNA-DAMAGE INDUCIBLE TRANSCRIPT 3 2.07 ILMN_2742152 Gadd45a GROWTH ARREST AND DNA-DAMAGE-INDUCIBLE 45 ALPHA 1.92 ILMN_1212787 Pttg1 PITUITARY TUMOR-TRANSFORMING 1 1.8 ILMN_1216721 Cdk5 CYCLIN-DEPENDENT KINASE 5 1.78 ILMN_1227009 Gas2l1 GROWTH ARREST-SPECIFIC 2 LIKE 1 1.74 ILMN_2663009 Rassf5 RAS ASSOCIATION (RALGDS/AF-6) DOMAIN FAMILY 5 1.64 ILMN_1220454 Anapc13 ANAPHASE PROMOTING COMPLEX SUBUNIT 13 1.61 ILMN_1216213 Incenp INNER CENTROMERE PROTEIN 1.56 ILMN_1256301 Rcc2 REGULATOR OF CHROMOSOME CONDENSATION 2 1.53 Extracellular Matrix 5.80E-06 ILMN_2735184 Col18a1 PROCOLLAGEN, TYPE XVIII, ALPHA 1 51.5 ILMN_1223997 Crtap CARTILAGE ASSOCIATED PROTEIN 32.74 ILMN_2753809 Mmp3 MATRIX METALLOPEPTIDASE
    [Show full text]
  • Annals of Medical and Clinical Oncology Chen C, Et Al
    Annals of Medical and Clinical Oncology Chen C, et al. Ann med clin Oncol 3: 125. Short Commentary DOI: 10.29011/AMCO-125.000125 Commentary Referring to Pericyte FAK Negatively Regulates Gas6/ Axl Signalling To Suppress Tumour Angiogenesis and Tumour Growth Chen Chen, Hongyan Wang, Binfeng Wu and Jinghua Chen* Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, PR China *Corresponding author: Jinghua Chen, Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, PR China Citation: Chen C, Wang H, Wu B, Chen J (2020) Commentary Referring to Pericyte FAK Negatively Regulates Gas6/Axl Signalling To Suppress Tumour Angiogenesis and Tumour Growth. Ann med clin Oncol 3: 125. DOI: 10.29011/AMCO-125.000125 Received Date: 07 December, 2020; Accepted Date: 20 December, 2020; Published Date: 28 December, 2020 The published research article utilized multiple mouse Interestingly, knockout of FAK was specific to pericytes other models including melanoma, lung carcinoma and pancreatic B-cell than endothelial cells, mice models demonstrated that loss of insulinoma. Two hallmarks of cancer [1] such as angiogenesis and FAK from pericytes significantly promoted ɑ-SMA expression tumour growth had been evaluated. Two major molecules FAK (common metastatic biomarker) and NG-2 (typical angiogenesis (focal adhesion kinase 1) and Axl undertook the innovative roles related biomarker) in three cancer cells model. It suggested that of this elegant paper. They both belong to protein Tyrosine Kinase FAK may function as the tumour suppressive gene. However, (TK) family [2].
    [Show full text]
  • Cysteine-Rich 61 (Cyr61): a Biomarker Reflecting Disease Activity In
    Fan et al. Arthritis Research & Therapy (2019) 21:123 https://doi.org/10.1186/s13075-019-1906-y RESEARCHARTICLE Open Access Cysteine-rich 61 (Cyr61): a biomarker reflecting disease activity in rheumatoid arthritis Yong Fan†, Xinlei Yang†, Juan Zhao, Xiaoying Sun, Wenhui Xie, Yanrong Huang, Guangtao Li, Yanjie Hao and Zhuoli Zhang* Abstract Background: Numerous preclinical studies have revealed a critical role of cysteine-rich 61 (Cyr61) in the pathogenesis of rheumatoid arthritis (RA). But there is little literature discussing the clinical value of circulation Cyr61 in RA patients. The aim of our study is to investigate the serum Cyr61 level and its association with disease activity in RA patients. Methods: A training cohort was derived from consecutive RA patients who visited our clinic from Jun 2014 to Nov 2018. Serum samples were obtained at the enrollment time. To further confirm discovery, an independent validation cohort was set up based on a registered clinical trial. Paired serum samples of active RA patients were respectively collected at baseline and 12 weeks after uniformed treatment. Serum Cyr61 concentration was detected by enzyme-linked immunosorbent assay. The comparison of Cyr61 between RA patients and controls, the correlation between Cyr61 levels with disease activity, and the change of Cyr61 after treatment were analyzed by appropriate statistical analyses. Results: A total of 177 definite RA patients and 50 age- and gender-matched healthy controls were enrolled in the training cohort. Significantly elevated serum Cyr61 concentration was found in RA patients, demonstrating excellent diagnostic ability to discriminate RA from healthy controls (area under the curve (AUC) = 0.98, P < 0.001).
