Number 2 February 2015

Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Volume 19 - Number 2 February 2015

The PDF version of the Atlas of Genetics and Cytogenetics in Oncology and Haematology is a reissue of the original articles published in collaboration with the Institute for Scientific and Technical Information (INstitut de l’Information Scientifique et Technique - INIST) of the French National Center for Scientific Research (CNRS) on its electronic publishing platform I-Revues. Online and PDF versions of the Atlas of Genetics and Cytogenetics in Oncology and Haematology are hosted by INIST-CNRS. Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Scope

The Atlas of Genetics and Cytogenetics in Oncology and Haematology is a peer reviewed on-line journal in open access, devoted to genes, cytogenetics, and clinical entities in cancer, and cancer-prone diseases. It presents structured review articles (“cards”) on genes, leukaemias, solid tumours, cancer-prone diseases, and also more traditional review articles (“deep insights”) on the above subjects and on surrounding topics. It also present case reports in hematology and educational items in the various related topics for students in Medicine and in Sciences.

Editorial correspondance

Jean-Loup Huret Genetics, Department of Medical Information, University Hospital F-86021 Poitiers, France tel +33 5 49 44 45 46 [email protected] or [email protected]

Staff Mohammad Ahmad, Mélanie Arsaban, Marie-Christine Jacquemot-Perbal, Vanessa Le Berre, Anne Malo, Carol Moreau, Catherine Morel-Pair, Laurent Rassinoux, Alain Zasadzinski. Philippe Dessen is the Database Director (Gustave Roussy Institute – Villejuif – France).

The Atlas of Genetics and Cytogenetics in Oncology and Haematology (ISSN 1768-3262) is published 12 times a year by ARMGHM, a non profit organisation, and by the INstitute for Scientific and Technical Information of the French National Center for Scientific Research (INIST-CNRS) since 2008.

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Editor

Jean-Loup Huret (Poitiers, France) Editorial Board

Sreeparna Banerjee (Ankara, Turkey) Solid Tumours Section Alessandro Beghini (Milan, Italy) Genes Section Anne von Bergh (Rotterdam, The Netherlands) Genes / Leukaemia Sections Judith Bovée (Leiden, The Netherlands) Solid Tumours Section Vasantha Brito-Babapulle (London, UK) Leukaemia Section Charles Buys (Groningen, The Netherlands) Deep Insights Section Antonio Cuneo (Ferrara, Italy) Leukaemia Section Paola Dal Cin (Boston, Massachussetts) Genes / Solid Tumours Section Brigitte Debuire (Villejuif, France) Deep Insights Section Marc De Braekeleer (Brest, France) Genes / Leukaemia Sections François Desangles (Paris, France) Leukaemia / Solid Tumours Sections Enric Domingo-Villanueva (London, UK) Solid Tumours Section Ayse Erson (Ankara, Turkey) Solid Tumours Section Richard Gatti (Los Angeles, California) Cancer-Prone Diseases / Deep Insights Sections Ad Geurts van Kessel (Nijmegen, The Netherlands) Cancer-Prone Diseases Section Oskar Haas (Vienna, Austria) Genes / Leukaemia Sections Anne Hagemeijer (Leuven, Belgium) Deep Insights Section Nyla Heerema (Colombus, Ohio) Leukaemia Section Jim Heighway (Liverpool, UK) Genes / Deep Insights Sections Sakari Knuutila (Helsinki, Finland) Deep Insights Section Lidia Larizza (Milano, Italy) Solid Tumours Section Lisa Lee-Jones (Newcastle, UK) Solid Tumours Section Edmond Ma (Hong Kong, China) Leukaemia Section Roderick McLeod (Braunschweig, Germany) Deep Insights / Education Sections Cristina Mecucci (Perugia, Italy) Genes / Leukaemia Sections Fredrik Mertens (Lund, Sweden) Solid Tumours Section Konstantin Miller (Hannover, Germany) Education Section Felix Mitelman (Lund, Sweden) Deep Insights Section Hossain Mossafa (Cergy Pontoise, France) Leukaemia Section Stefan Nagel (Braunschweig, Germany) Deep Insights / Education Sections Florence Pedeutour (Nice, France) Genes / Solid Tumours Sections Elizabeth Petty (Ann Harbor, Michigan) Deep Insights Section Susana Raimondi (Memphis, Tennesse) Genes / Leukaemia Section Mariano Rocchi (Bari, Italy) Genes Section Alain Sarasin (Villejuif, France) Cancer-Prone Diseases Section Albert Schinzel (Schwerzenbach, Switzerland) Education Section Clelia Storlazzi (Bari, Italy) Genes Section Sabine Strehl (Vienna, Austria) Genes / Leukaemia Sections Nancy Uhrhammer (Clermont Ferrand, France) Genes / Cancer-Prone Diseases Sections Dan Van Dyke (Rochester, Minnesota) Education Section Roberta Vanni (Montserrato, Italy) Solid Tumours Section Franck Viguié (Paris, France) Leukaemia Section José Luis Vizmanos (Pamplona, Spain) Leukaemia Section Thomas Wan (Hong Kong, China) Genes / Leukaemia Sections Adriana Zamecnikova (Kuwait) Leukaemia Section

Atlas Genet Cytogenet Oncol Haematol. 2015 19(2) Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Volume 19, Number 2, February 2015

Table of contents

Gene Section

BIN1 (bridging integrator 1) 78 Sunil Thomas, Mee Young Chang, George C Prendergast CDH3 (cadherin 3, type 1, P-cadherin (placental)) 85 André Filipe Vieira, Ana Sofia Ribeiro, Joana Paredes CELF2 (CUGBP, Elav-like family member 2) 93 Satish Ramalingam, Shrikant Anant CXCL17 (chemokine (C-X-C motif) ligand 17) 97 Aya Matsui, Takashi Murakami LRIG3 (leucine-rich repeats and immunoglobulin-like domains 3) 102 Dongsheng Guo, Feng Mao, Ting Lei MKI67 (marker of proliferation Ki-67) 105 Konstantinos Ntzeros, Philip Stanier, Dimitrios Mazis, Neoklis Kritikos, Meletios Rozis, Eustathios Anesidis, Chrisoula Antoniou, Michael Stamatakos OSGIN1 (oxidative stress induced growth inhibitor 1) 117 Jing Hu, Yanming Wang PTPRA (protein tyrosine phosphatase, receptor type, A) 121 Jian Huang, Xueping Lai, Xinmin Zheng STMN1 (stathmin 1) 126 João Agostinho Machado-Neto, Fabiola Traina MTA3 (metastasis associated 1 family, member 3 ) 137 Ansgar Brüning, Ioannis Mylonas

Leukaemia Section t(10;11)(q22;q23) KMT2A/TET1 141 Antoine Ittel t(2;9)(p23;q33) TRAF1/ALK 145 Xiaoming Xing, Andrew L Feldman t(9;15)(p13;q24) PAX5/GOLGA6A 148 Jean-Loup Huret

Solid Tumour Section

Bone: Epithelioid hemangioendothelioma 150 Andreas F Mavrogenis, Andrea Angelini, Costantino Errani, Pietro Ruggieri

Atlas Genet Cytogenet Oncol Haematol. 2015 19(2)

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in Oncology and Haematology

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Deep Insight Section

Insights into structure and function of human PWWP domains 155 Su Qin, Jinrong Min

Atlas Genet Cytogenet Oncol Haematol. 2015 19(2)

Atlas of Genetics and Cytogenetics

in Oncology and Haematology

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Gene Section Review

BIN1 (bridging integrator 1) Sunil Thomas, Mee Young Chang, George C Prendergast Lankenau Institute for Medical Research, Wynnewood, Pennsylvania 19096, USA (ST, MYC, GCP)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/BIN1ID794ch2q14.html DOI: 10.4267/2042/56290 This article is an update of : Chang MY, Prendergast GC. BIN1 (bridging integrator 1). Atlas Genet Cytogenet Oncol Haematol 2009;13(8):543-548.

localized to a syntenic locus on mouse chromosome Abstract 8 (Wechsler-Reya et al., 1997). The BIN1 gene encodes a set of BAR adapter Transcription proteins generated by alternate RNA splicing which function in membrane and actin dynamics, cell The Bin1 promoter is rich in CpG methylation polarity and stress signaling. In cancer cells, BIN1 residues and transcription of the gene produces functions as a tumor suppressor gene and it is more than 10 alternative transcripts that are from commonly silenced or misspliced during malignant 2075 to 2637 bp mRNA in size: isoform 1 (2637 progression. Genetic investigations in fission yeast, bp), isoform 2 (2508 bp), isoform 3 (2376 bp), flies, mice and human cells suggest that BIN1 isoform 4 (2333 bp), isoform 5 (2412 bp), isoform adapter proteins exert a 'moonlighting' function in 6 (2289 bp), isoform 7 (2283 bp), isoform 8 (2210) the nucleus, integrating cell polarity signals bp, isoform 9 (2165bp), and isoform 10 (2075 bp). generated by actin and vesicle dynamics with Isoforms 9 and 10 are ubiquitous in expression. central regulators of cell cycle control, apoptosis, Isoform 8 is expressed specifically in skeletal and immune surveillance (adapted from Prendergast muscle. et al., 2009). Isoforms 1-7 are expressed predominantly in the central nervous system. An aberrant isoform has Keywords been reported to be expressed specifically in tumor Tumor suppression, apoptosis, actin-membrane cells (Wechsler-Reya et al., 1997). dynamics, vesicle trafficking, immune surveillance Pseudogene Identity None reported. Other names: AMPH-II, AMPH2, AMPHL, ALP, DKFZp547F068, MGC10367, SH3P9 Protein HGNC (Hugo): BIN1 Description Location: 2q14.3 Bin1 contains N-terminal BAR (Bin1/Amphihysin/Rvs) domain with predicted DNA/RNA coiled-coil structure and a C-terminal SH3 domain (Sakamuro et al., 1996). Bin1 encodes proteins of Description 409 to 593 amino acids; isoform 1 (593 aa), isoform The human BIN1 gene is encoded by at least 16 2 (550 aa), isoform 3 (506 aa), isoform 4 (497 aa), exons spanning at least 59258 bps at chromosome isoform 5 (518 aa), isoform 6 (482 aa), isoform 7 2q14-2q21 (nucleotides 127522078-127581334). (475 aa), isoform 8 (454 aa), isoform 9 (439 aa), The murine Bin1 gene is similarly sized but and isoform 10 (409 aa).

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At least 10 alternate protein isoforms of Bin1 are expressed in different tissues. Isoforms 9 and 10 are ubiquitous. Isoform 8 is muscle-specific. Isoforms 1-7 are expressed predominantly in the central nervous system. Two tumor-specific isoforms include an exon termed 12A that is normally spliced into Bin1 mRNA only with other exons expressed in the central nervous system. These tumor-specific isoforms occur commonly in cancer and they represent loss of function with regard to tumor suppression activity and nuclear localization capability. BAR, BAR domain; SH3, SH3 domain; MBD, Myc binding domain; CLAP, clathrin- associated protein binding region; PI, phosphoinositide binding region. Exons are numbered by reference to Wechsler-Reya et al. (1997).

Isoform 10 is the smallest and isoform 1 is the membrane localized in gastrointestinal cells largest in size. Also, Bin1 has predicted molecular (DuHadaway et al., 2003). In cardiac muscles Bin1 weight of 45432 to 64568 Da; isoform 1 (64568 generate T-tubules and also designates T-tubules as Da), isoform 2 (59806 Da), isoform 3 (55044 Da), the appropriate site for delivery of L-type calcium isoform 4 (54817 Da), isoform 5 (56368 Da), channels (Hong et al., 2010). isoform 6 (52889 Da), isoform 7 (51606 Da), Function isoform 8 (50054 Da), isoform 9 (48127 Da), and isoform 10 (45432 Da). Isoforms 9 and 10 are Bin1 encodes members of the BAR ubiquitous in expression. Isoform 8 is expressed (Bin/Amphiphysin/Rvs) adapter family which have specifically in skeletal muscle. Isoforms 1-7 are been implicated in membrane dynamics, such as expressed predominantly in the central nervous vesicle fusion and trafficking, specialized system. These 10 different splice isoforms differ membrane organization, actin organization, cell widely in subcellular localization, tissue polarity, stress signaling, transcription, distribution, and ascribed functions, with isoforms immunomodulation and tumor suppression. BAR 1-7 predominantly cytosolic but isoforms 8-10 adapter proteins are now recognized to be part of a found in both the nucleus and/or cytosol of certain larger superfamily of structurally related proteins cell types (Sakamuro et al., 1996; Muller et al., that includes the so-called F-BAR and I-BAR 2003). Recent studies demonstrated that the coiled- adapter proteins (Ren et al., 2006; Prendergast et coil BIN1 BAR peptide encodes a novel BIN1 MID al., 2009). domain, through which BIN1 acts as a MYC- Membrane binding and tubulation: The Bin1 independent cancer suppressor (Lundgaard et al., BAR domain can mediate binding and tubulation of 2011). curved membranes (Lee et al., 2002; Wu et al., 2014). Crystal structures of the BAR domains from Expression human BIN1 and its fruit fly homolog reveal a Bin1 is widely expressed (Wechsler-Reya et al., dimeric banana-shaped 6-alpha-helix bundle that 1997; Chapuis et al., 2013). Patterns of isoform can nestle against the charged head groups on a expression are noted above in the diagram legend. curved lipid bilayer. Structural studies implicate specific alpha-helices in tubulation activity. Localisation Biochemical analyses implicate Bin1 in vesicle Bin1 is localized both in nuclear and cytosolic in fission and fusion processes, with the SH3 domain the cerebral cortex and cerebellum of brain. Bin1 is providing an essential contribution to these localized mainly in nuclear in bone marrow cells processes through the recruitment of dynamins whereas it is localized mainly in cytosolic in (Ren et al., 2006). peripheral lymphoid cells. Bin1 is nuclear or Vesicle trafficking: Bin1 is implicated in nucleocytosolic in basal cells of skin, breast, or endocytosis and intracellular endosome traffic prostate, whereas it is cytosolic or plasma through interactions with Rab5 guanine nucleotide

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 79 BIN1 (bridging integrator 1) Thomas S, et al.

exchange factors (Rab GEFs) and the sorting nexin controls the paracellular pathway of transcellular protein Snx4. Complexes of neuronal Amph-I with ion transport regulated by cellular tight junctions neuron-specific isoforms of Bin1 (Amph-II) have (Chang et al., 2012). been implicated in synaptic vesicle recycling in the Apoptosis and Senescence: Bin1 is crucial for the brain. Genetic studies of the Bin1 homolog in function of default pathways of classical apoptosis budding yeast indicate an essential role in or senescence triggered by the Myc or Raf endocytosis, however, this role appears to be non- oncogenes in primary cells (Prendergast et al., essential for homologs in fission yeast, fruit flies, 2009). In human tumor cells, enforced expression and mice (Leprince et al., 2003). of Bin1 triggers a non-classical program of cell Cell polarity: Genetic analyses of the Bin1 death that is caspase independent and associated homologs in yeast and fruit flies suggest a with activation of serine proteases (Elliott et al., integrative function in cell polarity, possibly 2000). mediated by effects on actin organization and Tumor suppression: Attenuation of Bin1 function vesicle trafficking. In budding yeast, the Bin1 by silencing or missplicing is a frequent event in homolog RVS167 lies at a central nodal point for multiple human cancers including breast, prostate, integrating cell polarity signaling (Balguerie et al., skin, lung, and colon cancers (Sakamuro et al., 1999). Genetic ablation of the Bin1 homolog in 1996; Ge et al., 2000b). In breast cancer, attenuated fruit flies causes mislocalization of the cell polarity expression of Bin1 is associated with increased complex Dlg/Scr/Lgl, normally localized to the metastasis and poor clinical outcome (Chang et al., tight junction, that is implicated in epithelial 2007b). In human tumor cells, ectopic expression of polarity and suppression of tumor-like growths in ubiquitous or muscle Bin1 isoforms causes growth flies (Humbert et al., 2003). arrest or caspase-independent cell death Transcription: Ubiquitous and muscle-specific (Prendergast et al., 2009). A Bin1 missplicing event isoforms of Bin1 that can localize to the nucleus that occurs frequently in human cancers is sufficient can bind to c-Myc and suppress its transcriptional to extinguish these activities. In primary rodent transactivation activity (Elliott et al., 1999). cells, Bin1 inhibits oncogenic co-transformation by Tethering the BAR domain of Bin1 to DNA is Myc, adenovirus E1A, or mutant p53 but not SV40 sufficient to repress transcription. Genetic studies in T antigen (Elliott et al., 1999; Elliott et al., 2000). fission yeast demonstrate that the functional Mouse genetic studies establish that loss of Bin1 homolog hob1+ is essential to silence transcription causes lung and liver cancers during aging (Chang of heterochromatin at telomeric and centromeric et al., 2007a). In mice where breast or colon tumors chromosomal loci by supporting a Rad6-Set1 are initiated by carcinogen treatment, Bin1 deletion pathway of transcriptional repression (Ramalingam causes progression to more aggressive malignant and Prendergast, 2007). states (Chang et al., 2007b). Oncogenically Muscle function: Mutations of the human BIN1 transformed cells lacking Bin1 exhibit reduced gene are associated with centronuclear myopathy, a susceptibility to apoptosis and increased disorder marked by severe muscle weakness (Nicot proliferation, invasion, immune escape, and tumor et al., 2007; Wu et al., 2014). In skeletal muscle, formation (Muller et al., 2004, Muller et al., 2005). Bin1 localizes to T tubules where it appears to Activation of metalloproteinase MMP9 and support ion flux (Lee et al., 2002; Butler et al., immunosuppressive enzyme indoleamine 2,3- 1997). In vitro studies of terminal muscle dioxygenase (IDO) have been implicated differentiation implicate Bin1 in myoblast cell cycle respectively in invasion and immune escape caused arrest and fusion during tubule formation by Bin1 loss (Chang et al., 2007b). Cisplatin is the (Wechsler-Reya et al., 1998). most important and efficacious chemotherapeutic Cardiac function: Mouse genetic studies indicate agent for the treatment of advanced gastric cancer. that Bin1 is required for cardiac development The oncoprotein c-Myc suppresses bridging (Hong et al., 2010). Bin1 levels decreases in failing integrator 1 (BIN1), thereby releasing poly(ADP- hearts and low level of plasma Bin1 correlates with ribose)polymerase 1, which results in increased heart failure and predicts arrhythmia in patients DNA repair activity and allows cancer cells to with arrhythmogenic right ventricular acquire cisplatin resistance (Tanida et al., 2012). cardiomyopathy (Hong et al., 2012). Null phenotype in mouse: Bin1 knockout mice are Cognition and Memory: Bin1 is one of the perinatal lethal owing to myocardial hypertrophy candidate genes involved in Alzheimer's disease. where myofibrils of ventricular cardiomyocytes are Bin1 is the most important risk locus for late onset severely disorganized (Muller et al., 2003). Genetic Alzheimer's disease. Bin1 affects AD risk primarily mosaic mice display increased susceptibility to by modulating tau pathology (Tan et al., 2013; inflammation, premalignant lesions in prostate and Kingwell, 2013; Chapius et al., 2013). pancreas, and formation of liver and lung Immunomodulation and Barrier Function: Bin1 carcinoma. Female mosaic mice exhibit increased is a genetic modifier of experimental colitis that fecundity during aging. Tissue-specific gene

