DOCSLIB.ORG

APC Mutations As a Potential Biomarker for Sensitivity To

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Published OnlineFirst February 8, 2017; DOI: 10.1158/1535-7163.MCT-16-0578 Companion Diagnostics and Cancer Biomarkers Molecular Cancer Therapeutics APC Mutations as a Potential Biomarker for Sensitivity to Tankyrase Inhibitors in Colorectal Cancer Noritaka Tanaka1,2, Tetsuo Mashima1, Anna Mizutani1, Ayana Sato1,3, Aki Aoyama3,4, Bo Gong3,4, Haruka Yoshida1, Yukiko Muramatsu1, Kento Nakata1,5, Masaaki Matsuura6, Ryohei Katayama4, Satoshi Nagayama7, Naoya Fujita3,4,5, Yoshikazu Sugimoto2, and Hiroyuki Seimiya1,3,5 Abstract In most colorectal cancers, Wnt/b-catenin signaling is acti- "short" truncated APCs lacking all seven b-catenin-binding vated by loss-of-function mutations in the adenomatous polyposis 20-amino acid repeats (20-AARs). In contrast, the drug-resistant coli (APC) gene and plays a critical role in tumorigenesis. cells possessed "long" APC retaining two or more 20-AARs. Knock- Tankyrases poly(ADP-ribosyl)ate and destabilize Axins, a neg- down of the long APCs with two 20-AARs increased b-catenin, ative regulator of b-catenin, and upregulate b-catenin signaling. Tcf/LEF transcriptional activity and its target gene AXIN2 expres- Tankyrase inhibitors downregulate b-catenin and are expected sion. Under these conditions, tankyrase inhibitors were able to to be promising therapeutics for colorectal cancer. However, downregulate b-catenin in the resistant cells. These results indicate colorectal cancer cells are not always sensitive to tankyrase that the long APCs are hypomorphic mutants, whereas they exert inhibitors, and predictive biomarkers for the drug sensitivity a dominant-negative effect on Axin-dependent b-catenin degra- remain elusive. Here we demonstrate that the short-form APC dation caused by tankyrase inhibitors. Finally, we established mutations predict the sensitivity of colorectal cancer cells to 16 patient-derived colorectal cancer cells and confirmed that tankyrase inhibitors. By using well-established colorectal cancer the tankyrase inhibitor–responsive cells harbor the short-form cell lines, we found that tankyrase inhibitors downregulated APC mutations. These observations exemplify the predictive b-catenin in the drug-sensitive, but not resistant, colorectal importance of APC mutations, the most common genetic alter- cancer cells. The drug-sensitive cells showed higher Tcf/LEF ation in colorectal cancers, for molecular targeted therapeutics. transcriptional activity than the resistant cells and possessed Mol Cancer Ther; 16(4); 752–62. Ó2017 AACR. Introduction those in KRAS, SMAD4, and TP53 (3, 4). APC functions as a functional scaffold for the b-catenin destruction complex, which Loss-of-function mutations in adenomatous polyposis coli (APC) is composed of b-catenin, Axin, casein kinase 1a and glycogen gene, a negative regulator of Wnt/b-catenin signaling, frequently synthase kinase 3b (GSK3b), and suppresses Wnt/b-catenin occur in colorectal cancer (1, 2). These APC mutations promote signaling (5–10). In the destruction complex, APC and Axin tumorigenesis together with several common mutations, such as promote the ubiquitin-dependent proteasomal degradation of b-catenin via casein kinase 1a- and GSK3b-mediated phosphory- 1Division of Molecular Biotherapy, Cancer Chemotherapy Center, Japanese lation of b-catenin (8, 10). In genetically engineered mouse Foundation for Cancer Research, Tokyo, Japan. 2Division of Chemotherapy, models, APC mutation–derived activation of Wnt/b-catenin sig- 3 Faculty of Pharmacy, Keio University, Tokyo, Japan. Laboratory of Molecular naling was shown to be required not only for promotion of Target Therapy of Cancer, Department of Computational Biology and Medical colorectal cancer but also for tumor maintenance (11, 12). These Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, b Japan. 4Division of Experimental Chemotherapy, Cancer Chemotherapy Center, observations indicate that the Wnt/ -catenin signaling pathway Japanese Foundation for Cancer Research, Tokyo, Japan. 5Graduate School of is a rational therapeutic target for colorectal cancer. So far, how- Pharmaceutical Sciences, Meiji Pharmaceutical University, Tokyo, Japan. 6Grad- ever, this pathway lacks druggable molecular targets, which has uate School of Public Health, Teikyo University, Tokyo, Japan. 7Department of hampered the development of therapeutic drugs targeting Gastroenterological and Surgery, Cancer Institute Hospital, Japanese Founda- Wnt/b-catenin signaling. tion for Cancer Research, Tokyo, Japan. Tankyrases (tankyrase-1/PARP-5a and tankyrase-2/PARP-5b) Note: Supplementary data for this article are available at Molecular Cancer are members of the PARP family (13–15). This unique class of Therapeutics Online (http://mct.aacrjournals.org/). PARPs recognizes its binding partners via the multiple ankyrin Corresponding Author: Hiroyuki Seimiya, Division of Molecular Biotherapy, repeat cluster domain and poly(ADP-ribosyl)ates (PARylates) Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, them via the carboxyl terminal catalytic domain (13, 16, 17). 