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • Salt-Inducible Kinases Dictate Parathyroid Hormone Receptor Action in Bone Development and Remodeling
    Salt-inducible kinases dictate parathyroid hormone receptor action in bone development and remodeling Shigeki Nishimori, … , Henry M. Kronenberg, Marc N. Wein J Clin Invest. 2019. https://doi.org/10.1172/JCI130126. Research In-Press Preview Bone Biology Endocrinology The parathyroid hormone receptor (PTH1R) mediates the biologic actions of parathyroid hormone (PTH) and parathyroid hormone related protein (PTHrP). Here, we showed that salt inducible kinases (SIKs) are key kinases that control the skeletal actions downstream of PTH1R and that this GPCR, when activated, inhibited cellular SIK activity. Sik gene deletion led to phenotypic changes that were remarkably similar to models of increased PTH1R signaling. In growth plate chondrocytes, PTHrP inhibited SIK3 and ablation of this kinase in proliferating chondrocytes rescued perinatal lethality of PTHrP-null mice. Combined deletion of Sik2/Sik3 in osteoblasts and osteocytes led to a dramatic increase in bone mass that closely resembled the skeletal and molecular phenotypes observed when these bone cells express a constitutively active PTH1R that causes Jansen’s metaphyseal chondrodysplasia. Finally, genetic evidence demonstrated that class IIa HDACs were key PTH1R-regulated SIK substrates in both chondrocytes and osteocytes. Taken together, our findings established that SIK inhibition is central to PTH1R action in bone development and remodeling. Furthermore, this work highlighted the key role of cAMP-regulated salt inducible kinases downstream of GPCR action. Find the latest version: https://jci.me/130126/pdf 1 Salt-inducible kinases dictate parathyroid hormone receptor action in bone development 2 and remodeling 3 4 Shigeki Nishimori1,2, Maureen J. O’Meara1, Christian Castro1, Hiroshi Noda1,3, Murat Cetinbas4, 5 Janaina da Silva Martins1, Ugur Ayturk5, Daniel J.
    [Show full text]
  • Test Catalogue August 2019
    Test Catalogue August 2019 www.centogene.com/catalogue Table of Contents CENTOGENE CLINICAL DIAGNOSTIC PRODUCTS AND SERVICES › Whole Exome Testing 4 › Whole Genome Testing 5 › Genome wide CNV Analysis 5 › Somatic Mutation Analyses 5 › Biomarker Testing, Biochemical Testing 6 › Prenatal Testing 7 › Additional Services 7 › Metabolic Diseases 9 - 21 › Neurological Diseases 23 - 47 › Ophthalmological Diseases 49 - 55 › Ear, Nose and Throat Diseases 57 - 61 › Bone, Skin and Immune Diseases 63 - 73 › Cardiological Diseases 75 - 79 › Vascular Diseases 81 - 82 › Liver, Kidney and Endocrinological Diseases 83 - 89 › Reproductive Genetics 91 › Haematological Diseases 93 - 96 › Malformation and/or Retardation Syndromes 97 - 107 › Oncogenetics 109 - 113 ® › CentoXome - Sequencing targeting exonic regions of ~20.000 genes Test Test name Description code CentoXome® Solo Medical interpretation/report of WES findings for index 50029 CentoXome® Solo - Variants Raw data; fastQ, BAM, Vcf files along with variant annotated file in xls format for index 50028 CentoXome® Solo - with CNV Medical interpretation/report of WES including CNV findings for index 50103 Medical interpretation/report of WES in index, package including genome wide analyses of structural/ CentoXome® Solo - with sWGS 50104 large CNVs through sWGS Medical interpretation/report of WES in index, package including genome wide analyses of structural/ CentoXome® Solo - with aCGH 750k 50122 large CNVs through 750k microarray Medical interpretation/report of WES in index, package including genome
    [Show full text]
  • ETV5 Links the FGFR3 and Hippo Signalling Pathways in Bladder Cancer Received: 2 December 2016 Erica Di Martino, Olivia Alder , Carolyn D
    www.nature.com/scientificreports OPEN ETV5 links the FGFR3 and Hippo signalling pathways in bladder cancer Received: 2 December 2016 Erica di Martino, Olivia Alder , Carolyn D. Hurst & Margaret A. Knowles Accepted: 14 November 2018 Activating mutations of fbroblast growth factor receptor 3 (FGFR3) are common in urothelial Published: xx xx xxxx carcinoma of the bladder (UC). Silencing or inhibition of mutant FGFR3 in bladder cancer cell lines is associated with decreased malignant potential, confrming its important driver role in UC. However, understanding of how FGFR3 activation drives urothelial malignant transformation remains limited. We have previously shown that mutant FGFR3 alters the cell-cell and cell-matrix adhesion properties of urothelial cells, resulting in loss of contact-inhibition of proliferation. In this study, we investigate a transcription factor of the ETS-family, ETV5, as a putative efector of FGFR3 signalling in bladder cancer. We show that FGFR3 signalling induces a MAPK/ERK-mediated increase in ETV5 levels, and that this results in increased level of TAZ, a co-transcriptional regulator downstream of the Hippo signalling pathway involved in cell-contact inhibition. We also demonstrate that ETV5 is a key downstream mediator of the oncogenic efects of mutant FGFR3, as its knockdown in FGFR3-mutant bladder cancer cell lines is associated with reduced proliferation and anchorage-independent growth. Overall this study advances our understanding of the molecular alterations occurring during urothelial malignant transformation and indicates TAZ as a possible therapeutic target in FGFR3-dependent bladder tumours. Fibroblast growth factors (FGF) and their four tyrosine kinase receptors (FGFR1-4) activate multiple downstream cellular signalling pathways, such as MAPK/ERK, PLCγ1, PI3K and STATs, and regulate a variety of physiolog- ical processes, encompassing embryogenesis, angiogenesis, metabolism, and wound healing1–3.