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ablation in skin or breast facilitates carcinogenesis Mouse genetic studies indicate that Bin1 is non- (Chang et al., 2007a). essential for mammary gland development but that it is needed for the rapid kinetics of ductolobular Homology remodeling during pregnancy and weaning. The longest Bin1 alternate splice variant in human In mammary gland tumors initiated by the ras- brain exhibits 71% amino acid sequences similarity activating carcinogen 7,12-dimethylbenzanthracene and 55% amino acid sequence identity with (DMBA), Bin1 loss strongly accentuates the amphiphysin-I (amph-I) (Ren et al., 2006). Bin1 is formation of poorly differentiated tumors also closely related to the mammalian amphiphysin- characterized by low tubule formation, high mitotic like genes Bin2 and Bin3 (Routhier et al., 2003). indices, and high degree of nuclear pleomorphism. Genetic homologs of Bin1 that exist in budding and Bin1 loss facilitates tumor progression at several fission yeast (RVS167 and hob1+) and in fruit flies intrinsic levels, including increased proliferation, (amphiphysin) are well-characterized (Ren et al., survival, and motility of mouse mammary epithelial 2006; Prendergast et al., 2009). cells (MMECs) established from DMBA-induced Mutations tumors (Chang et al., 2007b). Lung cancer Note Oncogenesis Epigenetics: Attenuated expression or missplicing Lung cancer is a leading cause of death from occurs in many cases of breast, prostate, lung, skin, cancer. brain and colon cancers. Expression analyses have Bin1 expression has been reported to be attenuated identified Bin1 missplicing as among the most significantly in cases of lung adenocarcinoma by common missplicing events occurring in cancer immunohistochemical analysis. (Chang et al., 2007). Mouse genetic studies demonstrate that Bin1 loss is Germinal associated with formation of lung adenocarcinoma during aging, indicating that Bin1 attenuation Germ-line mutations leading to nonsynonymous drives disease incidence (Chang et al., 2007a). alterations in Bin1 have been associated with the familial muscle weakness disorder centronuclear Colon cancer myopathy, a disorder characterized by abnormal Oncogenesis centralization of nuclei in muscle fibers. Two Colorectal cancer is the third most common cancer missense alterations the BAR domain that have in the developed world. been identified as loss of function mutations for Bin1 expression has been reported to be attenuated membrane binding are K35N and D151N (Nicot et significantly in ~50% of cases of colon cancer by al., 2007). Another mutation that has been Immunohistochemical analysis. identified, K575X, generates a prematurely Mouse genetic studies indicate that, in colon tumors terminated Bin1 protein implicated in loss of initiated by treatment with the carcinogen function (Nicot et al., 2007). dimethylhydrazine (DMH), Bin1 loss facilitates Somatic progression to more aggressive tumors with a higher multiplicity (Chang et al., 2007a). Loss of heterozygosity of Bin1 in cases of metastatic prostate cancer has been reported in the Skin cancer absence of mutation of the remaining allele. Oncogenesis Infrequent instances of gene deletions have been Skin cancer is the most common form of cancer. reported in breast tumor cell lines (Chang et al., Basal cell cancer and squamous cell cancer are 2007b; Kuznetsova et al., 2007). most common and treatable whereas melanoma is less common and deadlier. Studies of human Implicated in melanoma revealed that Bin1 is inappropriately Breast cancer expressed as tumor cell-specific isoforms that include exon 12A, which is alternately spliced into Oncogenesis isoforms found in the central nervous system but Breast cancer is the second leading cause of cancer not normally on its own in melanocytes or other death in women following lung cancer. Bin1 non-neuronal cells. This aberrant splicing event expression is attenuated significantly in >50% of abolishes the tumor suppressor functions of Bin1 cases of malignant breast cancer by based on the loss of its anti-oncogenic and immunohistochemical or RT-PCR analysis (Ge et programmed cell death inducing activities in al., 2000a). Reduced levels of Bin1 are correlated to oncogenically transformed cells and melanoma increased nodal metastasis and reduced survival in cells. Mouse genetic studies indicate that Bin1 loss low or middle grade carcinomas. facilitates skin carcinogenesis (Ge et al., 1999).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 81 BIN1 (bridging integrator 1) Thomas S, et al.

Prostate cancer were identified along one mutation causing a prematurely terminated Bin1 polypeptide (Nicot et Oncogenesis al., 2007). Prostate cancer is the leading cause of death in men older than 55 years of age. Loss of heterozygosity Chronic inflammation of the human BIN1 gene has been reported in ~40% Note of cases of metastatic prostate cancer. Bin1 is Mouse genetic studies indicate that Bin1 loss expressed in most primary tumors, even at slightly increases the general incidence of chronic elevated levels relative to benign tissues, but it is inflammation in the heart, pancreas, liver, and frequently grossly attenuated in expression or prostate. inactivated by aberrant splicing in metastatic tumors and androgen-independent tumor cell lines. Female fecundity Ectopic expression suppresses the growth of Note prostate cancer cell lines in vitro. Mouse genetic Mouse genetic studies indicate that mosaic loss of studies indicate that Bin1 loss is associated with an Bin1 increases reproductive physiology during increased incidence of prostate inflammation, aging. atrophy, hypertrophy, and intraepithelial neoplasia Specifically, female mosaic mice exhibit extended during aging (Ge et al., 2000b). fecundity during aging, retaining reproductive Neuroblastoma capability ~6 months longer than control mice Oncogenesis (Chang et al., 2007a). Neuroblastoma (NB) is the most common solid Alzheimer's disease tumor of childhood and is responsible for 15% of Note childhood cancer-related deaths. Bin1 expression is grossly reduced in MYCN amplified and metastatic BIN1 transcript levels are increased in AD brains NB compared with MYCN single-copy and (Tan et al., 2013). Genome-wide association studies localized NB as evaluated by real-time RT-PCR. (GWAS) have identified the BIN1 gene as the most Enforced expression of Bin1 in MYCN amplified important genetic susceptibility locus in human NB cell lines markedly inhibits colony Alzheimer's disease (AD) after APOE. Decreased formation (Hogarty et al., 2000). expression of the Drosophila BIN1 suppress Tau- mediated neurotoxicity. Tau and BIN1 has been Cardiomyopathy demonstrated to colocalize and interact in human Note neuroblastoma cells and in mouse brain (Chapuis et Dilated cardiomyopathy (DCM) is a leading cause al., 2013; Kingwell, 2013). of heart failure with as much as >25% of cases of familial etiology. A whole genome screen References performed in a three-generation family with 12 Sakamuro D, Elliott KJ, Wechsler-Reya R, Prendergast affected individuals with autosomal dominant GC. BIN1 is a novel MYC-interacting protein with features familial DCM defined linkage to chromosome of a tumour suppressor. Nat Genet. 1996 Sep;14(1):69-77 2q14-q22 where the human BIN1 gene is located Butler MH, David C, Ochoa GC, Freyberg Z, Daniell L, (Jung et al., 1999). While BIN1 was not specifically Grabs D, Cremona O, De Camilli P. Amphiphysin II identified as the germane locus, mouse studies (SH3P9; BIN1), a member of the amphiphysin/Rvs family, indicate that genetic ablation causes is concentrated in the cortical cytomatrix of axon initial segments and nodes of ranvier in brain and around T cardiomyopathy during development. Moreover, in tubules in skeletal muscle. J Cell Biol. 1997 Jun mice where Bin1 is ablated after birth in a tissue- 16;137(6):1355-67 specific manner in cardiomyocytes, a progressive Wechsler-Reya R, Sakamuro D, Zhang J, Duhadaway J, cardiomyopathy develops consistent with the Prendergast GC. Structural analysis of the human BIN1 possibility that Bin1 loss of function may be a gene. Evidence for tissue-specific transcriptional regulation cause of DCM (Muller et al., 2003). and alternate RNA splicing. J Biol Chem. 1997 Dec 12;272(50):31453-8 Centronuclear myopathy Wechsler-Reya RJ, Elliott KJ, Prendergast GC. A role for Note the putative tumor suppressor Bin1 in muscle cell Germ-line mutations of Bin1 are associated with differentiation. Mol Cell Biol. 1998 Jan;18(1):566-75 formation of this rare familial disorder Balguerie A, Sivadon P, Bonneu M, Aigle M. Rvs167p, the characterized by abnormal centralization of nuclei budding yeast homolog of amphiphysin, colocalizes with actin patches. J Cell Sci. 1999 Aug;112 ( Pt 15):2529-37 in muscle fibers and severe muscle weakness (Nicot et al., 2007; Smith et al., 2014). In five affected Elliott K, Sakamuro D, Basu A, Du W, Wunner W, Staller individuals studied from three non-sanguineous P, Gaubatz S, Zhang H, Prochownik E, Eilers M, Prendergast GC. Bin1 functionally interacts with Myc and families, two mutations extinguishing the inhibits cell proliferation via multiple mechanisms. membrane binding activity of the BAR domain Oncogene. 1999 Jun 17;18(24):3564-73

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Ge K, DuHadaway J, Du W, Herlyn M, Rodeck U, bin1/amphiphysin2 accentuates the neoplastic character of Prendergast GC. Mechanism for elimination of a tumor transformed mouse fibroblasts. Cancer Biol Ther. 2004 suppressor: aberrant splicing of a brain-specific exon Dec;3(12):1236-42 causes loss of function of Bin1 in melanoma. Proc Natl Acad Sci U S A. 1999 Aug 17;96(17):9689-94 Muller AJ, DuHadaway JB, Donover PS, Sutanto-Ward E, Prendergast GC. Inhibition of indoleamine 2,3- Jung M, Poepping I, Perrot A, Ellmer AE, Wienker TF, dioxygenase, an immunoregulatory target of the cancer Dietz R, Reis A, Osterziel KJ. Investigation of a family with suppression gene Bin1, potentiates cancer chemotherapy. autosomal dominant dilated cardiomyopathy defines a Nat Med. 2005 Mar;11(3):312-9 novel locus on chromosome 2q14-q22. Am J Hum Genet. 1999 Oct;65(4):1068-77 Ren G, Vajjhala P, Lee JS, Winsor B, Munn AL. The BAR domain proteins: molding membranes in fission, fusion, Elliott K, Ge K, Du W, Prendergast GC. The c-Myc- and phagy. Microbiol Mol Biol Rev. 2006 Mar;70(1):37-120 interacting adaptor protein Bin1 activates a caspase- independent cell death program. Oncogene. 2000 Sep Chang MY, Boulden J, Katz JB, Wang L, Meyer TJ, Soler 28;19(41):4669-84 AP, Muller AJ, Prendergast GC. Bin1 ablation increases susceptibility to cancer during aging, particularly lung Ge K, Duhadaway J, Sakamuro D, Wechsler-Reya R, cancer. Cancer Res. 2007a Aug 15;67(16):7605-12 Reynolds C, Prendergast GC. Losses of the tumor suppressor BIN1 in breast carcinoma are frequent and Chang MY, Boulden J, Sutanto-Ward E, Duhadaway JB, reflect deficits in programmed cell death capacity. Int J Soler AP, Muller AJ, Prendergast GC. Bin1 ablation in Cancer. 2000a Feb 1;85(3):376-83 mammary gland delays tissue remodeling and drives cancer progression. Cancer Res. 2007b Jan 1;67(1):100-7 Ge K, Minhas F, Duhadaway J, Mao NC, Wilson D, Buccafusca R, Sakamuro D, Nelson P, Malkowicz SB, Kuznetsova EB, Kekeeva TV, Larin SS, Zemlyakova VV, Tomaszewski J, Prendergast GC. Loss of heterozygosity Khomyakova AV, Babenko OV, Nemtsova MV, Zaletayev and tumor suppressor activity of Bin1 in prostate DV, Strelnikov VV. Methylation of the BIN1 gene promoter carcinoma. Int J Cancer. 2000b Apr 15;86(2):155-61 CpG island associated with breast and prostate cancer. J Carcinog. 2007 May 4;6:9 Hogarty MD, Liu X, Thompson PM, White PS, Sulman EP, Maris JM, Brodeur GM. BIN1 inhibits colony formation and Nicot AS, Toussaint A, Tosch V, Kretz C, Wallgren- induces apoptosis in neuroblastoma cell lines with MYCN Pettersson C, Iwarsson E, Kingston H, Garnier JM, amplification. Med Pediatr Oncol. 2000 Dec;35(6):559-62 Biancalana V, Oldfors A, Mandel JL, Laporte J. Mutations in amphiphysin 2 (BIN1) disrupt interaction with dynamin 2 Lee E, Marcucci M, Daniell L, Pypaert M, Weisz OA, and cause autosomal recessive centronuclear myopathy. Ochoa GC, Farsad K, Wenk MR, De Camilli P. Nat Genet. 2007 Sep;39(9):1134-9 Amphiphysin 2 (Bin1) and T-tubule biogenesis in muscle. Science. 2002 Aug 16;297(5584):1193-6 Ramalingam A, Prendergast GC. Bin1 homolog hob1 supports a Rad6-Set1 pathway of transcriptional DuHadaway JB, Du W, Donover S, Baker J, Liu AX, Sharp repression in fission yeast. Cell Cycle. 2007 Jul DM, Muller AJ, Prendergast GC. Transformation-selective 1;6(13):1655-62 apoptotic program triggered by farnesyltransferase inhibitors requires Bin1. Oncogene. 2003 Jun Prendergast GC, Muller AJ, Ramalingam A, Chang MY. 5;22(23):3578-88 BAR the door: cancer suppression by amphiphysin-like genes. Biochim Biophys Acta. 2009 Jan;1795(1):25-36 DuHadaway JB, Lynch FJ, Brisbay S, Bueso-Ramos C, Troncoso P, McDonnell T, Prendergast GC. Hong TT, Smyth JW, Gao D, Chu KY, Vogan JM, Fong Immunohistochemical analysis of Bin1/Amphiphysin II in TS, Jensen BC, Colecraft HM, Shaw RM. BIN1 localizes human tissues: diverse sites of nuclear expression and the L-type calcium channel to cardiac T-tubules. PLoS losses in prostate cancer. J Cell Biochem. 2003 Feb Biol. 2010 Feb 16;8(2):e1000312 15;88(3):635-42 Lundgaard GL, Daniels NE, Pyndiah S, Cassimere EK, Humbert P, Russell S, Richardson H. Dlg, Scribble and Lgl Ahmed KM, Rodrigue A, Kihara D, Post CB, Sakamuro D. in cell polarity, cell proliferation and cancer. Bioessays. Identification of a novel effector domain of BIN1 for cancer 2003 Jun;25(6):542-53 suppression. J Cell Biochem. 2011 Oct;112(10):2992-3001 Leprince C, Le Scolan E, Meunier B, Fraisier V, Brandon Chang MY, Boulden J, Valenzano MC, Soler AP, Muller N, De Gunzburg J, Camonis J. Sorting nexin 4 and AJ, Mullin JM, Prendergast GC. Bin1 attenuation amphiphysin 2, a new partnership between endocytosis suppresses experimental colitis by enforcing intestinal and intracellular trafficking. J Cell Sci. 2003 May 15;116(Pt barrier function. Dig Dis Sci. 2012 Jul;57(7):1813-21 10):1937-48 Hong TT, Cogswell R, James CA, Kang G, Pullinger CR, Muller AJ, Baker JF, DuHadaway JB, Ge K, Farmer G, Malloy MJ, Kane JP, Wojciak J, Calkins H, Scheinman Donover PS, Meade R, Reid C, Grzanna R, Roach AH, MM, Tseng ZH, Ganz P, De Marco T, Judge DP, Shaw Shah N, Soler AP, Prendergast GC. Targeted disruption of RM. Plasma BIN1 correlates with heart failure and predicts the murine Bin1/Amphiphysin II gene does not disable arrhythmia in patients with arrhythmogenic right ventricular endocytosis but results in embryonic cardiomyopathy with cardiomyopathy. Heart Rhythm. 2012 Jun;9(6):961-7 aberrant myofibril formation. Mol Cell Biol. 2003 Tanida S, Mizoshita T, Ozeki K, Tsukamoto H, Kamiya T, Jun;23(12):4295-306 Kataoka H, Sakamuro D, Joh T. Mechanisms of Cisplatin- Routhier EL, Donover PS, Prendergast GC. hob1+, the Induced Apoptosis and of Cisplatin Sensitivity: Potential of fission yeast homolog of Bin1, is dispensable for BIN1 to Act as a Potent Predictor of Cisplatin Sensitivity in endocytosis or actin organization, but required for the Gastric Cancer Treatment. Int J Surg Oncol. response to starvation or genotoxic stress. Oncogene. 2012;2012:862879 2003 Feb 6;22(5):637-48 Chapuis J, Hansmannel F, Gistelinck M, Mounier A, Van Muller AJ, DuHadaway JB, Donover PS, Sutanto-Ward E, Cauwenberghe C, Kolen KV, Geller F, Sottejeau Y, Harold Prendergast GC. Targeted deletion of the suppressor gene D, Dourlen P, Grenier-Boley B, Kamatani Y, Delepine B,

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 83 BIN1 (bridging integrator 1) Thomas S, et al.