3-8-31 Ariake, Koto-ku, Tokyo 135-8550, Japan. Phone: 81-3-3570-0466; Fax: Tankyrases enhance Wnt/b-catenin signaling by PARylation and 81-3-3570-0484; E-mail: [email protected] subsequent destabilization of Axins (18, 19). Pharmacologic doi: 10.1158/1535-7163.MCT-16-0578 inhibition of tankyrases causes accumulation of Axins, which in Ó2017 American Association for Cancer Research. turn facilitates degradation of b-catenin and suppresses the 752 Mol Cancer Ther; 16(4) April 2017 Downloaded from mct.aacrjournals.org on September 24, 2021. © 2017 American Association for Cancer Research. Published OnlineFirst February 8, 2017; DOI: 10.1158/1535-7163.MCT-16-0578 APC as a Predictive Biomarker in Colorectal Cancer Therapy growth of several APC-mutated colorectal cancer cell lines antibodies were anti-tankyrase-1/2 (H350; Santa Cruz Biotech- (18, 20–23). These observations suggest that tankyrase inhibitors nology), anti-active b-catenin (8E7; Millipore), anti-phospho- are promising drugs for colorectal cancer treatment. However, b-catenin (Ser33/37/Thr41; Cell Signaling Technology), anti- colorectal cancer cells are not equally sensitive to tankyrase Axin1 (C76H11; Cell Signaling Technology), anti-Axin2 (76G6; inhibitors (22). Although appropriate stratification of patients Cell Signaling Technology), anti-GAPDH (6C5; Fitzgerald), anti- has profound implications for new drug development, predictive APC (ALi 12-28; Santa Cruz Biotechnology) for detecting the biomarkers for sensitivity to tankyrase inhibitors remain elusive. short APC (aa 1–811), and anti-APC (2504; Cell Signaling In this study, we demonstrate that a class of APC mutations Technology) for APC larger than 811 aa. predicts the sensitivity of colorectal cancer cells to tankyrase inhibitors. siRNA transfection ON-TARGETplus (human CTNNB1 and nontargeting control) APC Materials and Methods and Silencer Select [ (s1433: siAPC#1; s1434: siAPC#2; s1435: siAPC#3) and negative control no. 1] siRNAs were pur- Cell culture and proliferation assay chased from GE Healthcare Dharmacon and Thermo Fisher Human colorectal cancer cell lines COLO-320DM, SW403, Scientific, respectively. siRNAs were introduced into cells with SW480, and DLD-1 were maintained in RPMI1640 medium reverse transfection method using Lipofectamine RNAiMAX (Gibco, Life Technologies) supplemented with 10% FBS. RKO Transfection Reagent (Thermo Fisher Scientific). cells were maintained in DMEM low glucose (Nacalai Tesque) with 10% FBS. These cell lines were obtained from ATCC. HCT-15, qRT-PCR HCC2998, HT-29, KM12, and HCT-116 cells were maintained as Total RNAs were purified with the RNeasy Mini Kit (Qiagen) and described previously (24). All cell lines were authenticated by cDNA were synthesized with the SuperScript III First-Strand Syn- short tandem repeat profiling analysis (BEX) in 2016. Cell pro- thesis SuperMix for qRT-PCR (Thermo Fisher Scientific). The liferation was evaluated by the MTT assay (5-day drug treatment) expression levels of AXIN2 or Naked1 (NKD1)mRNAwerequan- as previously described (25) or by colony formation assay (10-day tified by real-time PCR analysis with the LightCycler 480 Real-Time drug treatment). Normalization was done to cells treated with PCR System with Universal ProbeLibrary Probe #36 (Roche). 0 DMSO as vehicle, which were defined as 100%. Final concentra- The primers were as follows: AXIN2,5-CACACCCTTCTCCAATC- 0 0 0 tion of DMSO was constant and the same for all the treatments. CAA-3 (forward) and 5 -TGCCAGTTTCTTTGGCTCTT-3 (reverse); and NKD1,50-TCTCGCCGGGATAGAAAAC-30 (forward) and 50- 0 Plasmid construction TCTCGCCGGGATAGAAAAC-3 (reverse). Total RNA was purified from COLO-320DM cells using the Immunofluorescence staining RNeasy Kit (Qiagen) and used as a template to amplify the Cells on glass coverslips were fixed with 2% formaldehyde/PBS truncated APC [amino acid (aa) 1–811] fragment by reverse and permeabilized with 0.5% Nonidet P-40/PBS. Immunofluo- transcription-PCR. The primer sets were: 50-ATCGGATCCGCT- rescence staining was performed as previously described (17) with GCCACCATGGAACAAAAGCTGATTTCTGAAGAAGATCTGGAA- antibody against nonphosphorylated (i.e., active) b-catenin TTCGCTGCAGCTTCATATGATCAGTTGTTA-30 (forward) and 50- 0 (D13A1; Cell Signaling Technology). The nuclei were stained ACACTCGAGATATGGTGAAAGGACAGTCATGTTGC-3 (reverse). 0 with VECTASHIELD Antifade Mounting Medium with 4 ,6-dia- The forward and reverse primers contain BamHI and XhoIsites, midino-2-phenylindole
Recommended publications
  • Rat Anti-TERF1 Monoclonal Antibody, Clone 683D (CABT-RM172) This Product Is for Research Use Only and Is Not Intended for Diagnostic Use