    [Show full text]
  • Eyeing the Cyr61/CTGF/NOV (CCN) Group of Genes in Development And
    Krupska et al. Human Genomics (2015) 9:24 DOI 10.1186/s40246-015-0046-y REVIEW Open Access Eyeing the Cyr61/CTGF/NOV (CCN) group of genes in development and diseases: highlights of their structural likenesses and functional dissimilarities Izabela Krupska1,3, Elspeth A. Bruford2 and Brahim Chaqour1,3,4* Abstract “CCN” is an acronym referring to the first letter of each of the first three members of this original group of mammalian functionally and phylogenetically distinct extracellular matrix (ECM) proteins [i.e., cysteine-rich 61 (CYR61), connective tissue growth factor (CTGF), and nephroblastoma-overexpressed (NOV)]. Although “CCN” genes are unlikely to have arisen from a common ancestral gene, their encoded proteins share multimodular structures in which most cysteine residues are strictly conserved in their positions within several structural motifs. The CCN genes can be subdivided into members developmentally indispensable for embryonic viability (e.g., CCN1, 2 and 5), each assuming unique tissue-specific functions, and members not essential for embryonic development (e.g., CCN3, 4 and 6), probably due to a balance of functional redundancy and specialization during evolution. The temporo-spatial regulation of the CCN genes and the structural information contained within the sequences of their encoded proteins reflect diversity in their context and tissue-specific functions. Genetic association studies and experimental anomalies, replicated in various animal models, have shown that altered CCN gene structure or expression is associated with “injury” stimuli—whether mechanical (e.g., trauma, shear stress) or chemical (e.g., ischemia, hyperglycemia, hyperlipidemia, inflammation). Consequently, increased organ-specific susceptibility to structural damages ensues. These data underscore the critical functions of CCN proteins in the dynamics of tissue repair and regeneration and in the compensatory responses preceding organ failure.
    [Show full text]
  • Enhancing the Astrocytic Clearance of Extracellular Α-Synuclein
    Cellular and Molecular Neurobiology (2019) 39:1017–1028 https://doi.org/10.1007/s10571-019-00696-2 ORIGINAL RESEARCH Enhancing the Astrocytic Clearance of Extracellular α‑Synuclein Aggregates by Ginkgolides Attenuates Neural Cell Injury Jun Hua1,2 · Nuo Yin1 · Shi Xu1 · Qiang Chen1,3 · Tingting Tao1,3 · Ji Zhang4 · Jianhua Ding1 · Yi Fan1,3 · Gang Hu1,5 Received: 9 November 2018 / Accepted: 1 June 2019 / Published online: 5 June 2019 © Springer Science+Business Media, LLC, part of Springer Nature 2019 Abstract The accumulation of aggregated forms of the α-Synuclein (α-Syn) is associated with the pathogenesis of Parkinson’s dis- ease (PD), and the efcient clearance of aggregated α-Syn represents a potential approach in PD therapy. Astrocytes are the most numerous glia cells in the brain and play an essential role in supporting brain functions in PD state. In the present study, we demonstrated that cultured primary astrocytes engulfed and degraded extracellular aggregated recombinant human α-Syn. Meanwhile, we observed that the clearance of α-Syn by astrocytes was abolished by proteasome inhibitor MG132 and autophagy inhibitor 3-methyladenine (3MA). We further showed that intracellular α-Syn was reduced after ginkgolide B (GB) and bilobalide (BB) treatment, and the decrease was reversed by MG132 and 3MA. More importantly, GB and BB reduced indirect neurotoxicity to neurons induced by α-Syn-stimulated astrocytic conditioned medium. Together, we frstly fnd that astrocytes can engulf and degrade α-Syn aggregates via the proteasome and autophagy pathways, and further show that GB and BB enhance astrocytic clearance of α-Syn, which gives us an insight into the novel therapy for PD in future.
    [Show full text]