Demiautte F, Zelenika D, Zommer N, Hamdane M, Tan MS, Yu JT, Tan L. Bridging integrator 1 (BIN1): form, Bellenguez C, Dartigues JF, Hauw JJ, Letronne F, Ayral function, and Alzheimer's disease. Trends Mol Med. 2013 AM, Sleegers K, Schellens A, Broeck LV, Engelborghs S, Oct;19(10):594-603 De Deyn PP, Vandenberghe R, O'Donovan M, Owen M, Epelbaum J, Mercken M, Karran E, Bantscheff M, Drewes Smith LL, Gupta VA, Beggs AH. Bridging integrator 1 G, Joberty G, Campion D, Octave JN, Berr C, Lathrop M, (Bin1) deficiency in zebrafish results in centronuclear Callaerts P, Mann D, Williams J, Buée L, Dewachter I, Van myopathy. Hum Mol Genet. 2014 Jul 1;23(13):3566-78 Broeckhoven C, Amouyel P, Moechars D, Dermaut B, Wu T, Shi Z, Baumgart T. Mutations in BIN1 associated Lambert JC. Increased expression of BIN1 mediates with centronuclear myopathy disrupt membrane Alzheimer genetic risk by modulating tau pathology. Mol remodeling by affecting protein density and Psychiatry. 2013 Nov;18(11):1225-34 oligomerization. PLoS One. 2014;9(4):e93060 Kingwell K. Alzheimer disease: BIN1 variant increases risk of Alzheimer disease through tau. Nat Rev Neurol. 2013 This article should be referenced as such: Apr;9(4):184 Thomas S, Chang MY, Prendergast GC. BIN1 (bridging integrator 1). Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2):78-84.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 84

Atlas of Genetics and Cytogenetics

in Oncology and Haematology

OPEN ACCESS JOURNAL INIST -CNRS

Gene Section Review

CDH3 (Cadherin 3, Type 1, P-Cadherin (Placental)) André Filipe Vieira, Ana Sofia Ribeiro, Joana Paredes Institute of Pathology and Immunology of the University of Porto, Portugal (AFV, ASR, JP)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/CDH3ID40025ch16q22.html DOI: 10.4267/2042/56291 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

(CCDS10868.1). Abstract Concerning CDH3/P-cadherin gene regulation, the Review on CDH3, with data on DNA/RNA, on the main transcriptional activators described for the protein encoded and where the gene is implicated. CDH3/P-cadherin gene promoter are β-catenin (Faraldo et al., 2007), p63 (Shimomura et al., 2008) Identity and C/EBP β (Albergaria et al., 2010; Albergaria et al., 2013). Other names: CDHP, HJMD, PCAD In contrast, BRCA1/c-Myc/Sp1 complex acts as a HGNC (Hugo): CDH3 transcriptional repressor of the CDH3 promoter Location: 16q22.1 (Gorski et al., 2009). It was also demonstrated that ER can indirectly DNA/RNA repress P-cadherin expression by promoting epigenetic changes in the CDH3 gene promoter Description (Paredes et al., 2004; Albergaria et al., 2010). DNA contains 54807 bp containing 16 coding This gene has 12 transcripts (splice variants), of exons. which 5 are protein coding transcripts, 4 are transcripts suffering nonsense mediated decay, and Transcription 3 transcripts do not code for any protein product 4276 bp mRNA transcribed in centromeric to (ensemble ENSG00000062038 and vega genome telomeric orientation; 2490 bp open reading frame OTTHUMG00000137560).

Localization of CDH3 gene (P-cadherin). 5 cadherin genes (CDH1; CDH3; CDH5; CDH8; and CDH11) are clustered in the 16q21-q22.1 region. The CDH3 gene is localized in the larger arm of chromosome 16, just 32Kb upstream of the gene encoding CDH1 (E-cadherin) (Bussemakers et al., 1994; Kremmidiotis et al., 1998).

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The genomic structure of the CDH3/P-cadherin gene is constituted by 16 coding exons (NCBI Reference Sequence: NM_001793.4): the extracellular part of P-cadherin is encoded by 10 exons (exons 4-13), whereas the transmembrane and the intracellular domains are codified by the last 3 exons (exons 14-16) (Albergaria et al., 2011) (NCBI Reference Sequence: NM_001793.4).

Protein Description Described for the first time in 1986, as "a novel class of cadherin that appeared in developing mouse embryos", this adhesion molecule was found in the tissues that gave rise to its name, the placenta (Nose and Takeichi, 1986). P-cadherin is a transmembrane glycoprotein that belongs to a large family of molecules that mediate calcium-dependent homophilic cell-cell adhesion. It plays a role in many cellular processes such as embryonic development, differentiation, cell polarity, growth and migration (Larue et al., 1996). P-cadherin is composed by three domains: 1) an extracellular portion responsible for calcium- dependent homotypic cadherin-cadherin interaction (which has 5 repeated cadherin domains); 2) a single pass transmembrane domain; and 3) a highly conserved cytoplasmic domain that binds to the intracellular catenins p120-catenin and β-catenin. Catenins have a dual role, acting as signalling mediators or as adaptor molecules that stabilize the cadherin complex at the membrane and link the cadherin molecule to the actin filaments of the cytoskeleton (Wheelock et al., 2001). Expression It is expressed in the placenta of mice (hence, its name). It is also expressed in human placental tissues, albeit at lower levels (Shimoyama et al., 1989; Sahin et al., 2014). Despite being expressed in several human fetal structures, in the adult it is only expressed in certain tissues, usually co-expressed with E-cadherin, such as the basal layer of the epidermis, the breast and the prostate, as well as the mesothelium, the ovary, Image of a classical type I cadherin, adapted with the hair follicle, and the corneal endothelium (Nose permission from the RCSB PDB March 2008 Molecule of and Takeichi, 1986; Imai et al., 2008). the Month feature by David Goodsell (doi: 10.2210/rcsb_pdb/mom_2008_3). P-cadherin is a According to human protein reference database transmembrane protein included in the classical cadherin (HPRD:00227), the major sites of expression family, with an ectodomain containing 5 cadherin repeats include endometrium, the glomerulus, hair follicle, (which interacts with another cadherin's ectodomain in a cis keratinocytes, mammary myoepithelium, or trans manner) and a highly conserved cytoplasmic domain that binds to catenins. Sharing about 67% of melanocytes, oocytes, spermatozoa, placenta, homology with the CDH1/E-cadherin gene, P-cadherin prostate, retina, serum, skin. An 80 KDa fragment differs mainly in the extracellular portion and it is far less of P-cadherin (known as soluble P-cadherin) is also characterized. found in human breast milk (Soler et al., 2002),

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 86 CDH3 (cadherin 3, type 1, P-cadherin (placental)) Vieira AF, et al.

nipple aspirates (Mannello et al., 2008), semen (De Homology Paul et al., 2005) and urine (Adachi et al., 2006). Sharing about 67% of homology with the CDH1/E- Localisation cadherin gene, P-cadherin differs mainly in the Cell junctions: a single-pass type I membrane extracellular portion and it is far less characterized. protein anchored to actin microfilaments through 64 organisms have orthologs with the human CDH3 association with α-catenin, β-catenin and γ-catenin. gene. For example, the CDH3 gene is conserved in Sequential proteolysis induced by apoptosis or chimpanzee, dog, cow, mouse, rat and chicken calcium influx, results in translocation from sites of (HomoloGene:20425). cell-cell contact to the cytoplasm. Function Mutations Cell-cell adhesion: cadherin mediated cell-cell Note adhesion is accomplished by homophilic According to the Human Gene Mutation Database, interactions between two cadherin molecules at the 21 mutations have been described for the P- surface of the respective cells in a cis and/or trans cadherin/CDH3 gene, namely 9 missense/nonsense manner (Cavallaro and Dejana, 2011) and the mutations, 4 splicing mutations, 1 regulatory cadherin-catenin complex constitutes the main mutation, 1 gross deletion, 5 small deletions and 1 building block of the adherens-type junctions. small insertion (The Human Gene Mutation These complexes also represent a major regulatory Database). There are no reported descriptions of mechanism that guides cell fate decisions, small indels, gross insertions/duplications, complex influencing cell growth, differentiation, cell rearrangements or repeat variations. 16 mutations motility and survival (Cavallaro and Dejana, 2011). are associated with Hypotricosis with Juvenile Cell signalling: P-cadherin shares common Macular Dystrophy (HJMD) and 2 mutations are interplayers with the wnt signalling pathway (eg. : implicated with Ectodermal dysplasia, Ectrodactyly β-catenin) (Kamposioras et al., 2013; Samuelov et and Macular dystrophy (EEM) syndrome (The al., 2013) and Integrin signalling (Vieira et al., Human Gene Mutation Database). 2014). In cancer, it may have a tumour suppressive Regarding polymorphisms, several SNPs have been or a cancer promoting function, depending on the reported for the CDH3 gene that have no clinical cellular and tissue context (see below). significance because they code for synonymous codons or related residues (ClinVar).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 87 CDH3 (cadherin 3, type 1, P-cadherin (placental)) Vieira AF, et al.

Summary of the human CDH3 gene mutations. 21 mutations have been described for the P-cadherin/CDH3 gene, namely 9 missense/nonsense mutations, 4 splicing mutations, 1 regulatory mutation, 5 small deletions, 1 small insertion and 1 gross deletion (The Human Gene Mutation Database).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 88 CDH3 (cadherin 3, type 1, P-cadherin (placental)) Vieira AF, et al.

Germinal increased stem cell activity (Vieira et al., 2012) and cross-talk with integrin oncogenic signalling Human germline mutations for the CDH3 gene pathways (Vieira et al., 2014). have been reported to carry the phenotype of HJMD P-cadherin up-regulation is predominantly found in and EEM syndromes (Sprecher et al., 2001;Kjaer et the basal-like subgroup of breast cancers (Matos et al., 2005; Avitan-Hersh et al., 2012; Halford et al., al., 2005; Paredes et al., 2007a; Paredes et al., 2012). 2007b) and it is strongly associated with the The germline deletion of CDH3 in the mouse presence of BRCA1 mutation (Arnes et al., 2005) causes breast secretory immaturity and premature and poor clinical outcome (Paredes et al., 2005; mammary gland differentiation. Turashvili et al., 2011). It has been proposed that P- The P-cadherin mutant mice develop hyperplasia cadherin in conjugation with vimentin and CK14 and dysplasia of the mammary epithelium with age constitutes a better criterion for the identification of and, in contrast to humans, no reports regarding basal-like breast carcinomas by development defects have been described (Radice immunohistochemistry (Sousa et al., 2010). et al., 1997). Prognosis Implicated in P-cadherin overexpression in breast cancer is an independent factor of poor prognosis (poor disease Various cancers free and overall survival) (Paredes et al., 2005; Note Turashvili et al., 2011). P-cadherin is altered in various human tumours, but Hypotrichosis with juvenile macular its effective role in the carcinogenesis process dystrophy (HJMD) remains object of debate, since it can behave differently depending on the studied tumour cell Note model and context. In humans, the loss of P-cadherin induces For example, in melanoma, P-cadherin seems to characteristic genetic syndromes. have a tumour suppressive function, exactly as E- CDH3 gene mutations have been shown to cause P- cadherin (Van Marck et al., 2005). cadherin functional inactivation, leading to In breast cancer P-cadherin is often overexpressed developmental defects associated with and it is reported to exhibit tumour promoting hypotrichosis with juvenile macular dystrophy effects (Paredes et al., 2012). (HJMD) (Sprecher et al., 2001; Avitan-Hersh et al., Importantly, P-cadherin upregulation is also found 2012; Halford et al., 2012). in other malignancies such as gastric, endometrial, Disease colorectal and pancreatic carcinomas (Hardy et al., Hypotrichosis with juvenile macular dystrophy 2002; Stefansson et al., 2004; Taniuchi et al., 2005; (HJMD; OMIM: 601553) is a rare recessive Imai et al., 2008). disorder, characterized by hair loss heralding Breast cancer progressive macular degeneration and early Note blindness. Affected HJMD individuals are born P-cadherin aberrant expression is found in 20% to with seemingly normal hair but develop alopecia of 40% of invasive breast carcinomas, as well as in the scalp at around 3 months. After the age of 3 25% of pre-invasive (in situ) ductal carcinomas. years, affected individuals develop progressive Aberrant P-cadherin expression was shown to be macular degeneration with slight peripheral retinal associated with tumours of high histological grade, dystrophy. The severe degenerative changes of the as well as with well established markers of poor retinal macula culminate in blindness during the prognosis, like Ki-67, EGFR, CK5, vimentin, p53 second to third decade of life. Since Sprech er et al. and HER-2 expression, and negatively associated (2001) first established a link between this disease with age at prognosis and hormonal receptors (ER and a mutation in gene encoding CDH3/P-cadherin and PgR) expression. (Sprecher et al., 2001), several other mutations None of these reports showed a significant were found, which essentially disturb the Ca2+ association with tumour size and lymph node binding and the cadherin functional domains or metastasis (Turashvili et al., 2011; Peralta Soler et result in the synthesis of a truncated form of P- al., 1999; Gamallo et al., 2001; Paredes et al., 2002; cadherin or in the absence of P-cadherin expression. Paredes et al., 2005). Cytogenetics P-cadherin aggressive behaviour in breast cancer is The following allelic variants are responsible for dependent on an E-cadherin positive background HJMD: (Ribeiro et al., 2013) and it is substantially - CDH3, c981delG - (Sprecher et al., 2001) attributed to an increased migratory and invasive - CDH3, Arg503His - (Indelman et al., 2002) capacity of cancer cells (Ribeiro et al., 2010), - CDH3, Leu168Term - (Indelman et al., 2003)

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 89 CDH3 (cadherin 3, type 1, P-cadherin (placental)) Vieira AF, et al.

- CDH3, Arg221Term - (Indelman et al., 2007) Shimoyama Y, Yoshida T, Terada M, Shimosato Y, Abe O, - CDH3, Tyr249Term - (Avitan-Hersh et al., 2012) Hirohashi S. Molecular cloning of a human Ca2+- dependent cell-cell adhesion molecule homologous to - CDH3, Glu504Lys - (Indelman et al., 2007) mouse placental cadherin: its low expression in human - CDH3, His575Arg - (Indelman et al., 2007) placental tissues. J Cell Biol. 1989 Oct;109(4 Pt 1):1787- - CDH3, Tyr615Term - (Indelman et al., 2005) 94 - CDH3, IVS2 ds G-A +1 - (Indelman et al., 2007) Bussemakers MJ, van Bokhoven A, Völler M, Smit FP, - CDH3, IVS10 as G-A -1 - (Jelani et al., 2009; Schalken JA. The genes for the calcium-dependent cell Kamran-ul-Hassan Naqvi et al., 2010) adhesion molecules P- and E-cadherin are tandemly arranged in the human genome. Biochem Biophys Res - CDH3, IVS12 as A-G -2 - (Shimomura et al., Commun. 1994 Sep 15;203(2):1291-4 2010) - CDH3, c.462delT - (Indelman et al., 2003) Larue L, Antos C, Butz S, Huber O, Delmas V, Dominis M, Kemler R. A role for cadherins in tissue formation. - CDH3, c.2117delG - (Indelman et al., 2003) Development. 1996 Oct;122(10):3185-94 - CDH3, gDNA 8815bp deleted incl exons 12-13 - (Halford et al., 2012) Radice GL, Ferreira-Cornwell MC, Robinson SD, Rayburn H, Chodosh LA, Takeichi M, Hynes RO. Precocious Ectodermal dysplasia, ectrodactyly mammary gland development in P-cadherin-deficient mice. J Cell Biol. 1997 Nov 17;139(4):1025-32 and macular dystrophy (EEM) Kremmidiotis G, Baker E, Crawford J, Eyre HJ, Nahmias J, Note Callen DF. Localization of human cadherin genes to CDH3 gene mutations have been shown to cause P- chromosome regions exhibiting cancer-related loss of cadherin functional inactivation, leading to heterozygosity. Genomics. 1998 May 1;49(3):467-71 ectodermal dysplasia, ectrodactyly, and macular Peralta Soler A, Knudsen KA, Salazar H, Han AC, dystrophy (EEM syndrome ), a developmental Keshgegian AA. P-cadherin expression in breast defect associated syndrome. This inherited disease carcinoma indicates poor survival. Cancer. 1999 Oct 1;86(7):1263-72 is characterized by sparse hair and macular dystrophy of the retina, and split hand/foot Gamallo C, Moreno-Bueno G, Sarrió D, Calero F, Hardisson D, Palacios J. The prognostic significance of P- malformation (Kjaer et al., 2005). cadherin in infiltrating ductal breast carcinoma. Mod Disease Pathol. 2001 Jul;14(7):650-4 This ectodermal defect is characterised by Sprecher E, Bergman R, Richard G, Lurie R, Shalev S, hypotrichosis with sparse and short hair on the Petronius D, Shalata A, Anbinder Y, Leibu R, Perlman I, scalp, sparse and short eyebrows and eyelashes, and Cohen N, Szargel R. Hypotrichosis with juvenile macular dystrophy is caused by a mutation in CDH3, encoding P- partial anodontia. Different degrees of absence cadherin. Nat Genet. 2001 Oct;29(2):134-6 deformities as well as syndactyly have been described, the hands often being more severely Wheelock MJ, Soler AP, Knudsen KA. Cadherin junctions in mammary tumors. J Mammary Gland Biol Neoplasia. affected than the feet. The retinal lesion appears as 2001 Jul;6(3):275-85 a central geographic atrophy of the retinal pigment epithelium and choriocapillary layer of the macular Hardy RG, Tselepis C, Hoyland J, Wallis Y, Pretlow TP, Talbot I, Sanders DS, Matthews G, Morton D, Jankowski area with coarse hyperpigmentations and sparing of JA. Aberrant P-cadherin expression is an early event in the larger choroidal vessels. hyperplastic and dysplastic transformation in the colon. Kjaer et al. (2005) first established a link between Gut. 2002 Apr;50(4):513-9 families with ectodermal dysplasia, ectrodactyly, Indelman M, Bergman R, Lurie R, Richard G, Miller B, and macular dystrophy (EEM; OMIM: 225280) and Petronius D, Ciubutaro D, Leibu R, Sprecher E. A homozygous mutations in CDH3/P-cadherin in missense mutation in CDH3, encoding P-cadherin, causes hypotrichosis with juvenile macular dystrophy. J Invest affected individuals: a missense mutation Dermatol. 2002 Nov;119(5):1210-3 (114021.0003) and a deletion (114021.0004), respectively (Kjaer et al., 2005). Paredes J, Milanezi F, Viegas L, Amendoeira I, Schmitt F. P-cadherin expression is associated with high-grade ductal Cytogenetics carcinoma in situ of the breast. Virchows Arch. 2002 The following allelic variants are responsible for Jan;440(1):16-21 EEM: Soler AP, Russo J, Russo IH, Knudsen KA. Soluble - CDH3, Asn322Ile - (Kjaer et al., 2005) fragment of P-cadherin adhesion protein found in human - CDH3, c.829delG - (Kjaer et al., 2005) milk. J Cell Biochem. 2002;85(1):180-4 - CDH3, Gly277Val - (Basel-Vanagaite et al., Indelman M, Hamel CP, Bergman R, Nischal KK, 2010) Thompson D, Surget MO, Ramon M, Ganthos H, Miller B, Richard G, Lurie R, Leibu R, Russell-Eggitt I, Sprecher E. Phenotypic diversity and mutation spectrum in References hypotrichosis with juvenile macular dystrophy. J Invest Dermatol. 2003 Nov;121(5):1217-20 Nose A, Takeichi M. A novel cadherin cell adhesion molecule: its expression patterns associated with Paredes J, Stove C, Stove V, Milanezi F, Van Marck V, implantation and organogenesis of mouse embryos. J Cell Derycke L, Mareel M, Bracke M, Schmitt F. P-cadherin is Biol. 1986 Dec;103(6 Pt 2):2649-58 up-regulated by the antiestrogen ICI 182,780 and

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 90 CDH3 (cadherin 3, type 1, P-cadherin (placental)) Vieira AF, et al.