    Rat Anti-TERF1 Monoclonal Antibody, Clone 683D (CABT-RM172) This Product Is for Research Use Only and Is Not Intended for Diagnostic Use

    Rat Anti-TERF1 monoclonal antibody, clone 683D (CABT-RM172) This product is for research use only and is not intended for diagnostic use. PRODUCT INFORMATION Specificity Specifically detects murine Telomeric repeat-binding factor 1 (TRF1). Target TERF1 Immunogen His-tagged full-length recombinant mouse Telomeric repeat-binding factor 1 (TRF1). Isotype IgG1, κ Source/Host Rat Species Reactivity Mouse Clone 683D Purification Protein G purified Conjugate unconjugated Applications FC, ICC, IF, WB Molecular Weight ~51 kDa observed; 48.22 kDa calculated. Uncharacterized bands may be observed in some lysate(s). Format Liquid Size 100 μg, 25 μg Buffer 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl Preservative 0.05% sodium azide Storage Stable for 1 year at 2-8°C from date of receipt. Warnings Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. 45-1 Ramsey Road, Shirley, NY 11967, USA Email: [email protected] Tel: 1-631-624-4882 Fax: 1-631-938-8221 1 © Creative Diagnostics All Rights Reserved BACKGROUND Introduction Telomeric repeat-binding factor 1 is encoded by the Terf1 gene in murine species. TRF1 is a component of the shelterin complex that is involved in the regulation of telomere length and protection. It binds to telomeric DNA as a homodimer and protects telomeres.
  • Original Article Association of AXIN2 and MMP7 Polymorphisms with Non-Small Cell Lung Cancer in Chinese Han Population

    Original Article Association of AXIN2 and MMP7 Polymorphisms with Non-Small Cell Lung Cancer in Chinese Han Population

    Int J Clin Exp Pathol 2016;9(2):2253-2258 www.ijcep.com /ISSN:1936-2625/IJCEP0009888 Original Article Association of AXIN2 and MMP7 polymorphisms with non-small cell lung cancer in Chinese Han population Shuguang Han, Lei Lv, Xinhua Wang, Xun Wang, Hongqing Zhao Department of Respiratory Medicine, Second People’s Hospital of Wuxi, Wuxi, Jiangsu, China Received May 4, 2015; Accepted June 23, 2015; Epub February 1, 2016; Published February 15, 2016 Abstract: Objectives: This study aimed to explore the effect of AXIN2 and MMP7 polymorphisms on non-small cell lung cancer (NSCLC) susceptibility; in addition, the interaction between gene polymorphisms and environment was also displayed. Methods: The genotyping was conducted by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 102 patients with NSCLC and 120 healthy controls. Odds ratio (OR) and 95% con- fidence interval (CI) were calculated to assess the relevance strength of AXIN2 and MMP7 polymorphisms with NSCLC. The x² test was used to compare to the frequencies difference of genotypes and alleles in cases and controls and Hardy-Weinberg equilibrium (HWE) test. The haplotype and interaction analyses were performed by haploview and MDR software, respectively. Results: The genotype frequencies of all polymorphisms in the control group conformed to HWE. GG genotype frequency of AXIN2 rs2240307 polymorphism was significantly higher in cases than controls (P=0.041). Similarly, rs2240308 in AXIN2 gene was also increased the susceptibility to NSCLC remarkably (OR=2.412, 95% CI=1.025-5.674). What’s more, haplotype A-G-G in AXIN2 might play a protective role in NSCLC (OR=0.462, 95% CI=0.270-0.790).
  • Whole Proteome Analysis of Human Tankyrase Knockout Cells Reveals Targets of Tankyrase- Mediated Degradation

    Whole Proteome Analysis of Human Tankyrase Knockout Cells Reveals Targets of Tankyrase- Mediated Degradation

    ARTICLE DOI: 10.1038/s41467-017-02363-w OPEN Whole proteome analysis of human tankyrase knockout cells reveals targets of tankyrase- mediated degradation Amit Bhardwaj1, Yanling Yang2, Beatrix Ueberheide2 & Susan Smith1 Tankyrase 1 and 2 are poly(ADP-ribose) polymerases that function in pathways critical to cancer cell growth. Tankyrase-mediated PARylation marks protein targets for proteasomal 1234567890 degradation. Here, we generate human knockout cell lines to examine cell function and interrogate the proteome. We show that either tankyrase 1 or 2 is sufficient to maintain telomere length, but both are required to resolve telomere cohesion and maintain mitotic spindle integrity. Quantitative analysis of the proteome of tankyrase double knockout cells using isobaric tandem mass tags reveals targets of degradation, including antagonists of the Wnt/β-catenin signaling pathway (NKD1, NKD2, and HectD1) and three (Notch 1, 2, and 3) of the four Notch receptors. We show that tankyrases are required for Notch2 to exit the plasma membrane and enter the nucleus to activate transcription. Considering that Notch signaling is commonly activated in cancer, tankyrase inhibitors may have therapeutic potential in targeting this pathway. 1 Kimmel Center for Biology and Medicine at the Skirball Institute, Department of Pathology, New York University School of Medicine, New York, NY 10016, USA. 2 Proteomics Laboratory, Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA. Correspondence and requests for materials should be addressed to S.S. (email: [email protected]) NATURE COMMUNICATIONS | 8: 2214 | DOI: 10.1038/s41467-017-02363-w | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/s41467-017-02363-w ankyrases function in cellular pathways that are critical to function in human cells will provide insights into the clinical cancer cell growth including telomere cohesion and length utility of tankyrase inhibitors.
  • Mayer-Rokitansky-Küster- Hauser Syndrome