promotes invasion of human breast cancer cells. Cancer Nakamura Y, Baba H, Nishimura Y. Identification of a Res. 2004 Nov 15;64(22):8309-17 novel tumor-associated antigen, cadherin 3/P-cadherin, as a possible target for immunotherapy of pancreatic, gastric, Stefansson IM, Salvesen HB, Akslen LA. Prognostic and colorectal cancers. Clin Cancer Res. 2008 Oct impact of alterations in P-cadherin expression and related 15;14(20):6487-95 cell adhesion markers in endometrial cancer. J Clin Oncol. 2004 Apr 1;22(7):1242-52 Mannello F, Tonti GA, Medda V, Pederzoli A, Sauter ER. Increased shedding of soluble fragments of P-cadherin in Arnes JB, Brunet JS, Stefansson I, Bégin LR, Wong N, nipple aspirate fluids from women with breast cancer. Chappuis PO, Akslen LA, Foulkes WD. Placental cadherin Cancer Sci. 2008 Nov;99(11):2160-9 and the basal epithelial phenotype of BRCA1-related breast cancer. Clin Cancer Res. 2005 Jun 1;11(11):4003- Shimomura Y, Wajid M, Shapiro L, Christiano AM. P- 11 cadherin is a p63 target gene with a crucial role in the developing human limb bud and hair follicle. Development. De Paul AL, Bonaterra M, Soler AP, Knudsen KA, Roth 2008 Feb;135(4):743-53 FD, Aoki A. Soluble p-cadherin found in human semen. J Androl. 2005 Jan-Feb;26(1):44-7 Jelani M, Salman Chishti M, Ahmad W. A novel splice-site mutation in the CDH3 gene in hypotrichosis with juvenile Indelman M, Leibu R, Jammal A, Bergman R, Sprecher E. macular dystrophy. Clin Exp Dermatol. 2009 Jan;34(1):68- Molecular basis of hypotrichosis with juvenile macular 73 dystrophy in two siblings. Br J Dermatol. 2005 Sep;153(3):635-8 Albergaria A, Ribeiro AS, Pinho S, Milanezi F, Carneiro V, Sousa B, Sousa S, Oliveira C, Machado JC, Seruca R, Kjaer KW, Hansen L, Schwabe GC, Marques-de-Faria AP, Paredes J, Schmitt F. ICI 182,780 induces P-cadherin Eiberg H, Mundlos S, Tommerup N, Rosenberg T. Distinct overexpression in breast cancer cells through chromatin CDH3 mutations cause ectodermal dysplasia, ectrodactyly, remodelling at the promoter level: a role for C/EBPbeta in macular dystrophy (EEM syndrome). J Med Genet. 2005 CDH3 gene activation. Hum Mol Genet. 2010 Jul Apr;42(4):292-8 1;19(13):2554-66 Matos I, Dufloth R, Alvarenga M, Zeferino LC, Schmitt F. Basel-Vanagaite L, Pasmanik-Chor M, Lurie R, Yeheskel p63, cytokeratin 5, and P-cadherin: three molecular A, Kjaer KW. CDH3-Related Syndromes: Report on a New markers to distinguish basal phenotype in breast Mutation and Overview of the Genotype-Phenotype carcinomas. Virchows Arch. 2005 Oct;447(4):688-94 Correlations. Mol Syndromol. 2010;1(5):223-230 Paredes J, Albergaria A, Oliveira JT, Jerónimo C, Milanezi Gorski JJ, James CR, Quinn JE, Stewart GE, Staunton F, Schmitt FC. P-cadherin overexpression is an indicator of KC, Buckley NE, McDyer FA, Kennedy RD, Wilson RH, clinical outcome in invasive breast carcinomas and is Mullan PB, Harkin DP. BRCA1 transcriptionally regulates associated with CDH3 promoter hypomethylation. Clin genes associated with the basal-like phenotype in breast Cancer Res. 2005 Aug 15;11(16):5869-77 cancer. Breast Cancer Res Treat. 2010 Aug;122(3):721-31 Taniuchi K, Nakagawa H, Hosokawa M, Nakamura T, Kamran-ul-Hassan Naqvi S, Azeem Z, Ali G, Ahmad W. A Eguchi H, Ohigashi H, Ishikawa O, Katagiri T, Nakamura novel splice-acceptor site mutation in CDH3 gene in a Y. Overexpressed P-cadherin/CDH3 promotes motility of consanguineous family exhibiting hypotrichosis with pancreatic cancer cells by interacting with p120ctn and juvenile macular dystrophy. Arch Dermatol Res. 2010 activating rho-family GTPases. Cancer Res. 2005 Apr Nov;302(9):701-3 15;65(8):3092-9 Ribeiro AS, Albergaria A, Sousa B, Correia AL, Bracke M, Van Marck V, Stove C, Van Den Bossche K, Stove V, Seruca R, Schmitt FC, Paredes J. Extracellular cleavage Paredes J, Vander Haeghen Y, Bracke M. P-cadherin and shedding of P-cadherin: a mechanism underlying the promotes cell-cell adhesion and counteracts invasion in invasive behaviour of breast cancer cells. Oncogene. 2010 human melanoma. Cancer Res. 2005 Oct 1;65(19):8774- Jan 21;29(3):392-402 83 Shimomura Y, Wajid M, Kurban M, Christiano AM. Splice Adachi J, Kumar C, Zhang Y, Olsen JV, Mann M. The site mutations in the P-cadherin gene underlie human urinary proteome contains more than 1500 hypotrichosis with juvenile macular dystrophy. proteins, including a large proportion of membrane Dermatology. 2010;220(3):208-12 proteins. Genome Biol. 2006;7(9):R80 Sousa B, Paredes J, Milanezi F, Lopes N, Martins D, Faraldo MM, Teulière J, Deugnier MA, Birchmeier W, Dufloth R, Vieira D, Albergaria A, Veronese L, Carneiro V, Huelsken J, Thiery JP, Cano A, Glukhova MA. beta- Carvalho S, Costa JL, Zeferino L, Schmitt F. P-cadherin, Catenin regulates P-cadherin expression in mammary vimentin and CK14 for identification of basal-like basal epithelial cells. FEBS Lett. 2007 Mar 6;581(5):831-6 phenotype in breast carcinomas: an immunohistochemical Indelman M, Eason J, Hummel M, Loza O, Suri M, Leys study. Histol Histopathol. 2010 Aug;25(8):963-74 MJ, Bayne M, Schwartz FL, Sprecher E. Novel CDH3 Albergaria A, Ribeiro AS, Vieira AF, Sousa B, Nobre AR, mutations in hypotrichosis with juvenile macular dystrophy. Seruca R, Schmitt F, Paredes J. P-cadherin role in normal Clin Exp Dermatol. 2007 Mar;32(2):191-6 breast development and cancer. Int J Dev Biol. 2011;55(7- Paredes J, Correia AL, Ribeiro AS, Albergaria A, Milanezi 9):811-22 F, Schmitt FC. P-cadherin expression in breast cancer: a Cavallaro U, Dejana E. Adhesion molecule signalling: not review. Breast Cancer Res. 2007a;9(5):214 always a sticky business. Nat Rev Mol Cell Biol. 2011 Paredes J, Lopes N, Milanezi F, Schmitt FC. P-cadherin Mar;12(3):189-97 and cytokeratin 5: useful adjunct markers to distinguish Turashvili G, McKinney SE, Goktepe O, Leung SC, basal-like ductal carcinomas in situ. Virchows Arch. 2007b Huntsman DG, Gelmon KA, Los G, Rejto PA, Aparicio SA. Jan;450(1):73-80 P-cadherin expression as a prognostic biomarker in a 3992 Imai K, Hirata S, Irie A, Senju S, Ikuta Y, Yokomine K, case tissue microarray series of breast cancer. Mod Harao M, Inoue M, Tsunoda T, Nakatsuru S, Nakagawa H, Pathol. 2011 Jan;24(1):64-81

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Avitan-Hersh E, Indelman M, Khamaysi Z, Leibu R, Konstantaras C, Fountzilas G. The prognostic significance Bergman R. A novel nonsense CDH3 mutation in of WNT pathway in surgically-treated colorectal cancer: β- hypotrichosis with juvenile macular dystrophy. Int J catenin expression predicts for disease-free survival. Dermatol. 2012 Mar;51(3):325-7 Anticancer Res. 2013 Oct;33(10):4573-84 Halford S, Holt R, Németh AH, Downes SM. Homozygous Ribeiro AS, Sousa B, Carreto L, Mendes N, Nobre AR, deletion in CDH3 and hypotrichosis with juvenile macular Ricardo S, Albergaria A, Cameselle-Teijeiro JF, Gerhard dystrophy. Arch Ophthalmol. 2012 Nov;130(11):1490-2 R, Söderberg O, Seruca R, Santos MA, Schmitt F, Paredes J. P-cadherin functional role is dependent on E- Paredes J, Figueiredo J, Albergaria A, Oliveira P, Carvalho cadherin cellular context: a proof of concept using the J, Ribeiro AS, Caldeira J, Costa AM, Simões-Correia J, breast cancer model. J Pathol. 2013 Apr;229(5):705-18 Oliveira MJ, Pinheiro H, Pinho SS, Mateus R, Reis CA, Leite M, Fernandes MS, Schmitt F, Carneiro F, Figueiredo Samuelov L, Sprecher E, Sugawara K, Singh SK, Tobin C, Oliveira C, Seruca R. Epithelial E- and P-cadherins: role DJ, Tsuruta D, Bíró T, Kloepper JE, Paus R. Topobiology and clinical significance in cancer. Biochim Biophys Acta. of human pigmentation: P-cadherin selectively stimulates 2012 Dec;1826(2):297-311 hair follicle melanogenesis. J Invest Dermatol. 2013 Jun;133(6):1591-600 Vieira AF, Ricardo S, Ablett MP, Dionísio MR, Mendes N, Albergaria A, Farnie G, Gerhard R, Cameselle-Teijeiro JF, Sahin H, Akpak YK, Berber U, Gün I, Demirel D, Ergür AR. Seruca R, Schmitt F, Clarke RB, Paredes J. P-cadherin is Expression of P-cadherin (cadherin-3) and E-selectin in coexpressed with CD44 and CD49f and mediates stem cell the villous trophoblast of first trimester human placenta. J properties in basal-like breast cancer. Stem Cells. 2012 Turk Ger Gynecol Assoc. 2014;15(1):13-7 May;30(5):854-64 Vieira AF, Ribeiro AS, Dionísio MR, Sousa B, Nobre AR, Albergaria A, Resende C, Nobre AR, Ribeiro AS, Sousa B, Albergaria A, Santiago-Gómez A, Mendes N, Gerhard R, Machado JC, Seruca R, Paredes J, Schmitt F. Schmitt F, Clarke RB, Paredes J. P-cadherin signals CCAAT/enhancer binding protein β (C/EBP β) isoforms as through the laminin receptor α6β4 integrin to induce stem transcriptional regulators of the pro-invasive CDH3/P- cell and invasive properties in basal-like breast cancer cadherin gene in human breast cancer cells. PLoS One. cells. Oncotarget. 2014 Feb 15;5(3):679-92 2013;8(2):e55749 This article should be referenced as such: Kamposioras K, Konstantara A, Kotoula V, Lakis S, Vieira AF, Ribeiro AS, Paredes J. CDH3 (cadherin 3, type Kouvatseas G, Akriviadis E, Vrettou E, Dionysopoulos D, 1, P-cadherin (placental)). Atlas Genet Cytogenet Oncol Krikelis D, Papadopoulou K, Charalambous E, Chrisafi S, Haematol. 2015; 19(2):85-92.

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CELF2 (CUGBP, Elav-like family member 2) Satish Ramalingam, Shrikant Anant Department of Molecular and Integrative Physiology, Kansas University Medical Center, Kansas City, KS, USA (SR, SA)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/CELF2ID52815ch10p14.html DOI: 10.4267/2042/56292 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Location: 10p14 Note: Size: 331416 bases. Orientation: plus strand. CELF2 belongs to the family of RNA binding This gene is encoded by a gene located on proteins implicated in mRNA splicing, editing, chromosome 10p13-p14 between Généthon stability and translation. markers D10S547 and D10S223 (Li et al., 2001). This gene is encoded in a single large gene spanning over 159 kilo bases located on DNA/RNA chromosome 10 p13-p14 (between D10S547 and D10S223). Description This gene has 14 transcripts (splice variants) and The human CELF2 gene contains 14 exons the 3 major splice variants have distinct exon 1. spanning over approximately 159 kb of the This is an evolutionarily conserved ubiquitously genomic DNA. expressed protein. The members of the CELF protein family contain two N-terminal RNA Transcription recognition motif (RRM) domains and one C- Alternative promoters usage of CELF2 gene results terminal RRM domain, and a divergent segment of in three transcript variants, where the variants 2 and 160-230 amino acids between second and third 3 proteins have distinct exon 1 resulting in different RRM domains. This divergent domain is unique to 5' untranslated region (UTR) and have extended N- CELF2 proteins and has been shown to contain one terminal sequences (Ramalingam et al., 2008). or more activation molecules required for splicing There are totally 14 transcripts (splice variants) activity. CELF2 has been shown to bind to the reported so far. CUG and Au-rich element (ARE) in the target mRNA and shown to be implicated in muscular Protein dystrophy and cancer. Keywords Description RNA binding protein, mRNA stability, splicing, This is an evolutionarily conserved protein. The apoptosis, translation inhibition, muscular members of the CELF protein family contain two dystrophy, cancer N-terminal RNA recognition motif (RRM) domains and one C-terminal RRM domain, and a divergent Identity segment of 160-230 amino acids between second and third RRM domains. This divergent domain is Other names: BRUNOL3, CUGBP2, ETR-3, unique to CELF2 proteins and has been shown to ETR3, NAPOR contain one or more activation molecules required HGNC (Hugo): CELF2 for splicing activity (figure 1).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 93 CELF2 (CUGBP, Elav-like family member 2) Ramalingam S, Anant S

Figure 1. RRM position of CELF2 protein variants.

Expression untranslated region (3' UTR) of the target mRNAs. Upon binding to the AU-rich sequences in CELF2 is a ubiquitously expressed protein. cyclooxygenase-2 (COX-2) 3' UTR, CELF2 According to the NCBI Entrez GEO profiles the enhances the stability of COX-2 mRNA. However, CELF2 is expressed in brain, heart, thymus, spleen, CUGBP2 binding also results in the inhibition of its bone, tongue, stomach, intestine, pancreas, liver, translation (Murmu et al., 2004). In our earlier breast, lung, kidney, testis, ovary, prostate, placenta studies we have demonstrated that CELF2 can and skin. In addition, according to expression atlas interact with HuR, a key inducer of RNA stability brain, bone marrow, heart, spleen, lymph node, and translation, and competitively inhibit HuR ovary and adipose tissue has more expression of function (Sureban et al., 2007). Recently, platelet CELF2. derived growth factor was shown to enhance Localisation CELF2 binding to COX-2 mRNA through CELF2 variant 1 is predominantly nuclear, while increased phosphorylation of a tyrosine residue at variants 2 and 3 are predominantly cytoplasmic position 39 in the protein (Xu et al., 2007). These (Ramalingam et al., 2008). CELF2 variant 1 data suggest that posttranscriptional control accumulates in the cytoplasm following radiation mechanisms are in place to modulate the CELF2 exposure (Mukhopadhyay et al., 2003a). The C function as a regulator of stability and translation of terminus of CELF2 transcript variant 1 is rich in AU-rich transcripts. arginine and lysine residues 13 amino acids Homology (KRLKVQLKRSKND) 467 - 480, which is common for NLS elements recognized by importin According to GeneCards, the CELF2 has orthologs proteins. Ladd and Cooper, has reported that the C- in 72 species including much lower organisms such terminus of CELF2 contains a strong nuclear as Danio rerio, Drosophila melanogaster, localization signal overlapping the third RRM Caenorhabditis elegans, Xenopus tropicalis and (Ladd and Cooper, 2004). However, our Oryza sativa. Furthermore, in humans it has 6 unpublished data suggests that nuclear localization paralogs from CELF1 to CELF6. signal extends to the RNA recognition motif 1 and 2 domains. Finally, CELF2 has several leucine-rich Mutations motifs that resembles nuclear export signals Note recognized by the export protein CRM1. According to GeneCards, there is 7518 single Function nucleotide polymorphism. However, Ensembl CELF2 is an RNA-binding protein implicated in the reports that CELF2 has 7768 SNPs. In addition, the regulation of several post-transcriptional events. It Database of Genomic Variants shows that CELF2 has been shown to regulate pre-mRNA splicing has 18 structural variations. (Faustino and Cooper, 2005), mRNA editing (Anant et al., 2001), mRNA translation and Implicated in stability. CELF2 has been shown to be involved in alternative splicing of muscle specific genes Colon cancer including exon 5 of cardiac troponin T (Ladd et al., Note 2001), exon 11 of insulin receptor, intron 2 of Putative tumor suppressor CELF2 expression is chloride channel 1, exons 5 and 21 of NMDAR-1, consistently reduced during neoplastic and the muscle-specific exon of α-actinin (Gromak transformation suggesting that it might play a et al., 2003). Another function for CELF2 relates to crucial role in tumor initiation and progression of its ability to bind to AU-rich sequences in 3' colon cancer. In addition, CELF2 has been shown

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 94 CELF2 (CUGBP, Elav-like family member 2) Ramalingam S, Anant S

to induce mitotic catastrophic cell death in colon muscular atrophy (Anderson et al., 2004). Spinal cancer (Ramalingam et al., 2012). and bulbar muscular atrophy (SBMA) is an Pancreatic cancer inherited neurodegenerative disorder caused by the expansion of the polyglutamine (polyQ) tract of the Note androgen receptor (AR-polyQ). It has been shown Curcumin inhibits the pancreatic cancer growth by that miR-196a enhanced the decay of the AR inducing the expression of CELF2 thereby mRNA by silencing CUGBP, Elav-like family regulating the levels of cyclooxygenase 2 and member 2 (CELF2). CELF2 shown to directly act vascular endothelial growth factor expression on AR mRNA and enhance the stability of AR (Subramaniam et al., 2011). mRNA (Miyazaki et al., 2012). Myotonic Breast cancer dystrophy (DM) is a neuromuscular disorder associated with CTG triplet repeat expansion in the Note myotonin protein kinase gene (DMPK). It has been Breast cancer cells underwent apoptotic cell death suggested that the expanded CUG repeats sequester in response to radiation injury and this was reversed specific RNA-binding proteins and that such a by knockdown of CELF2 using specific siRNA sequestration results in abnormal RNA processing (Mukhopadhyay et al., 2003b). of several RNAs containing CUG repeats in Neuroblastoma multiple tissues affected in patients with DM. One Note of the members of the CUG-binding proteins, Colchicine treatment of neuroblastoma cells CUG-BP, has been identified previously (Lu et al., resulted in apoptotic cell death and CELF2 has been 1999). shown to be involved in the process of cell death Development (Li et al., 2001). Note Alzheimer's disease Overexpression of CELF2 by RNA microinjection Note resulted in severe defects in nervous system and It has been shown that variants in CUGBP2 on gastrulation, suggesting the need for tight control of chromosome 10p, are associated with AD in those napor gene regulation during embryo development highest-risk APOE e4 homozygotes. This (Choi et al., 2003). CELF2 appears to be an interaction observation is replicated in independent important factor for thymus development and is samples. CELF2 has one isoform that is expressed therefore a candidate gene for the thymus predominantly in neurons, and identification of hypoplasia/aplasia seen in partial monosomy 10p such a new risk locus is important because of the patients (Lichtner et al., 2002). severity of AD (Wijsman et al., 2011). References Heart disease Lu X, Timchenko NA, Timchenko LT. Cardiac elav-type Note RNA-binding protein (ETR-3) binds to RNA CUG repeats Arrhythmogenic right ventricular dysplasia is the expanded in myotonic dystrophy. Hum Mol Genet. 1999 most common cause of sudden cardiac death in the Jan;8(1):53-60 young in Italy and the second most common cause Anant S, Henderson JO, Mukhopadhyay D, Navaratnam in the United States. One of the genes that was N, Kennedy S, Min J, Davidson NO. Novel role for RNA- mapped to this is in the vicinity of chromosome binding protein CUGBP2 in mammalian RNA editing. CUGBP2 modulates C to U editing of apolipoprotein B 10p12-p14 and it is CELF2 (Li et al., 2001). mRNA by interacting with apobec-1 and ACF, the apobec- Ischemia 1 complementation factor. J Biol Chem. 2001 Dec 14;276(50):47338-51 Note Ladd AN, Charlet N, Cooper TA. The CELF family of RNA The transient global ischemia induces the binding proteins is implicated in cell-specific and translational inhibition of genes with increased developmentally regulated alternative splicing. Mol Cell expression in normothermic mice. The author's Biol. 2001 Feb;21(4):1285-96 correlate the translational inhibition with CELF2 Li D, Bachinski LL, Roberts R. Genomic organization and expression and this might play an important role in isoform-specific tissue expression of human NAPOR the progress of neuronal injury after transient global (CUGBP2) as a candidate gene for familial arrhythmogenic right ventricular dysplasia. Genomics. 2001 Jun ischemia (Otsuka et al., 2009). 15;74(3):396-401 Atrophy Lichtner P, Attié-Bitach T, Schuffenhauer S, Henwood J, Note Bouvagnet P, Scambler PJ, Meitinger T, Vekemans M. Expression and mutation analysis of BRUNOL3, a The differential expression of CELF2 has been candidate gene for heart and thymus developmental confirmed with real-time RT-PCR in spinal cord defects associated with partial monosomy 10p. J Mol Med and muscle of three different models of spinal (Berl). 2002 Jul;80(7):431-42

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 95 CELF2 (CUGBP, Elav-like family member 2) Ramalingam S, Anant S

Choi DK, Yoo KW, Hong SK, Rhee M, Sakaki Y, Kim CH. growth factor-induced stabilization of cyclooxygenase 2 Isolation and expression of Napor/CUG-BP2 in embryo mRNA in rat smooth muscle cells requires the c-Src family development. Biochem Biophys Res Commun. 2003 Jun of protein-tyrosine kinases. J Biol Chem. 2007 Nov 6;305(3):448-54 9;282(45):32699-709 Gromak N, Matlin AJ, Cooper TA, Smith CW. Antagonistic Ramalingam S, Natarajan G, Schafer C, Subramaniam D, regulation of alpha-actinin alternative splicing by CELF May R, Ramachandran I, Queimado L, Houchen CW, proteins and polypyrimidine tract binding protein. RNA. Anant S. Novel intestinal splice variants of RNA-binding 2003 Apr;9(4):443-56 protein CUGBP2: isoform-specific effects on mitotic catastrophe. Am J Physiol Gastrointest Liver Physiol. 2008 Mukhopadhyay D, Houchen CW, Kennedy S, Dieckgraefe Apr;294(4):G971-81 BK, Anant S. Coupled mRNA stabilization and translational silencing of cyclooxygenase-2 by a novel RNA binding Otsuka N, Tsuritani K, Sakurai T, Kato K, Matoba R, Itoh J, protein, CUGBP2. Mol Cell. 2003a Jan;11(1):113-26 Okuyama S, Yamada K, Yoneda Y. Transcriptional induction and translational inhibition of Arc and Cugbp2 in Mukhopadhyay D, Jung J, Murmu N, Houchen CW, mice hippocampus after transient global ischemia under Dieckgraefe BK, Anant S. CUGBP2 plays a critical role in normothermic condition. Brain Res. 2009 Sep 1;1287:136- apoptosis of breast cancer cells in response to genotoxic 45 injury. Ann N Y Acad Sci. 2003b Dec;1010:504-9 Subramaniam D, Ramalingam S, Linehan DC, Dieckgraefe Anderson KN, Baban D, Oliver PL, Potter A, Davies KE. BK, Postier RG, Houchen CW, Jensen RA, Anant S. RNA Expression profiling in spinal muscular atrophy reveals an binding protein CUGBP2/CELF2 mediates curcumin- RNA binding protein deficit. Neuromuscul Disord. 2004 induced mitotic catastrophe of pancreatic cancer cells. Nov;14(11):711-22 PLoS One. 2011 Feb 11;6(2):e16958 Ladd AN, Cooper TA. Multiple domains control the Wijsman EM, Pankratz ND, Choi Y, Rothstein JH, Faber subcellular localization and activity of ETR-3, a regulator of KM, Cheng R, Lee JH, Bird TD, Bennett DA, Diaz-Arrastia nuclear and cytoplasmic RNA processing events. J Cell R, Goate AM, Farlow M, Ghetti B, Sweet RA, Foroud TM, Sci. 2004 Jul 15;117(Pt 16):3519-29 Mayeux R. Genome-wide association of familial late-onset Murmu N, Jung J, Mukhopadhyay D, Houchen CW, Riehl Alzheimer's disease replicates BIN1 and CLU and TE, Stenson WF, Morrison AR, Arumugam T, Dieckgraefe nominates CUGBP2 in interaction with APOE. PLoS BK, Anant S. Dynamic antagonism between RNA-binding Genet. 2011 Feb;7(2):e1001308 protein CUGBP2 and cyclooxygenase-2-mediated Miyazaki Y, Adachi H, Katsuno M, Minamiyama M, Jiang prostaglandin E2 in radiation damage. Proc Natl Acad Sci YM, Huang Z, Doi H, Matsumoto S, Kondo N, Iida M, U S A. 2004 Sep 21;101(38):13873-8 Tohnai G, Tanaka F, Muramatsu S, Sobue G. Viral Faustino NA, Cooper TA. Identification of putative new delivery of miR-196a ameliorates the SBMA phenotype via splicing targets for ETR-3 using sequences identified by the silencing of CELF2. Nat Med. 2012 Jul;18(7):1136-41 systematic evolution of ligands by exponential enrichment. Ramalingam S, Ramamoorthy P, Subramaniam D, Anant Mol Cell Biol. 2005 Feb;25(3):879-87 S. Reduced Expression of RNA Binding Protein CELF2, a Sureban SM, Murmu N, Rodriguez P, May R, Maheshwari Putative Tumor Suppressor Gene in Colon Cancer. R, Dieckgraefe BK, Houchen CW, Anant S. Functional Immunogastroenterology. 2012;1(1):27-33 antagonism between RNA binding proteins HuR and CUGBP2 determines the fate of COX-2 mRNA translation. This article should be referenced as such: Gastroenterology. 2007 Mar;132(3):1055-65 Ramalingam S, Anant S. CELF2 (CUGBP, Elav-like family Xu K, Kitchen CM, Shu HK, Murphy TJ. Platelet-derived member 2). Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2):93-96.

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CXCL17 (chemokine (C-X-C motif) ligand 17) Aya Matsui, Takashi Murakami Department of Anatomy and Developmental Biology, Graduate School of Medicine, Tokyo Women's Medical University, Tokyo, Japan (AM), Laboratory of Tumor Biology, Faculty of Pharmacy, Takasaki University of Health and Welfare, Gunma, Japan (TM)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/CXCL17ID47679ch19q13.html DOI: 10.4267/2042/56293 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

2006). No Northern blotting signal is detected from Abstract the brain, heart, colon, thymus, spleen, kidney, Review on CXCL17, with data on DNA/RNA, on liver, small intestine, placenta, and peripheral blood the protein encoded and where the gene is lymphocytes (Weinstein et al., 2006). Furthermore, implicated. investigation of CXCL17 expression using a comprehensive human gene expression microarray Identity database showed that expression is restricted to mucosal sites including the digestive system, lung Other names: DMC, Dcip1, UNQ473, VCC-1, airways, the urethra, and several sites of the female VCC1 reproductive system (Burkhardt et al., 2012). HGNC (Hugo): CXCL17 In mouse tissues, CXCL17 is expressed in the lung, Location: 19q13.2 thyroid, submaxillary gland, epididymis and uterus, Note with faint signals in the ovary and prostate CXCL17 was identified as a latest member of the (Weinstein et al., 2006). C-X-C chemokine family in 2006 (Weinstein et al., In rat tissues, real-time PCR using 22 different rat 2006; Pisabarro et al., 2006). Then, it was initially tissues demonstrated that CXCL17 was expressed referred to by other names such as VEGF correlated mainly in the stomach, duodenum, lung, and chemokine 1 (VCC-1) (Weinstein et al., 2006) and salivary gland (Lee et al., 2013). dendritic and monocyte chemokine-like protein (DMC) (Pisabarro et al., 2006). VCC-1 and DMC Protein were identified by cDNA microarray analysis and Description by structure-based protein analysis, respectively. Human CXCL17 has a molecular mass of about DNA/RNA 13.8 kDa, and is a basic protein comprising 119 amino acids. Mouse CXCL17 has a molecular mass Note of about 13.6 kDa, and is also a basic protein The CXCL17 gene is located on the long arm of comprising 119 amino acids. chromosome 19, and it spans about 15 kb and Expression includes four exons. Extensive analyses of CXCL17-secreting or - Description producing cells have yet to be performed. Some Expression of the CXCL17 gene has been observed reports have demonstrated CXCL17 expression by in several tissues. In human tissues, Northern blot immunostaining. Immunohistochemistry of the analysis showed that CXCL17 is expressed in the normal lung in adult humans demonstrated that trachea, stomach, lung, skeletal muscle, and the CXCL17 was constitutively expressed on the fetal lung (Pisabarro et al., 2006; Weinstein et al., bronchial and bronchiolar epithelium.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 97 CXCL17 (chemokine (C-X-C motif) ligand 17) Matsui A, Murakami T

The CXCL17 gene (NM_198477.1). Full-length CXCL17 comprises about 15 kb and includes four exons and three introns. Squares: exon region; two-way arrow: intron region; green: coding region; yellow: untranslated region.

Additionally, CXCL17 expression was also yet to be identified. observed in the normal small intestine (duodenum) and colon. Implicated in Specifically, CXCL17 production was localized to the villus and some crypt epithelial cells of the Breast cancer small intestine and colonic epithelial cells Note (Pisabarro et al., 2006). Immunohistochemical analyses of clinical In mucosal tissues of the gastrointestinal tract, specimens showed that approximately 60% of cells distinct cells were found that exhibited CXCL17- were CXCL17-positive in human breast cancer positivity with unique cell surface markers and (Matsui et al., 2012). morphological characteristics, and these cells were Five out of seven mammary tumors showed + + identified as CD68 macrophages and CD138 considerable CXCL17 up-regulation (from 3-fold to plasma cells. more than 24-fold), in comparison with normal Function mammary tissues (Weinstein et al., 2006). RT-PCR analysis using 13 human breast cancer cell Chemokines are generally classified into one of lines showed CXCL17 expression in 8 cell lines: four groups on the basis of protein structure MDA-MB-361, MCF7, BT-20, BT-474, HCC- (cysteine residues), and can also be classified on the 1419, HCC-1500, HCC-1937 and HCC-1954 basis of functional differences (Zlotnik and Yoshie, (Matsui et al., 2012). 2012). Chemokine groups based on functional Hepatocellular carcinoma characteristics include inflammatory, homeostatic, Note dual-function, plasma or precursor, and platelet CXCL17 is expressed in hepatocellular carcinoma chemokines. (HCC) in humans, but is not expressed in normal However, the functional and physiological role of liver tissues. CXCL17 remains unclear and has yet to be The average rate for CXCL17-positive HCC was categorized. 83% (124 out of 148 samples) in an Interestingly, C-X-C chemokines are divided into immunohistochemical study using clinical HCC two types on the basis of angiogenic potential: pro- specimens (Zhou et al., 2012). angiogenic types with an ELR motif and angiostatic CXCL17 expression in HCC was much higher than types with a non-ELR motif. that in adjacent non-cancerous tissue, and increased The ELR motif consists of the tripeptide glutamate levels of CXCL17 expression were also shown in (E)-leucine (L)-arginine (R). Pro-angiogenic types colon, gastric, breast, lung, bladder, and uterine with an ELR motif which attract neutrophils are cervical cancers (Zhou et al., 2012). referred to as ELR + chemokines, while angiostatic In HCC cell lines, CXCL17-overexpressing HepG2 types that preferentially attract lymphocytes are cells showed increased cell proliferation compared referred to as ELR - chemokines (Vandercappellen to non-expressing control cells (Zhou et al., 2012). et al., 2008; Strieter et al., 2005). The SMMC7721 HCC cell line also showed similar In the case of CXCL17, the presence of the ELR results when CXCL17 was overexpressed (Mu et motif has yet to be confirmed. However, CXCL17 al., 2009). can be functionally classified as an ELR + Moreover, colony formation, invasion and adhesion chemokine since CXCL17 shows pro-angiogenic were promoted following forced expression of function and neutrophil migration (Weinstein et al., CXCL17 (Mu et al., 2009). 2006; Vandercappellen et al., 2008; Zlotnik and When these cells were xenogenically transplanted Yoshie, 2012). into immunodeficient nude mice, tumor growth was Although the function and role of CXCL17 has significantly promoted in comparison with the been extensively investigated, its main receptor has control expression group.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 98 CXCL17 (chemokine (C-X-C motif) ligand 17) Matsui A, Murakami T

Interestingly, whereas control HCC cells subjected However, it has also been reported that CXCL17 to cisplatin treatment (100 µM) induced apoptotic expression in human pancreatic cancer was much morphological changes within 12 hours (e.g., cell higher than in adjacent non-cancerous tissue (Zhou budding, chromatin condensation, nuclear et al., 2012). shrinkage, and cellular fragmentation), CXCL17- In human pancreatic cancer cell lines, two out of expressing HCC cells resisted cisplatin-induced five cell lines (KP-2 and BxPC-3) showed CXCL17 apoptotic cell death (Zhou et al., 2012). mRNA expression (Matsui et al., 2012). Colorectal cancer Fibrosarcoma Note Note In critical specimens, expression of CXCL17 was When mouse fibrosarcoma cells stably transfected 50%-positive in human colorectal cancer (Matsui et with murine CXCL17 cDNA were injected al., 2012; Weinstein et al., 2006). In human subcutaneously into mice, these animals showed colorectal cancer cell lines, four out of nine cell tumor growth retardation compared to parental cell lines (HT-29, KM12, LoVo and COLO205) showed transplantation (Hiraoka et al., 2011). strong expression of CXCL17 mRNA (Matsui et It was subsequently demonstrated that abundant + + + + + al., 2012). In a mice model, xenotransplantation of CD3 , CD4 and CD8 T cells and CD11b CD11c CXCL17 overexpressing DLD-1 and SW620 cells dendritic cells infiltrated CXCL17-expressing into SCID mice resulted in enhanced tumor tumors, suggesting that immune reactions were formation and an increase in the number of evoked in this animal model. intratumoral vessels (Matsui et al., 2012). These Lung cancer tumors were associated with infiltrating CD11b +Gr- 1high neutrophil-like myeloid-derived cells. The Note neutrophil-like myeloid-derived cells were recruited Immunohistochemistry showed that about 30% of at tumor sites by CXCL17. Indeed, SW620 cells non-small cell lung carcinoma was CXCL17- transplanted with CXCL17-responding CD11b +Gr- positive (Matsui et al., 2012). 1high cells resulted in significantly enhanced tumor RT-PCR analysis also demonstrated that CXCL17 formation and angiogenesis compared to control mRNA expression was observed in three out of cells. Furthermore, analyses of metastatic potential eight human lung cancer cell lines (NCI-H441, using in vivo luminescent imaging demonstrated NCI-H1975 and NCI-H2228) (Matsui et al., 2012). that CXCL17-positive cells such as HT-29, KM12 Other tumors and COLO205 were hematogenous distant Note metastases, whereas CXCL17-negative cells such as DLD-1, SW620 and HCT-15 did not show any In RT-PCR analyses of human cancer cell lines, distant metastases (Matsui et al., 2012). CXCL17 expression was demonstrated in one out of six renal cancer cell lines (RTK-2) and in two Endometrial carcinoma out of seven gastric cancer cell lines (MKN-45 and Note KATO III) (Matsui et al., 2012). Gene expression profiling using type I endometrial Human melanoma cell lines investigated were all carcinoma patients demonstrated that 621 out of CXCL17-negative (Matsui et al., 2012). 28869 genes were differentially expressed between Tumorigenicity tumor and normal tissues. Among these 621 genes, 146 were up-regulated and 476 were down- Note regulated in the tumor. One of the up-regulated Tumorigenicity was examined in NIH3T3 cells that genes was CXCL17, although additional biological expressed CXCL17 cDNA. CXCL17- analyses were not reported (Saghir et al., 2010). overexpressing NIH3T3 cells were reported to form tumors in immunodeficient mice (Weinstein et al., Pancreatic cancer 2006; Matsui et al., 2012). Note However, CXCL17 expression itself did not induce In gene expression analyses using different stages focus formation and anchorage-independent cell of pancreatic cancers (intraductal papillary growth using NIH3T3 cells and so CXCL17 lacked mucinous adenoma (IPMA) and intraductal oncogenic transformation activity in vitro (Matsui papillary mucinous carcinoma (IPMC)), CXCL17 et al., 2012). expression was higher in IPMA compared to IPMC Angiogenesis and normal ductal tissues (Hiraoka et al., 2011). In other words, this report suggested that expression of Note CXCL17 was induced at the early stage of In a two-dimensional assay using the mouse carcinogenesis, and then decreased as angioma endothelial cell line PY4.1, vascular tube carcinogenesis progressed. formation in PY4.1 increased CXCL17 expression