    Mayer-Rokitansky-Küster- Hauser Syndrome

    Morcel et al. Orphanet Journal of Rare Diseases 2011, 6:9 http://www.ojrd.com/content/6/1/9 RESEARCH Open Access Utero-vaginal aplasia (Mayer-Rokitansky-Küster- Hauser syndrome) associated with deletions in known DiGeorge or DiGeorge-like loci Karine Morcel1,2*†, Tanguy Watrin1†, Laurent Pasquier1,3, Lucie Rochard1, Cédric Le Caignec4,5, Christèle Dubourg1,6, Philippe Loget7, Bernard-Jean Paniel8, Sylvie Odent1,3, Véronique David1,6, Isabelle Pellerin1, Claude Bendavid1,6 and Daniel Guerrier1 Abstract Background: Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome is characterized by congenital aplasia of the uterus and the upper part of the vagina in women showing normal development of secondary sexual characteristics and a normal 46, XX karyotype. The uterovaginal aplasia is either isolated (type I) or more frequently associated with other malformations (type II or Müllerian Renal Cervico-thoracic Somite (MURCS) association), some of which belong to the malformation spectrum of DiGeorge phenotype (DGS). Its etiology remains poorly understood. Thus the phenotypic manifestations of MRKH and DGS overlap suggesting a possible genetic link. This would potentially have clinical consequences. Methods: We searched DiGeorge critical chromosomal regions for chromosomal anomalies in a cohort of 57 subjects with uterovaginal aplasia (55 women and 2 aborted fetuses). For this candidate locus approach, we used a multiplex ligation-dependent probe amplification (MLPA) assay based on a kit designed for investigation of the chromosomal regions known to be involved in DGS. The deletions detected were validated by Duplex PCR/liquid chromatography (DP/LC) and/or array-CGH analysis. Results: We found deletions in four probands within the four chromosomal loci 4q34-qter, 8p23.1, 10p14 and 22q11.2 implicated in almost all cases of DGS syndrome.
  • Crystal Structure of a Tankyrase-Axin Complex and Its Implications for Axin Turnover and Tankyrase Substrate Recruitment

    Crystal Structure of a Tankyrase-Axin Complex and Its Implications for Axin Turnover and Tankyrase Substrate Recruitment

    Crystal structure of a Tankyrase-Axin complex and its implications for Axin turnover and Tankyrase substrate recruitment Seamus Morronea,1, Zhihong Chenga,1, Randall T. Moonb, Feng Congc, and Wenqing Xua,2 aDepartment of Biological Structure, University of Washington School of Medicine, Seattle, WA 98195; bDepartment of Pharmacology, Howard Hughes Medical Institute, and Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, WA 98195; and cNovartis Institutes for Biomedical Research, Cambridge, MA 02139 Edited by* Stephen C. Harrison, Children's Hospital, Harvard Medical School, and Howard Hughes Medical Institute, Boston, MA, and approved December8, 2011 (received for review October 9, 2011) Axin is a tumor suppressor and a key negative regulator of the Ubiquitination of Axin, which leads to its subsequent turnover, Wnt/β-catenin signaling pathway. Axin turnover is controlled by its requires its poly-ADP-ribosylation (PARylation) (11). PARylation poly-ADP-ribosylation catalyzed by tankyrase (TNKS), which re- of proteins is catalyzed by a family of poly-ADP-ribose poly- quires the direct interaction of Axin with TNKS. This interaction merases (PARPs), with 18 known members in humans, which re- is thus an attractive drug target for treating cancers, brain injuries, gulate many aspects of biology including genomic stability, cell and other diseases where β-catenin is involved. Here we report the cycle, and energy metabolism (15, 16). Axin PARylation is speci- crystal structure of a mouse TNKS1 fragment containing ankyrin- fically catalyzed by tankyrases 1 and 2 (a.k.a. TNKS1/PARP5a repeat clusters 2 and 3 (ARC2-3) in a complex with the TNKS-bind- and TNKS2/PARP5b, respectively).
  • Β-Catenin-Mediated Wnt Signal Transduction Proceeds Through an Endocytosis-Independent Mechanism

    Β-Catenin-Mediated Wnt Signal Transduction Proceeds Through an Endocytosis-Independent Mechanism

    bioRxiv preprint doi: https://doi.org/10.1101/2020.02.13.948380; this version posted February 20, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. β-catenin-Mediated Wnt Signal Transduction Proceeds Through an Endocytosis-Independent Mechanism Ellen Youngsoo Rim1, , Leigh Katherine Kinney1, and Roel Nusse1, 1Howard Hughes Medical Institute, Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA The Wnt pathway is a key intercellular signaling cascade that by GSK3β is inhibited. This leads to β-catenin accumulation regulates development, tissue homeostasis, and regeneration. in the cytoplasm and concomitant translocation into the nu- However, gaps remain in our understanding of the molecular cleus, where it can induce transcription of target genes. The events that take place between ligand-receptor binding and tar- importance of β-catenin stabilization in Wnt signal transduc- get gene transcription. Here we used a novel tool for quanti- tion has been demonstrated in many in vivo and in vitro con- tative, real-time assessment of endogenous pathway activation, texts (8, 9). However, immediate molecular responses to the measured in single cells, to answer an unresolved question in the ligand-receptor interaction and how they elicit accumulation field – whether receptor endocytosis is required for Wnt signal transduction. We combined knockdown or knockout of essential of β-catenin are not fully elucidated. components of Clathrin-mediated endocytosis with quantitative One point of uncertainty is whether receptor endocyto- assessment of Wnt signal transduction in mouse embryonic stem sis following Wnt binding is required for signal transduc- cells (mESCs).
  • Discovery of a Novel Triazolopyridine Derivative As a Tankyrase Inhibitor

    Discovery of a Novel Triazolopyridine Derivative As a Tankyrase Inhibitor

    International Journal of Molecular Sciences Article Discovery of a Novel Triazolopyridine Derivative as a Tankyrase Inhibitor Hwani Ryu 1, Ky-Youb Nam 2, Hyo Jeong Kim 1, Jie-Young Song 1 , Sang-Gu Hwang 1 , Jae Sung Kim 1 , Joon Kim 3,* and Jiyeon Ahn 1,* 1 Division of Radiation Biomedical Research, Korea Institute of Radiological & Medical Sciences, Seoul 01812, Korea; [email protected] (H.R.); [email protected] (H.J.K.); [email protected] (J.-Y.S.); [email protected] (S.-G.H.); [email protected] (J.S.K.) 2 Department of Research Center, Pharos I&BT Co., Ltd., Anyang 14059, Korea; [email protected] 3 Laboratory of Biochemistry, Division of Life Sciences, Korea University, Seoul 02841, Korea * Correspondence: [email protected] (J.K.); [email protected] (J.A.); Tel.: +82-2-970-1311 (J.A.) Abstract: More than 80% of colorectal cancer patients have adenomatous polyposis coli (APC) mutations, which induce abnormal WNT/β-catenin activation. Tankyrase (TNKS) mediates the release of active β-catenin, which occurs regardless of the ligand that translocates into the nucleus by AXIN degradation via the ubiquitin-proteasome pathway. Therefore, TNKS inhibition has emerged as an attractive strategy for cancer therapy. In this study, we identified pyridine derivatives by evaluating in vitro TNKS enzyme activity and investigated N-([1,2,4]triazolo[4,3-a]pyridin-3-yl)-1-(2- cyanophenyl)piperidine-4-carboxamide (TI-12403) as a novel TNKS inhibitor. TI-12403 stabilized β β AXIN2, reduced active -catenin, and downregulated -catenin target genes in COLO320DM and DLD-1 cells.
  • Mutational Analysis of AXIN2, MSX1, and PAX9 in Two Mexican Oligodontia Families