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 99 CXCL17 (chemokine (C-X-C motif) ligand 17) Matsui A, Murakami T

levels 28-fold. Adenoviral overexpression of was demonstrated when cells pretreated with CXCL17 in human-derived vascular endothelial CXCL17 showed decreased inflammatory cells (HUVECs) resulted in up-regulation of responses (Lee et al., 2013). J774 murine vascular endothelial growth factor A (VEGF-A) macrophage-like cells immediately express pro- (Weinstein et al., 2006). In another report, inflammatory genes such as IL-6, TNF α and iNOS HUVECs exposed to recombinant CXCL17 when cells are stimulated with LPS. These resulted in increased VEGF-A expression and cell cytokines mediate strong inflammatory responses. migration (Matsui et al., 2012). Investigations of However, pretreatment with CXCL17 significantly the human acute monocytic leukemia cell line THP- reduces LPS-induced expression of pro- 1 showed that CXCL17-treated THP-1 cells inflammatory genes, and LPS-mediated resulted in increased levels of VEGF-A in the inflammatory responses are attenuated. conditioned medium (Lee et al., 2013). Others Immunity Note Note In a microarray-based whole-genome screening CXCL17 has the ability to induce cell migration system using duodenal mucosa during acute (chemotaxis) as is the case with chemokines. It has cholera, expression of CXCL17 was up-regulated in been demonstrated that some immune cells exhibit cholera patients (Flach et al., 2007). In these acute chemotactic cell migration through CXCL17. For cholera patients, mucosal CD8 + cells in the small example, in a transwell migration assay using intestine were localized in the lamina propria human peripheral blood mononuclear cells region. In contrast, these cells migrated from the (PBMCs), CXCL17 specifically induced migration lamina propria region to the epithelium in the of CD14 + monocytes and CD11c + immature convalescent stage. Details concerning the dendritic cells, but not of CD3 + T cells, CD19 + B relationship between CXCL17 expression and cells, CD56 + natural killer cells or CD16 + changes in the localization pattern of the mucosal neutrophils (Pisabarro et al., 2006; Hiraoka et al., CD8 + cells remain to be determined. CXCL17 was 2011). While CD14 + human monocyte-derived produced in bronchoalveolar lavage fluids in mature dendritic cells possessed CXCL17-induced patients with human idiopathic pulmonary fibrosis chemotactic activity, immature Langerhans cells (IPF) (Burkhardt et al., 2012), suggesting that failed to acquire CXCL17-dependent chemotactic CXCL17 production may be associated with human activity (Hiraoka et al., 2011). Furthermore, pulmonary diseases. chemotaxis assays and flow cytometric analyses using murine splenocytes showed that CD11b +Gr- References 1high F4/80 - cells are specifically migrated in a manner dependent on CXCL17 (Matsui et al., Strieter RM, Burdick MD, Gomperts BN, Belperio JA, Keane MP. CXC chemokines in angiogenesis. Cytokine 2012). CXCL17-responding cells showed Growth Factor Rev. 2005 Dec;16(6):593-609 neutrophil-like myeloid-derived cell morphology and cell surface markers. Pisabarro MT, Leung B, Kwong M, Corpuz R, Frantz GD, Chiang N, Vandlen R, Diehl LJ, Skelton N, Kim HS, Eaton Inflammation D, Schmidt KN. Cutting edge: novel human dendritic cell- and monocyte-attracting chemokine-like protein identified Note by fold recognition methods. J Immunol. 2006 Feb Chemokines can control the migration and 15;176(4):2069-73 localization of various cells in the body. However, Weinstein EJ, Head R, Griggs DW, Sun D, Evans RJ, some chemokines exhibit additional activities Swearingen ML, Westlin MM, Mazzarella R. VCC-1, a beyond the trafficking of cells. To date, CXCL17 novel chemokine, promotes tumor growth. Biochem has also been found to possess antimicrobial and Biophys Res Commun. 2006 Nov 10;350(1):74-81 anti-inflammatory activity. The antimicrobial Flach CF, Qadri F, Bhuiyan TR, Alam NH, Jennische E, activity of CXCL17 was demonstrated using Lönnroth I, Holmgren J. Broad up-regulation of innate defense factors during acute cholera. Infect Immun. 2007 Escherichia coli, Staphylococcus aureus, May;75(5):2343-50 Salmonella enterica serovar Typhimurium 14028s, Lactobacillus casei, and Pseudomonas aeruginosa Vandercappellen J, Van Damme J, Struyf S. The role of CXC chemokines and their receptors in cancer. Cancer (Burkhardt et al., 2012). This report demonstrated Lett. 2008 Aug 28;267(2):226-44 that CXCL17 possessed antimicrobial properties, Mu X, Chen Y, Wang S, Huang X, Pan H, Li M. unlike the case with CXCL8 (a representative Overexpression of VCC-1 gene in human hepatocellular chemokine that lacks antimicrobial activity). carcinoma cells promotes cell proliferation and invasion. Moreover, the antimicrobial mechanism by which Acta Biochim Biophys Sin (Shanghai). 2009 CXCL17 induces permeabilization of bacterial Aug;41(8):631-7 membranes was also demonstrated. On the other Saghir FS, Rose IM, Dali AZ, Shamsuddin Z, Jamal AR, hand, the anti-inflammatory activity of CXCL17 Mokhtar NM. Gene expression profiling and cancer-related

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 100 CXCL17 (chemokine (C-X-C motif) ligand 17) Matsui A, Murakami T

pathways in type I endometrial carcinoma. Int J Gynecol high F4/80- cells and promotes tumor progression. PLoS Cancer. 2010 Jul;20(5):724-31 One. 2012;7(8):e44080 Hiraoka N, Yamazaki-Itoh R, Ino Y, Mizuguchi Y, Yamada Zhou Z, Lu X, Zhu P, Zhu W, Mu X, Qu R, Li M. VCC-1 T, Hirohashi S, Kanai Y. CXCL17 and ICAM2 are over-expression inhibits cisplatin-induced apoptosis in associated with a potential anti-tumor immune response in HepG2 cells. Biochem Biophys Res Commun. 2012 Apr early intraepithelial stages of human pancreatic 6;420(2):336-42 carcinogenesis. Gastroenterology. 2011 Jan;140(1):310-21 Zlotnik A, Yoshie O. The chemokine superfamily revisited. Burkhardt AM, Tai KP, Flores-Guiterrez JP, Vilches- Immunity. 2012 May 25;36(5):705-16 Cisneros N, Kamdar K, Barbosa-Quintana O, Valle-Rios R, Hevezi PA, Zuñiga J, Selman M, Ouellette AJ, Zlotnik A. Lee WY, Wang CJ, Lin TY, Hsiao CL, Luo CW. CXCL17, CXCL17 is a mucosal chemokine elevated in idiopathic an orphan chemokine, acts as a novel angiogenic and anti- pulmonary fibrosis that exhibits broad antimicrobial activity. inflammatory factor. Am J Physiol Endocrinol Metab. 2013 J Immunol. 2012 Jun 15;188(12):6399-406 Jan 1;304(1):E32-40 Matsui A, Yokoo H, Negishi Y, Endo-Takahashi Y, Chun This article should be referenced as such: NA, Kadouchi I, Suzuki R, Maruyama K, Aramaki Y, Semba K, Kobayashi E, Takahashi M, Murakami T. Matsui A, Murakami T. CXCL17 (chemokine (C-X-C motif) CXCL17 expression by tumor cells recruits CD11b+Gr1 ligand 17). Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2):97-101.

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Gene Section Short Communication

LRIG3 (leucine-rich repeats and immunoglobulin-like domains 3) Dongsheng Guo, Feng Mao, Ting Lei Dept of Neurosurgery,Tongji Hospital, Huazhong University of Sciences and Technology, Wuhan, Hubei, 430030, People's Republic of China (DG, FM, TL)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/LRIG3ID47529ch12q14.html DOI: 10.4267/2042/56294 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

and 58920538 and spans across 48.3 kilobases. Abstract LRIG3 gene is encoded on the reverse strand. LRIG3 is a member of mammalian LRIG family Transcription and less is known about the functions of LRIG3. Accumulating evidences showed that LRIG3 may Homo sapiens leucine-rich repeats and also function as a tumor suppressor like LRIG1. immunoglobulin-like domains 3 has at least two LRIG3 can inhibit tumor cell proliferation, survival, transcript variants. The pre-mRNAs comprise 19 adhesion and invasion in many types of human exons. The coding sequence of transcript variant 1 cancer cells based on cell experiments. Perinuclear (NM_001136051.2) and 2 (NM_153377.4) is staining of LRIG3 correlated with better survival in 3691bp and 4110bp respectively. astrocytic tumors. Decreased LRIG3 was identified as one of 12 promising serum biomarkers for early Protein stage non-small-cell lung cancer. LRIG3 may play Description a role in inner ear morphogenesis and neural crest formation. LRIG3 is a transmembrane cell-surface protein consisting of 1059 or 1119 amino acids. LRIG3 Keywords contains a signal peptide, 15 tandem LRR with LRIG3, glioma, ependymoma, NSCLC, pituitary cysteine rich N- and C-flanking domains, three adenoma, cervical adenocarcinoma, psoriasis immunoglobulin-like domains, a transmembrane Identity domain, and a cytoplasmic tail. Expression Other names: LIG3 LRIG3 is ubiquitously expressed in human organs. HGNC (Hugo): LRIG3 The highest expression of LRIG3 is in stomach, Location: 12q14.1 thyroid, and skin. Localisation DNA/RNA Differential subcellular expression in a cell type- Description sepcific manner. According to Entrez-Gene, LRIG3 gene maps to Function NC_000012.12 in the region between 58872155 LRIG3 may function as a tumor suppressor.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 102 LRIG3 (leucine-rich repeats and immunoglobulin-like domains Guo D, et al. 3)

LRIG3 protein. SP: signal peptide; NF: N-terminal cysteine-rich flanking domain; LRRs: 15 leucine-rich repeats; CF: C-terminal cysteinerich flanking domain; Ig C2: C2-type immunoglobulin-like domains; TM: transmembrane domain; Cyto: cytoplasmic domain.

Mutations Psoriasis Note Note Mutations in LRIG3 are rare, according to the The expression pattern of LRIG3 protein was TCGA data set. altered in psoriatic lesions compared to normal skin (Karlsson et al., 2008). Implicated in Inner ear morphogenesis Ependymoma Note During inner ear development, Lrig3 acts to repress Note Netrin transcription and Lrig3 loss led to defects in Perinuclear staining of LRIG3 was significantly inner ear morphogenesis (Abraira et al., 2008). lower in posterior fossa ependymomas than in supratentorial ependymomas (Yi et al., 2009). Neural crest formation Pituitary adenoma Note LRIG3 is required for neural crest formation in Note developing Xenopus embryos by regulating the The expression of LRIG 3 in HP75 cell line was Wnt and Fgf signaling pathways (Zhao et al., lower compared to normal pituitary glands (Guo et 2008). al., 2007). Astrocytic tumors To be noted Note Note Perinuclear expression of LRIG3 in astrocytic Recently, Colleen Sweeney and Lisa V Goodrich tumors has been associated with better patient reported that overexpression of LRIG3 in survival (Guo et al., 2006). HEK293T cells increased the ERBB receptor Down-regulation of LRIG3 expression in expression (Rafidi et al., 2013). However, glioblastoma cell lines resulted in increased downregulation of LRIG3 in a human glioblastoma proliferation and invasion, decreased apoptosis, and cell line, GL15, resulted in upregulation of total increased EGFR expression (Cai et al., 2009). EGFR and phospho-EGFR, and enchanced glioma Cervical adenocarcinoma cell proliferation, adhesion and invasion (Cai et al., 2009). Note Therefore, the precise function of LRIG3 in regard High fraction of LRIG3-positive cells were to ERBB receptor signalling modulation may be significantly associated with patient survival, and context-dependent and the role of LRIG3 needs to LRIG3 staining intensity was positively correlated be further studied. with HPV status (Muller et al., 2013). Non-small-cell lung cancer References Note Guo D, Holmlund C, Henriksson R, Hedman H. The LRIG gene family has three vertebrate paralogs widely LRIG3 downregulation was identified as one of 12 expressed in human and mouse tissues and a homolog in promising serum biomarkers for early stage non- Ascidiacea. Genomics. 2004 Jul;84(1):157-65 small-cell lung cancer (Ostroff et al., 2010). Guo D, Nilsson J, Haapasalo H, Raheem O, Bergenheim Abnormal protein T, Hedman H, Henriksson R. Perinuclear leucine-rich The LRIG3-ROS1 fusion has been identified in repeats and immunoglobulin-like domain proteins (LRIG1- 3) as prognostic indicators in astrocytic tumors. Acta non-small-cell lung cancer (Cai et al., 2013). Neuropathol. 2006 Mar;111(3):238-46

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 103 LRIG3 (leucine-rich repeats and immunoglobulin-like domains Guo D, et al. 3)

Guo D, Han L, Shu K, Chen J, Lei T. Down-regulation of Ghasimi S, Haapasalo H, Eray M, Korhonen K, leucine-rich repeats and immunoglobulin-like domain Brännström T, Hedman H, Andersson U. proteins (LRIG1-3) in HP75 pituitary adenoma cell line. J Immunohistochemical analysis of LRIG proteins in Huazhong Univ Sci Technolog Med Sci. 2007 meningiomas: correlation between estrogen receptor Feb;27(1):91-4 status and LRIG expression. J Neurooncol. 2012 Jul;108(3):435-41 Abraira VE, Del Rio T, Tucker AF, Slonimsky J, Keirnes HL, Goodrich LV. Cross-repressive interactions between Teo AK, Ali Y, Wong KY, Chipperfield H, Sadasivam A, Lrig3 and netrin 1 shape the architecture of the inner ear. Poobalan Y, Tan EK, Wang ST, Abraham S, Tsuneyoshi Development. 2008 Dec;135(24):4091-9 N, Stanton LW, Dunn NR. Activin and BMP4 synergistically promote formation of definitive endoderm in human Karlsson T, Mark EB, Henriksson R, Hedman H. embryonic stem cells. Stem Cells. 2012 Apr;30(4):631-42 Redistribution of LRIG proteins in psoriasis. J Invest Dermatol. 2008 May;128(5):1192-5 Wu X, Hedman H, Bergqvist M, Bergström S, Henriksson R, Gullbo J, Lennartsson J, Hesselius P, Ekman S. Zhao H, Tanegashima K, Ro H, Dawid IB. Lrig3 regulates Expression of EGFR and LRIG proteins in oesophageal neural crest formation in Xenopus by modulating Fgf and carcinoma with emphasis on patient survival and cellular Wnt signaling pathways. Development. 2008 chemosensitivity. Acta Oncol. 2012 Jan;51(1):69-76 Apr;135(7):1283-93 Cai W, Li X, Su C, Fan L, Zheng L, Fei K, Zhou C, Cai M, Xie R, Han L, Chen R, Wang B, Ye F, Guo D, Lei T. Manegold C, Schmid-Bindert G. ROS1 fusions in Chinese Effects of RNAi-mediated gene silencing of LRIG3 patients with non-small-cell lung cancer. Ann Oncol. 2013 expression on cell cycle and survival of glioma cells. J Jul;24(7):1822-7 Huazhong Univ Sci Technolog Med Sci. 2009 Feb;29(1):88-93 Del Rio T, Nishitani AM, Yu WM, Goodrich LV. In vivo analysis of Lrig genes reveals redundant and independent Cai M, Han L, Chen R, Ye F, Wang B, Han F, Lei T, Guo functions in the inner ear. PLoS Genet. D. Inhibition of LRIG3 gene expression via RNA 2013;9(9):e1003824 interference modulates the proliferation, cell cycle, cell apoptosis, adhesion and invasion of glioblastoma cell Muller S, Lindquist D, Kanter L, Flores-Staino C, (GL15). Cancer Lett. 2009 Jun 8;278(1):104-12 Henriksson R, Hedman H, Andersson S. Expression of LRIG1 and LRIG3 correlates with human papillomavirus Yi W, Haapasalo H, Holmlund C, Järvelä S, Raheem O, status and patient survival in cervical adenocarcinoma. Int Bergenheim AT, Hedman H, Henriksson R. Expression of J Oncol. 2013 Jan;42(1):247-52 leucine-rich repeats and immunoglobulin-like domains (LRIG) proteins in human ependymoma relates to tumor Qi Y, Chang L, Li H, Yu G, Xiao W, Xia D, Guan W, Yang location, WHO grade, and patient age. Clin Neuropathol. Y, Lang B, Deng KL, Yao WM, Ye ZQ, Zhuang QY. Over- 2009 Jan-Feb;28(1):21-7 expression of LRIG3 suppresses growth and invasion of bladder cancer cells. J Huazhong Univ Sci Technolog Med Abraira VE, Satoh T, Fekete DM, Goodrich LV. Vertebrate Sci. 2013 Feb;33(1):111-6 Lrig3-ErbB interactions occur in vitro but are unlikely to play a role in Lrig3-dependent inner ear morphogenesis. Rafidi H, Mercado F 3rd, Astudillo M, Fry WH, Saldana M, PLoS One. 2010 Feb 1;5(2):e8981 Carraway KL 3rd, Sweeney C. Leucine-rich repeat and immunoglobulin domain-containing protein-1 (Lrig1) Ostroff RM, Bigbee WL, Franklin W, Gold L, Mehan M, negative regulatory action toward ErbB receptor tyrosine Miller YE, Pass HI, Rom WN, Siegfried JM, Stewart A, kinases is opposed by leucine-rich repeat and Walker JJ, Weissfeld JL, Williams S, Zichi D, Brody EN. immunoglobulin domain-containing protein 3 (Lrig3). J Biol Unlocking biomarker discovery: large scale application of Chem. 2013 Jul 26;288(30):21593-605 aptamer proteomic technology for early detection of lung cancer. PLoS One. 2010 Dec 7;5(12):e15003 Wang Y, Poulin EJ, Coffey RJ. LRIG1 is a triple threat: ERBB negative regulator, intestinal stem cell marker and Yang H, Mao F, Zhang H, Wang B, Wan F, Guo D, Lei T. tumour suppressor. Br J Cancer. 2013 May Effect of over-expressed LRIG3 on cell cycle and survival 14;108(9):1765-70 of glioma cells. J Huazhong Univ Sci Technolog Med Sci. 2011 Oct;31(5):667-72 This article should be referenced as such: Yuan X, Bao S, Yang W, Ye Z. Effect of silencing LRIG3 Guo D, Mao F, Lei T. LRIG3 (leucine-rich repeats and gene on the proliferation and apoptosis of bladder cancer immunoglobulin-like domains 3). Atlas Genet Cytogenet T24 cells. J Huazhong Univ Sci Technolog Med Sci. 2011 Oncol Haematol. 2015; 19(2):102-104. Apr;31(2):220-5

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Gene Section Review

MKI67 (marker of proliferation Ki-67) Konstantinos Ntzeros, Philip Stanier, Dimitrios Mazis, Neoklis Kritikos, Meletios Rozis, Eustathios Anesidis, Chrisoula Antoniou, Michael Stamatakos Astros Medical Center, Panarcadian General Hospital of Tripoli, Tripoli, Greece (KN), UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK (PS), REA Maternity Hospital, Athens, Greece (DM), Nursing Unit of Molaoi, General Hospital of Lakonia, Molaoi, Greece (NK, EA, MS), Surgery Department, General Hospital of Korinthos, Korinthos, Greece (MR), Obstetrics and Gynecology Department, General Hospital of Nikaia "St. Panteleimon", Piraeus, Greece (CA)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/MKI67ID41369ch10q26.html DOI: 10.4267/2042/56295 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

10q26.2, starting at 128096659 and ending at Abstract 128126204 bp from pter (according to GRCh38 Review on MKI67, with data on DNA/RNA, on the Primary Assembly). The gene size is 29,545 bp protein encoded and where the gene is implicated. consisting of 15 exons and 14 introns. The gene has a minus strand orientation. Identity Description Other names: KIA, MIB-1 Promoter: The promoter of MKI67 is located HGNC (Hugo): MKI67 upstream of the transcription start site. Several Location: 10q26.2 studies have shown that the 5' flanking region of the gene has promoter activity via reporter gene Local order expression assays and deletion analysis (Zambon, CLRN3, PTPRE, MKI67 , LINC01163, MGMT. 2010; Pei et al., 2012). The promoter region was Note shown to contain a TATA-less, GC rich region with MKI67 is the human homologue of the product several putative Sp1 binding sites and two recognized by the monoclonal antibody Ki67, evolutionary conserved E2F transcription factor which is used as a tumor proliferation marker. binding sites (Zambon, 2010; Pei et al., 2012). Gene: The gene consists of 15 exons and 14 DNA/RNA introns. The first exon and a part of the second form the 5' Untranslated Region (5' UTR) of the mRNA Note transcript (Duchrow et al., 1996; Schlüter et al., MKI67 is located on chromosome 10 at locus 1993).