    Mutational Analysis of AXIN2, MSX1, and PAX9 in Two Mexican Oligodontia Families

    Mutational analysis of AXIN2, MSX1, and PAX9 in two Mexican oligodontia families Y.D. Mu1,2, Z. Xu1, C.I. Contreras1, J.S. McDaniel1, K.J. Donly1 and S. Chen1 1Department of Developmental Dentistry, Dental School, University of Texas Health Science Center, San Antonio, Texas, USA 2Stomotology Department, Sichuan Provincial People’s Hospital, Chengdu, Sichuan, China Corresponding author: S. Chen E-mail: [email protected] Genet. Mol. Res. 12 (4): 4446-4458 (2013) Received July 4, 2012 Accepted May 15, 2013 Published October 10, 2013 DOI http://dx.doi.org/10.4238/2013.October.10.10 ABSTRACT. The genes for axin inhibition protein 2 (AXIN2), msh homeobox 1 (MSX1), and paired box gene 9 (PAX9) are involved in tooth root formation and tooth development. Mutations of the AXIN2, MSX1, and PAX9 genes are associated with non-syndromic oligodontia. In this study, we investigated phenotype and AXIN2, MSX1, and PAX9 gene variations in two Mexican families with non-syndromic oligodontia. Individuals from two families underwent clinical examinations, including an intra-oral examination and panoramic radiograph. Retrospective data were reviewed, and peripheral blood samples were collected. The exons and exon-intronic boundaries of the AXIN2, MSX1, and PAX9 genes were sequenced and analyzed. Protein and messenger RNA structures were predicted using bioinformative software programs. Clinical and oral examinations revealed isolated non-syndromic oligodontia in the two Mexican families. The average number of missing teeth was 12. The sequence analysis of exons and exon-intronic regions of AXIN2, MSX1, and PAX9 revealed 11 single- nucleotide polymorphisms (SNPs), including seven in AXIN2, two Genetics and Molecular Research 12 (4): 4446-4458 (2013) ©FUNPEC-RP www.funpecrp.com.br AXIN2, MSX1, and PAX9 SNPs and oligodontia 4447 in MSX1, and three in PAX9.
  • Novel Missense Mutations in the AXIN2 Gene Associated with Non

    Novel Missense Mutations in the AXIN2 Gene Associated with Non

    a r c h i v e s o f o r a l b i o l o g y 5 9 ( 2 0 1 4 ) 3 4 9 – 3 5 3 Available online at www.sciencedirect.com ScienceDirect journal homepage: http://www.elsevier.com/locate/aob Novel missense mutations in the AXIN2 gene associated with non-syndromic oligodontia a,1 a,1 b a Singwai Wong , Haochen Liu , Baojing Bai , Huaiguang Chang , c,d e a, a, Hongshan Zhao , Yixiang Wang , Dong Han **, Hailan Feng * a Department of Prosthodontics, Peking University School and Hospital of Stomatology, Beijing, China b Department of Prosthodontics, Beijing Stomatology Hospital, Beijing, China c Department of Medical Genetics, Peking University Health Science Center, Beijing, China d Peking University Center for Human Disease Genomics, Peking University Health Science Center, Beijing, China e Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, China a r t i c l e i n f o a b s t r a c t Article history: Objective: Oligodontia, which is the congenital absence of six or more permanent teeth Accepted 23 December 2013 excluding third molars, may contribute to masticatory dysfunction, speech alteration, aesthetic problems and malocclusion. To date, mutations in EDA, AXIN2, MSX1, PAX9, Keywords: WNT10A, EDAR, EDARADD, NEMO and KRT 17 are known to associate with non-syndromic oligodontia. The aim of the study was to search for AXIN2 mutations in 96 patients with non- Non-syndromic oligodontia syndromic oligodontia. Mutation screening AXIN2 Design: We performed mutation analysis of 10 exons of the AXIN2 gene in 96 patients with isolated non-syndromic oligodontia.
  • Hypodontia As a Risk Marker for Epithelial Ovarian Cancer a Case-Controlled Study

    Hypodontia As a Risk Marker for Epithelial Ovarian Cancer a Case-Controlled Study

    RESEARCH Hypodontia as a risk marker for epithelial ovarian cancer A case-controlled study Leigh A. Chalothorn, DMD, MS; Cynthia S. Beeman, DDS, PhD; Jeffrey L. Ebersole, PhD; G. Thomas Kluemper, DMD, MS; E. Preston Hicks, DDS, MS, MSD; Richard J. Kryscio, PhD; Christopher P. DeSimone, MD; Susan C. Modesitt, MD ecent advances in genetic research have allowed for A D A a greater understanding ABSTRACT J 2 2 of the underlying causes C N of human disease, fur- Background. Genetic mutations that result in O O N I thering the exploration and insight hypodontia also may be associated with abnormali- T T R I A N U C into possible genetic associations ties in other parts of the body. The authors conducted I U A N G E D among seemingly unrelated condi- a study to establish the prevalence rates of hypodontia R 2 TI E tions. More than 300 genes are among subjects with epithelial ovarian cancer (EOC) and CL involved in odontogenesis, and control subjects to explore possible genetic associations between these mutations in several of these genes two phenotypes. have been linked with hypodontia.1,2 Methods. The authors recruited 50 subjects with EOC and 100 control The genes that control the develop- subjects who did not have EOC. The authors performed a dental exami- ment of teeth also have important nation on each subject to detect hypodontia, and they reviewed pertinent functions in other organs and body radiographs and dental histories. They also recorded any family history systems. Therefore, it is plausible to of cancer and hypodontia.
  • Structural Basis and Sequence Rules for Substrate Recognition by Tankyrase Explain the Basis for Cherubism Disease

    Structural Basis and Sequence Rules for Substrate Recognition by Tankyrase Explain the Basis for Cherubism Disease

    Structural Basis and Sequence Rules for Substrate Recognition by Tankyrase Explain the Basis for Cherubism Disease Sebastian Guettler,1,2 Jose LaRose,3 Evangelia Petsalaki,1,2 Gerald Gish,1 Andy Scotter,3 Tony Pawson,1,2,* Robert Rottapel,3,4,* and Frank Sicheri1,2,* 1Centre for Systems Biology, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada 2Department of Molecular Genetics, University of Toronto, 1 Kings College Circle, Toronto, Ontario M5S 1A8, Canada 3Ontario Cancer Institute and the Campbell Family Cancer Research Institute, 101 College Street, Room 8-703, Toronto Medical Discovery Tower, University of Toronto, Toronto, Ontario M5G 1L7, Canada 4Division of Rheumatology, Department of Medicine, Saint Michael’s Hospital, 30 Bond Street, Toronto, Ontario M5B 1W8, Canada *Correspondence: [email protected] (T.P.), [email protected] (R.R.), [email protected] (F.S.) DOI 10.1016/j.cell.2011.10.046 SUMMARY asparagine, arginine, lysine, cysteine, phosphoserine, and diph- thamide residues (reviewed in Hottiger et al., 2010). As a large The poly(ADP-ribose)polymerases Tankyrase 1/2 posttranslational modification of substantial negative charge, (TNKS/TNKS2) catalyze the covalent linkage of protein poly(ADP-ribosyl)ation (PARsylation) can influence pro- ADP-ribose polymer chains onto target proteins, tein fate through several mechanisms, including a direct effect regulating their ubiquitylation, stability, and function. on protein activity, recruitment of binding partners
  • Wnt/Β-Catenin Signaling Via Axin2 Is Required for Myogenesis And

    Wnt/Β-Catenin Signaling Via Axin2 Is Required for Myogenesis And

    © 2016. Published by The Company of Biologists Ltd | Development (2016) 143, 3128-3142 doi:10.1242/dev.139907 RESEARCH ARTICLE Wnt/β-catenin signaling via Axin2 is required for myogenesis and, together with YAP/Taz and Tead1, active in IIa/IIx muscle fibers Danyil Huraskin1,‡, Nane Eiber1,‡, Martin Reichel2,*, Laura M. Zidek3, Bojana Kravic1, Dominic Bernkopf2, Julia von Maltzahn3,Jürgen Behrens2 and Said Hashemolhosseini1,¶ ABSTRACT are expressed in the dorsal ectoderm, and Wnt11 is expressed in the Canonical Wnt/β-catenin signaling plays an important role in epaxial dermomyotome. In adult muscle, Wnt proteins are expressed myogenic differentiation, but its physiological role in muscle fibers upon regeneration (Polesskaya et al., 2003; von Maltzahn et al., remains elusive. Here, we studied activation of Wnt/β-catenin 2012). Non-canonical Wnt signaling through Wnt7a regulates signaling in adult muscle fibers and muscle stem cells in an Axin2 satellite cell number and the size of skeletal muscle (Le Grand et al., reporter mouse. Axin2 is a negative regulator and a target of Wnt/β- 2009; von Maltzahn et al., 2013). Generally, non-canonical Wnt catenin signaling. In adult muscle fibers, Wnt/β-catenin signaling is signaling is required for skeletal muscle development, while only detectable in a subset of fast fibers that have a significantly canonical Wnt signaling, especially via Wnt3a, has been smaller diameter than other fast fibers. In the same fibers, demonstrated to lead to increased fibrosis (Brack et al., 2007). immunofluorescence staining for YAP/Taz and Tead1 was In canonical Wnt signaling the Wnt binds to the Fzd and Lrp5/6 detected.