The local order of genes around MKI67. The MKI67 gene is surrounded by the genes CLRN3 (clarin 3), PTPRE (protein tyrosine phospatase receptor type E), MGMT (O-6-methylguanine-DNA methyltransferase).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 105 MKI67 (marker of proliferation Ki-67) Ntzeros K, et al.

MKI67 gene structure. The UTRs are coloured green and the coding regions red. The introns are coloured yellow. MKI67 consists of 15 exons of which the first and a part of the second exon form the 5' UTR. The 3' UTR is formed from a major part of exon 15, which contains only a small coding region. The table at the bottom of the figure shows the length of exons, coding regions and introns according to the information provided by the NM_002417.4 NCBI reference sequence.

The second exon contains the start codon (Schlüter 12497 bp (NCBI Reference Sequence: et al., 1993), with the open reading frame extending NM_002417.4). The "short type" mRNA splice to the 15th exon, which consists of a short coding variant consists of 14 exons lacking the 7th exon sequences, the stop codon and the 3' UTR and its annotated spliced exon length is 11417 bp (Duchrow et al., 1996; Schlüter et al., 1993). (NCBI Reference Sequence: NM_001145966.1). The 1080 bp 7th exon is not present in the alternatively spliced "short type" mRNA transcript. Pseudogene The 6845 bp 13th exon contains the characteristic There is one related pseudogene of MKI67 on sixteen homologous "Ki67 repeats", each of them chromosome X, named MKI67P1 (marker of having length of 366 bp. Within Ki67 repeats there proliferation Ki-67 pseudogene 1) with Gene ID is a highly conserved new motif of 66 bp named the 100271918. The MKI67P1 pseudogene is located "Ki67 motif" (Schlüter et al., 1993). on chromosomal locus Xp11. It has 1449 bp length The introns, vary in size from 87 to 3561 bp. Three and NCBI Reference Sequence: NG_011647.1. introns contain recognized homologue copies of "Alu-repeats". All the intron-exon transitions Protein contain a potential branch site with the exception of the introns flanking exon 7 where there is not any Note donor-acceptor splicing signal (Durchow et al., The MKI67 protein can be found in two complete 1996). isoforms, the heavy isoform produced from the "long type" mRNA and the light isoform translated Transcription from the "short type" mRNA, lacking the 7th exon. There are two mRNA splice variants of MKI67, The heavy isoform, referred as Antigen Ki67 that are shown to be processed through translation. isoform 1 with NCBI Reference Sequence: These two mRNA variants are named as "long NP_002408.3, consists of 3256 aminoacids (aa) and type" and "short type" since they differ only by the weights approximately 395 kDa. The light isoform, presence or absence of exon 7 (Schlüter et al., referred as Antigen Ki67 isoform 2 with NCBI 1993). Reference Sequence: NP_001139438.1, consists of The "long type" mRNA splice variant consists of 15 2896 aa lacking the 360 aa encoded by exon 7 and exons and its total annotated spliced exon length is has a molecular weight of about 345 kDa.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 106 MKI67 (marker of proliferation Ki-67) Ntzeros K, et al.

The MKI67 mRNA and protein. The two types of mRNA transcripts, shown to be translated to protein, are depicted in the first part of the figure. The long type mRNA differs from the short type by the presence of the 7th exon which is alternatively spliced in the short type. The UTRs are coloured green and the coding regions red. At the second part of the figure, it is depicted the protein product of the long type mRNA which contains all the putative domains of this protein. The major protein domains are the FHA domain coloured red, the PPI binding site coloured purple, the Ki67 repeats coloured yellow and the ATP/ GTP binding site motif A-P loop coloured blue. There are other putative functional regions within these major domains, which are highlighted in different colours.

Description aa, 4) 1368-1478 aa, 5) 1489-1599 aa, 6) 1610- 1721 aa, 7) 1732-1843 aa, 8) 1854-1965 aa, 9) Starting from the N-terminus: 1976-2087 aa, 10) 2098-2204 aa, 11) 2216-2327 aa, Major domains: 12) 2337-2448 aa, 13) 2459-2570 aa, 14) 2582- - Forkhead associated domain (FHA): 8-98 aa. 2690 aa, 15) 2701-2806 aa, 16) 2820-2929 aa. This domain is characterized as a modular The Ki67 repeat is a cluster of 122 residues phosphopeptide recognition domain with specificity repeated 16 times within the 13th exon of the to phosphothreonine containing sequences molecule with identity ranging from 43 to 62% to (Hammet et al., 2003; Durocher et al., 1999). The the consensus sequence. MKI67 FHA domain is shown to lie at the N- Within the Ki67 repeat there is a highly conserved terminus of the protein. According to computer motif of 22 amino acids named "the Ki67 motif" alignment, the core domain is found within 27-76 which has 72-100% identity to the consensus amino acids but the functional domain is believed sequence and includes the F-K-E-L epitope to expand from 8 to 98 residue. So far, two proteins recognized by the prototype Ki67 antibody have been identified to interact with MKI67 FHA (Schlüter et al., 1993). domain, the hNIFK (human nucleolar protein So far, their function remains an enigma. interacting with the FHA domain of pKi-67) and - ATP/ GTP binding site motif A-P loop: 3034- the Hklp2 (human kinesin-like protein 2). 3041 aa. - Protein phosphatase 1 binding site: 502-563 aa. A-P-R-A-R-G-K-S The Protein Phosphatase 1 (PPI) is a Ser/Thr At the C-terminus of the protein there is a sequence phosphatase which belongs to the PPP family of resembling the predictive nucleotide binding site of phosphatases, and it is believed to dephosphorylate Walker A motif. its substrates in large complexes consisting of This well-known motif is a nucleotide binding fold regulatory as well as target proteins (Moorhead et recognized by Walker et al. (1982) and it is found al., 2008). In MKI67 protein there is a docking site in many nucleotide binding proteins. of PPI at the N-terminal part of the protein within Minor domains: residues 502 to 563. - PEST sequences: - Ki67 Repeats: The cDNA sequences show 40 weak and 10 strong 1) 1002-1113 aa, 2) 1124-1235 aa, 3) 1246-1357 PEST sites.

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These sites are named as PEST due to the fact that for interacting with the HiNF-P transcription factor they are rich in proline, glutamic acid (E), serine in a yeast two-hybrid screen (Miele et al., 2007). and threonine and to a lesser extent aspartic acid. Finally, several studies have shown that MKI67 They are found in several proteins with diverse protein interacts with DNA and more specifically functions such as key metabolic enzymes, with the heterochromatin (Kreitz et al., 2000; transcription factors, protein kinases, phosphatases Bridger et al., 1998). and cyclins. The regulation of proteins based on Expression PEST sequences is characterized by strict expression, high susceptibility to proteolysis and The expression profile of MKI67 is much more short half-life, especially in those proteins related to the proliferating state of a cell rather than participating in cell cycle and mitosis regulation. In to the histological background of that cell. Through MKI67 protein, the 10 strongest PEST sequences immunostaining experiments Gerdes et al. (1983) are located within exon 13, surrounding the showed that this gene is present in proliferating conserved cysteine residues at position 8 of the normal cells such as germinal centers of cortical Ki67 repeats (Ross and Hall, 1995). PEST follicles, cortical thymocytes, neck cells of sequences seem to be functional in the MKI67 gastrointestinal mucosa and other cell lines but protein, based on its biological behavior with absent in resting differentiated cells such as susceptibility to proteases, short half-life and rapid lymphocytes, monocytes, hepatocytes, renal, brain, loss of the protein after mitosis (Ross and Hall, or parietal cells and many others. 1995; Schlüter et al., 1993). A very important finding was that the expression of - Putative Monopartite Nuclear Targeting MKI67 was triggered in differentiated cells after Sequences: 502-505 and 687-690 aa. proliferative stimulation and disappeared after The MKI67 protein has two putative monopartite differentiation of proliferating cells (Gerdes et al., nuclear targeting sequences at the N-terminal part 1983). of the protein, which could function as nuclear During cell cycle this gene is always detectable localization signals (NLS) through the classical through G1, S, G2 and M phase at continuously importin a/b nuclear import mechanism. proliferating cells but totally absent in resting cells - Putative Bipartite Nuclear Targeting Sequences: at G0 phase (Gerdes et al., 1984). 1) 536-550 aa, 2) 1516-1530 aa, 3) 2244-2258 aa, Localisation 4) 2365-2379 aa, 5) 2651-2665 aa, 6) 2890-2904 Distribution of MKI67 protein during cell cycle: aa, 7) 2997-3011 aa, 8) 3141-3155 aa. 1. Interphase: There are other classical nuclear targeting a. Early G1 phase: Accumulates in several foci sequences within this molecule, which could within the nucleoplasm (Gerdes et al., 1983; Kill, mediate nuclear localization with importin a/b 1996). pathway as well. These bipartite sequences are b. Late G1, S, G2 phase: Localizes predominantly characterized by two clusters of basic residues within nucleoli at the dense fibrillar component but separated by a 10 to 12 aa linker that is tolerant to also there is diffuse nucleoplasmic staining (Gerdes residue substitution (Kosugi et al., 2009). et al., 1983; Kill, 1996). - Post-translational modifications: 2. Mitosis: Coats chromosome surface (Gerdes et The predicted post-translational modifications of al., 1983). MKI67 protein comprise 19 N-myristoylation, 3 a. Metaphase: a reticulate but uniform network of amidation and over 200 phosphorylation sites (143 fibrils around chromosomes, at the PKC, 89 casein kinase II, 2 tyrosine kinase sites perichromosomal layer (Ross and Hall, 1995). and 8 consensus sites for Cdc2 kinase) (Schlüter et b. Anaphase: a reticulate more granular network of al., 1993). fibrils around chromosomes with the highest - MKI67 protein interactions: density of staining (Ross and Hall, 1995). The MKI67 protein has been found to interact c. Telophase: the staining moves from the directly with four proteins. The HP1 proteins, perichromosomal layer and becomes speckled at the which are small non-histone chromosomal proteins nucleoplasm and subsequently concentrates at the found in several chromatin complexes, interact with newly reformed nucleoli (Ross and Hall, 1995). its C-terminal region (Kametaka et al., 2002). The Hklp2, a kinesin-like motor, interacts with the N- Function terminal FHA domain of MKI67 protein (Sueishi et The function of MKI67 protein during cell cycle is al., 2000). Another protein shown to interact with still unknown. However, experiments with anti- the N-terminal FHA domain of MKI67 protein is a sense oligonucleotides and antibodies against putative RNA-binding protein with length of 293 MKI67 revealed a decreased rate of cell division, residues, named hNIFK (Takagi et al., 2001). indicating the important role of this protein in cell MKI67 protein was found to be a candidate protein cycle (Schlüter et al., 1993; Starborg et al., 1996).

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More details for MKI67 function are unavailable, in tumor development is unknown. In 2003, Buban although there are some assumptions based on et al. found eight different point mutations in four localization and protein-protein interactions tumor cell lines (HeLa, CXF94, SW480, A549) experimental data. indicating that MKI67 might provide a genetic Models: background in tumor development. These mutations 1. Nucleolar localization of MKI67 protein is include a deletion in position 1496 resulting in a believed to be related to: truncated product, a base exchange silent mutation a. Structural modulation of the nucleolus in order to in position 433 (A433T) and six other exchange enhance the high rates of ribosomal synthesis mutations resulting in residue changes (Buban et during cell proliferation, since its expression is al., 2004). related to high rates of protein synthesis (Plaat et al., 1999; Scholzen et al., 2002). Implicated in b. Sequestration of MKI67 protein within nucleoli Several cancers until the mitosis starts again. During mitosis the nucleolar structures are dispersed and the MKI67 Note protein is free to interact with nuclear components Although the MKI67 gene is not associated with (Scholzen et al., 2002). any type of human cancer as a causative factor, it is 2. Localization of MKI67 to the surfaces of implicated in many of them as a prognostic factor chromosomes during mitosis (prometaphase to based on the expression profile of this gene at anaphase) is believed to be related to: tumor cells. Additionally, a new application of a. Protection of chromosomal surface during MKI67 expression is the implication in diagnosis of mitosis (Verheijen et al., 1989; Yasuda and Maul, certain diseases such as lymphoma. The prognostic 1990). value of MKI67 expression in malignancy was 3. DNA interaction: categorized by Brown and Gatter (2002) into three a. It was believed that the central part of the MKI67 classes: protein and especially the "Ki67 repeat" region has 1. Group of malignancies where >75% of the the ability to bind DNA through its Thr-Pro-X-X studies shown significant prognostic value: breast motif (Schlüter et al., 1993). cancer, soft tissue tumors, lung cancer, astrocytoma b. The LR domain has the ability to bind DNA in and meningioma. order to compact chromatin (Kametaka et al., 2. Group of malignancies where 25-75% of the 2002). studies shown significant prognostic value: cervical cancer, prostate cancer. Homology 3. Group of malignancies where <25% of the The MKI67 gene has homologous genes in several studies shown significant prognostic value: organisms such as chimpanzee, rhesus monkey, colorectal cancer. dog, cattle, mouse and rat. Additionally, through genome annotation studies, Breast cancer 53 other organisms have been found to carry an Note MKI67 orthologue. However, the most interesting In breast cancer, most of the studies show case is a similar but not homologous gene, found in significant correlation between MKI67 expression the long-nosed rat Kangaroo, named chmadrin in tumor cells and clinical outcomes both in (Takagi et al., 1999). The chmadrin protein has univariate and multivariate analysis (Brown and primary structural similarities with the MKI67 Gatter, 2002; Kontzoglou et al., 2013). The protein. The N-terminal region of the chmadrin prognostic value of MKI67 expression in breast protein has a nucleotide binding motif which is cancer is comprised in the following conclusions: similar to a putative one found in MKI67 at the C- 1. MKI67 expression is associated with common terminal region (Takagi et al., 1999). The central histopathological parameters and the strongest region of chmadrin contains a repetitive sequence correlation is established with tumor grading domain, named the "chmadrin repeat domain" (Inwald et al., 2013). which resembles the "Ki67 repeat domain" (Takagi 2. MKI67 expression is an independent prognostic et al., 1999). Finally, both of these proteins have parameter of overall and disease-free survival LR domains with conserved functionality but (Inwald et al., 2013). different amino acid sequence (Takagi et al., 1999). 3. MKI67 expression is associated with earlier central nervous system metastases (Ishihara et al., Mutations 2013). 4. In triple negative breast cancer (TNBC) MKI67 Note expression could be used to classify TNBC into two MKI67 is used as a tumor proliferation marker with subtypes with different prognosis (Keam et al., an unknown function. The contribution of this gene 2011).

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5. In node-negative breast cancer, which is treated expression rather than MKI67 gene (Li et al., 2014; with surgery and subsequent radiation but not with Salvesen et al., 1999; Gassel et al., 1998). adjuvant systemic therapy, MKI67 expression and Cervical cancer hormone receptor status is a significant prognostic parameter of survival (Pathmanathan et al., 2014). Note 6. The expression of MKI67 gene after neoadjuvant In cervical cancer MKI67 expression is used as a chemotherapy is prognostic of disease relapse and proliferation index marker of malignant cells, death (von Minckwitz et al., 2013; Tanei et al., associated with specific characteristics of the tumor 2011). but not as a prognostic factor of survival or disease 7. The prognostic value of MKI67 expression is relapse. available in samples of breast tumors acquired via Additionally, it is suggested that under certain fine needle aspirate with accuracy comparable to circumstances MKI67 expression could be used in histological evaluation (Konofaos et al., 2013). low-grade lesion triage for referral colposcopy. 8. Breast cancer patients with Grade 3 tumors More specifically: positive for MS110, Lys27H3, VIM (vimentin) and 1. There is no significant correlation between MKI67 expression are at high risk of carrying MKI67 expression and classical prognostic factors BRCA1 mutations and therefore could be screened (Ancuta et al., 2009). for such mutations (Hassanein et al., 2013). 2. MKI67 and CDKN2A (P16) gene is expressed in Additionally, another diagnostic use of MKI67 cervical intraepithelial neoplasia (CIN) and the expression in breast cancer addresses two specific intensity of positive expression is significantly subtypes of this malignancy: correlated with CIN grade (Shi et al., 2007). In 1. In Phyllodes tymor the expression of MKI67 and another study these two markers were shown to TP53 mRNA is associated with the grade of this increase linearly from control cases to more tumor and could help in distinguishing the benign dysplastic lesion to squamous cell carcinoma (Gatta from the malignant form (Kucuk et al., 2013). et al., 2011). This indicates that CDKN2A and 2. The expression profile of the estrogen receptor, MKI67 gene could be useful markers in diagnosis ERBB2 (c-erbB2) and MKI67 mRNA could help in and staging of CIN lesions (Shi et al., 2007). distinguishing between Toker cells, which are 3. Expression of CDKN2A, MKI67 gene and normal components of nipple epidermis, and cells ProxEX C (a cocktail of monoclonal antibodies of Paget's disease (Park and Suh, 2009). against proteins associated with aberrant S phase cell cycle induction) are most associated with the Endometrial cancer severity of cervical dysplasia and related to HPV-16 Note infection (Conesa-Zamora et al., 2009). Although in endometrial cancer MKI67 expression 4. MKI67 expression is enhanced in high grade is used as a proliferation marker associated with CIN lesions and cervical squamous cell carcinoma specific characteristics of this tumor, there is not yet (Looi et al., 2008). TP63 and MKI67 expression is any validated prognostic or diagnostic connection correlated better with cancer progress than TP53 as there is for breast cancer. More specifically: expression (Vasilescu et al., 2009). 1. High MKI67 expression is associated with 5. CDKN2A/MKI67 expression increases endometrial cancer and the overexpression of according to histologic severity and could be used MKI67 and TP53 genes indicates a more malignant to predict high-grade lesion better than HPV DNA phenotype with poor differentiation of the tumor testing since it is more accurate in patients with cells (Markova et al., 2010). atypical squamous cells (Koo et al., 2013). 2. High MKI67 expression is correlated with poorly Additionally, they could be used for low-grade differentiated carcinomas, invasion of the squamous intraepithelial lesions triage better than myometrium and stage III tumor (Stoian et al., HPV DNA testing and reduce referral colposcopy 2011). There is also a significant correlation with to almost the half by detecting the more severe the degree of differentiation, the stage of tumor and cases of CIN3 (Wentzensen et al., 2012). vascular invasion (Stoian et al., 2011). Ovarian cancer 3. MKI67 expression is higher in high-grade endometrial carcinomas and complete negative in Note atrophic endometrium (Mourtzikou et al., 2012). In ovarian cancer MKI67 expression is associated 4. High MKI67 expression correlates with with several histopathological characteristics of the morphologic features of aggressiveness and high malignancy and in some subtypes with the overall grade of endometrial cancer (Konstantinos et al., survival. However, more studies are needed in order 2013). to validate such correlations. More specifically: 5. Although previous studies associated MKI67 1. PIK3CA amplifications and MKI67 expression expression with survival, recent ones correlate are strong predictors of an early tumor-associated survival with estrogen and progesterone receptors death (Woenckhaus et al., 2007).

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2. The combined evaluation of CDKN1B Bladder cancer (P27KIP1) and MKI67 expression in ovarian Note carcinomas is an independent and more accurate In bladder cancer MKI67 expression is examined in prognostic marker of overall survival than each several studies classifying this gene as a common marker alone (Korkolopoulou et al., 2002). marker used in diagnosis and staging of this 3. In stage III ovarian carcinomas, MKI67 malignancy. The expression of this gene is expression is an independent prognostic marker correlated with the grade and the stage of the where high expression of this gene is significantly malignancy and it is higher in females (Wang et al., associated with shorter survival (Khouja et al., 2013). 2007). 4. The expression of MKI67 gene is higher in Gliomas: astrocytomas, malignant ovarian tissues than control ones and oligodendrogliomas within the malignant tissues it is enhanced in low Note differential carcinoma and stage III-IV (Wang et al., In these brain tumors the MKI67 gene has great 2010b). potential as a marker of cell proliferation, survival, 5. The expression of MKI67 gene is higher in therapy response and even discrimination between primary ovarian tumors than in metastatic cells subtypes of gliomas. High expression of this gene is (Wang et al., 2010b). related with shorter overall survival of patients (Liu 6. The high-grade ovarian carcinomas have et al., 2013b). Additionally, MKI67 expression was significantly higher expression of MKI67 gene than shown to be correlated with survival of patients the low-grade, but there is little correlation between with a specific subtype of gliomas in univariable this expression and overall or disease-free survival analysis, indicating the prognostic potential of this (Shen et al., 2011). gene in glioma tumors (Preusser et al., 2012). The 7. The MKI67 expression is associated with higher MKI67 expression could also be associated with cancer stage and histological grade and a worse post-treatment results, such as in adjuvant outcome for patients (Heeran et al., 2013). radiotherapy of gliomas where high expression is 8. The MKI67 expression was found to be correlated with worse response to the therapy associated with poorer overall survival in ovarian (Horbinski et al., 2012). Another interesting study cancer patients (Kucukgoz Gulec et al., 2014). showed that MKI67 might have differential Vulvar cancer expression between subtypes of gliomas indicating a role in diagnosis of these subtypes in accordance Note to other promising molecular markers (Huang et al., The vulvar carcinomas, mainly represented by 2011). squamous cell carcinoma (SCC) and non-invasive lesion of vulvar intraepithelial neoplasias (VINs), Meningiomas are associated with MKI67 expression in several Note studies. MKI67 is believed to have some influence in The general idea is that the expression of this gene meningioma development and progression since is associated with dysplastic lesions and it is more there is significant correlation between its intense in more malignant lesions. More expression and tumor grade, certain subtypes, specifically: tumor recurrence and size (Pavelin et al., 2013; 1. The expression of MKI67 and CDKN2A are Wang et al., 2010a). Another interesting correlation statistically positively related with HPV-associated was proposed by Uzum and Ataoglu (2008) in dysplastic lesions (Gincheva et al., 2009). which the tumor grade is the most important 2. There is an increase in MKI67 and TOP2A prognostic factor of meningiomas, which in turn is (topoisomerase IIa) expression in VIN lesions and significantly correlated with MKI67 expression. SCC comparing to normal vulvar epithelia, Terzi et al. (2008) proposed that MKI67 and TP53 indicating these two genes as markers of expression could be useful additional markers for proliferation in vulvar epithelial tissues (Brustmann grade classification in borderline cases. and Naude, 2002). 3. In differentiated VIN lesions the MKI67 Pituitary adenomas expression is confined in the basal and parabasal Note layers. The MKI67 expression is correlated with neoplasm However, in usual VIN lesions associated with recurrence and visual field defect (Paek et al., high-risk HPV virus infection, MKI67 is highly 2005). Additionally, it is believed to be an expressed (Hoevenaars et al., 2008; Ding et al., independent predictor of pituitary adenomas' 2012). progression after surgery (Gejman et al., 2008).

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In general MKI67 is a useful marker in predicting 7. High MKI67 expression in colon cancer is tumor recurrence or invasiveness in accordance to associated with better relapse-free survival in other factors (de Aguiar et al., 2010). patients who underwent surgery and adjuvant Gastric cancer chemotherapy (Fluge et al., 2009). 8. MKI67 expression and the inflammation marker, Note C-reactive protein are correlated with poorer In gastric cancer the relation of MKI67 with the survival in patients undergoing surgery for characteristics of this malignancy are not well colorectal cancer (Canna et al., 2008). established. Expression of MKI67 and PCNA are correlated with high grade of gastric cancer Lung cancer (Czyzewska et al., 2004). The overexpression of Note TSPAN1 and MKI67 genes are negatively The MKI67 gene is not a validated prognostic correlated with carcinoma differentiation (Chen et marker in non-small cell lung cancer and more al., 2008). Additionally, MKI67 expression is studies with standardized methodology are required shown to be related to tumor size (Giaginis et al., to elucidate the role of this marker in this lung 2011). cancer subtype (Jakobsen and Sorensen, 2013). Gastrointestinal stromal tumor Lymphoma Note Note MKI67 expression is higher in the malignant In lymphoma malignancies, MKI67 is considered gastrointestinal stromal tumors than in benign ones, as a powerful tool in distinguishing benign from indicating a role in predicting malignant potential malignant cases (Bryant et al., 2006). Additionally, of GIST in accordance with other tumor markers MKI67 can be used as a prognostic factor of such as TP53 and KIT (c-kit) (Aoyagi et al., 2009). survival, with highly expressing tumors being Addditionally, MKI67 expression was correlated correlated with worse overall survival (Kim et al., with GIST recurrence and patient survival (Belev et 2007). However, in specific subtypes such as al., 2013; Liu et al., 2013a). diffuse large B-cell lymphoma low expression of Colorectal cancer MKI67 is an adverse prognostic factor (Hasselblom et al., 2008). Another use of MKI67 expression is Note the evaluation of prognosis after therapy in specific In colorectal cancer, the MKI67 gene is associated lymphoma subtypes (Determann et al., 2008). with several characteristics of this malignancy. Additionally, there is a strong correlation of gene Neuroendocrine tumors expression with the response of this cancer to Note potential therapeutic approaches. More specifically: In neuroendocrine tumors, MKI67 expression is the 1. MKI67 in addition to other tumor markers such sole strong independent risk factor for poor as CD34, KRT19 (CK19), KRT20 (CK20) are outcome and it is considered as predictor marker of highly expressed in colorectal cancer compared to disease progression and it is significantly associated normal tissues. Additionally, expression of CD34 with patients' overall survival (Panzuto et al., 2012). and MKI67 are significantly correlated with tumor The use of MKI67 expression is believed to be a stage, differentiation and low survival (Ma et al., better prognostic marker than mitotic count in these 2010). malignancies (Khan et al., 2013). This gene is 2. MKI67 and BMI1 overexpression in colorectal incorporated in the grading of neuroendocrine cancer is significantly related to tumorigenesis, neoplasms into two major categories: a) well metastasis and prognosis (Lin et al., 2008). differentiated neuroendocrine tumors (grade 1 and 3. MKI67 expression is a reliable marker for 2) and b) poorly differentiated endocrine predicting metastatic potential of rectal carcinoid carcinomas (grade 3) (Fung et al., 2013). In ileal (Hotta et al., 2006). well-differentiated neuroendocrine tumors, MKI67 4. The expression of MKI67, PDPN (podoplanin) predicts the progression-free survival (Dhall et al., and cytokeratin are correlated with increased 2012). However, a differentiation about the lympangiogenesis but not with poorer prognosis prognostic value of this gene is reported for the (Omachi et al., 2007). small cell lung patients where it has no prognostic 5. Decreased expression of MKI67, HIF1A and value (Erler et al., 2011). BCL2 markers were found after chemoradiotherapy of rectal cancer (Havelund et al., 2013). Thyroid cancer 6. Rectal tumors with lower MKI67 expression Note were more sensitive to neoadjuvant therapy and the In thyroid carcinomas, MKI67 and TP53 are gene expression was decreased after the therapy expressed with increasing frequency in progressed (Jiang et al., 2008). stages of this cancer (Saltman et al., 2006).

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MKI67 expression and mutational analysis of RET Schlüter C, Duchrow M, Wohlenberg C, Becker MH, Key gene could be used as prognostic factors of more G, Flad HD, Gerdes J. The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear aggressive medullary thyroid carcinoma (Mian et protein with numerous repeated elements, representing a al., 2011). MKI67 expression, LGALS3 (Galectin- new kind of cell cycle-maintaining proteins. J Cell Biol. 3) and PTTG1 (pituitary tumor transforming 1) 1993 Nov;123(3):513-22 genes could be used not only to distinguish benign Ross W, Hall PA. Ki67: from antibody to molecule to from malignant tumors but also to differentiate understanding? Clin Mol Pathol. 1995 Jun;48(3):M113-7 follicular carcinoma from papillary one (Cui et al., Duchrow M, Schlüter C, Wohlenberg C, Flad HD, Gerdes 2012). J. Molecular characterization of the gene locus of the human cell proliferation-associated nuclear protein defined Other cancers by monoclonal antibody Ki-67. Cell Prolif. 1996 Note Jan;29(1):1-12 Basal cell carcinoma, cutaneous pilar leiomyoma, Kill IR. Localisation of the Ki-67 antigen within the leiomyosarcoma, embryonal tumors, gallbladder nucleolus. Evidence for a fibrillarin-deficient region of the carcinoma, hypopharyngeal-squamous cell dense fibrillar component. J Cell Sci. 1996 Jun;109 ( Pt 6):1253-63 carcinoma, squamous cell carcinoma of larynx, malignant fibrous histiocytoma, parathyroid cancer, Starborg M, Gell K, Brundell E, Höög C. The murine Ki-67 cell proliferation antigen accumulates in the nucleolar and hepatocellular carcinoma, prostate cancer, penile heterochromatic regions of interphase cells and at the carcinoma, retinoblastoma and salivary gland periphery of the mitotic chromosomes in a process tumors are some of the malignancies in which essential for cell cycle progression. J Cell Sci. 1996 MKI67 expression is considered as proliferation, Jan;109 ( Pt 1):143-53 stage, grade or survival marker. However, more Bridger JM, Kill IR, Lichter P. Association of pKi-67 with sophisticated studies are needed in order to satellite DNA of the human genome in early G1 cells. elucidate the exact correlation of this gene with Chromosome Res. 1998 Jan;6(1):13-24 these tumors. Gassel AM, Backe J, Krebs S, Schön S, Caffier H, Müller- Hermelink HK. Endometrial carcinoma: Other non-malignant diseases immunohistochemically detected proliferation index is a Note prognosticator of long-term outcome. J Clin Pathol. 1998 Jan;51(1):25-9 MKI67 is evaluated in several other not malignant diseases, mostly as a proliferation marker, such as Durocher D, Henckel J, Fersht AR, Jackson SP. The FHA domain is a modular phosphopeptide recognition motif. hemimegalencephaly, eosinophilic esophagitis, Mol Cell. 1999 Sep;4(3):387-94 hepatitis, Reinke's edema, problems in placental development, psoriasis, Barret's esophagus, Plaat B, Kole A, Mastik M, Hoekstra H, Molenaar W, Vaalburg W. Protein synthesis rate measured with L-[1- endometriosis, multiple sclerosis, hemangioma, 11C]tyrosine positron emission tomography correlates with myocardial infarction, Alzheimer disease, mitotic activity and MIB-1 antibody-detected proliferation in Langerhans cell histiocytosis and endometrial human soft tissue sarcomas. Eur J Nucl Med. 1999 polyps. Apr;26(4):328-32 Salvesen HB, Iversen OE, Akslen LA. Prognostic References significance of angiogenesis and Ki-67, p53, and p21 expression: a population-based endometrial carcinoma Walker JE, Saraste M, Runswick MJ, Gay NJ. Distantly study. J Clin Oncol. 1999 May;17(5):1382-90 related sequences in the alpha- and beta-subunits of ATP Takagi M, Matsuoka Y, Kurihara T, Yoneda Y. Chmadrin: a synthase, myosin, kinases and other ATP-requiring novel Ki-67 antigen-related perichromosomal protein enzymes and a common nucleotide binding fold. EMBO J. possibly implicated in higher order chromatin structure. J 1982;1(8):945-51 Cell Sci. 1999 Aug;112 ( Pt 15):2463-72 Gerdes J, Schwab U, Lemke H, Stein H. Production of a Kreitz S, Fackelmayer FO, Gerdes J, Knippers R. The mouse monoclonal antibody reactive with a human nuclear proliferation-specific human Ki-67 protein is a constituent antigen associated with cell proliferation. Int J Cancer. of compact chromatin. Exp Cell Res. 2000 Nov 1983 Jan 15;31(1):13-20 25;261(1):284-92 Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Sueishi M, Takagi M, Yoneda Y. The forkhead-associated Stein H. Cell cycle analysis of a cell proliferation- domain of Ki-67 antigen interacts with the novel kinesin- associated human nuclear antigen defined by the like protein Hklp2. J Biol Chem. 2000 Sep monoclonal antibody Ki-67. J Immunol. 1984 15;275(37):28888-92 Oct;133(4):1710-5 Takagi M, Sueishi M, Saiwaki T, Kametaka A, Yoneda Y. A Verheijen R, Kuijpers HJ, van Driel R, Beck JL, van novel nucleolar protein, NIFK, interacts with the forkhead Dierendonck JH, Brakenhoff GJ, Ramaekers FC. Ki-67 associated domain of Ki-67 antigen in mitosis. J Biol detects a nuclear matrix-associated proliferation-related Chem. 2001 Jul 6;276(27):25386-91 antigen. II. Localization in mitotic cells and association with chromosomes. J Cell Sci. 1989 Apr;92 ( Pt 4):531-40 Brown DC, Gatter KC. Ki67 protein: the immaculate deception? Histopathology. 2002 Jan;40(1):2-11 Yasuda Y, Maul GG. A nucleolar auto-antigen is part of a major chromosomal surface component. Chromosoma. Brustmann H, Naudé S. Expression of topoisomerase 1990 Apr;99(2):152-60 IIalpha, Ki-67, proliferating cell nuclear antigen, p53, and

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 113 MKI67 (marker of proliferation Ki-67) Ntzeros K, et al.

argyrophilic nucleolar organizer regions in vulvar neoplasia]. Nan Fang Yi Ke Da Xue Xue Bao. 2007 squamous lesions. Gynecol Oncol. 2002 Aug;86(2):192-9 Apr;27(4):515-7 Kametaka A, Takagi M, Hayakawa T, Haraguchi T, Woenckhaus J, Steger K, Sturm K, Münstedt K, Franke Hiraoka Y, Yoneda Y. Interaction of the chromatin FE, Fenic I. Prognostic value of PIK3CA and compaction-inducing domain (LR domain) of Ki-67 antigen phosphorylated AKT expression in ovarian cancer. with HP1 proteins. Genes Cells. 2002 Dec;7(12):1231-42 Virchows Arch. 2007 Apr;450(4):387-95 Korkolopoulou P, Vassilopoulos I et al.. The combined Canna K, Hilmy M, McMillan DC, Smith GW, McKee RF, evaluation of p27Kip1 and Ki-67 expression provides McArdle CS, McNicol AM. The relationship between independent information on overall survival of ovarian tumour proliferative activity, the systemic inflammatory carcinoma patients. Gynecol Oncol. 2002 Jun;85(3):404- response and survival in patients undergoing curative 14 resection for colorectal cancer. Colorectal Dis. 2008 Sep;10(7):663-7 Scholzen T, Endl E, Wohlenberg C, van der Sar S, Cowell IG, Gerdes J, Singh PB. The Ki-67 protein interacts with Chen L, Li X, Wang GL, Wang Y, Zhu YY, Zhu J. members of the heterochromatin protein 1 (HP1) family: a Clinicopathological significance of overexpression of potential role in the regulation of higher-order chromatin TSPAN1, Ki67 and CD34 in gastric carcinoma. Tumori. structure. J Pathol. 2002 Feb;196(2):135-44 2008 Jul-Aug;94(4):531-8 Hammet A, Pike BL, McNees CJ, Conlan LA, Tenis N, Determann O, Hoster E, Ott G, Wolfram Bernd H, Heierhorst J. FHA domains as phospho-threonine binding Loddenkemper C, Leo Hansmann M, Barth TE, Unterhalt modules in cell signaling. IUBMB Life. 2003 Jan;55(1):23-7 M, Hiddemann W, Dreyling M, Klapper W. Ki-67 predicts outcome in advanced-stage mantle cell lymphoma patients Bubán T, Schmidt M, Broll R, Antal-Szalmás P, Duchrow treated with anti-CD20 immunochemotherapy: results from M. Detection of mutations in the cDNA of the proliferation randomized trials of the European MCL Network and the marker Ki-67 protein in four tumor cell lines. Cancer Genet German Low Grade Lymphoma Study Group. Blood. 2008 Cytogenet. 2004 Feb;149(1):81-4 Feb 15;111(4):2385-7 Czyzewska J, Guzi ńska-Ustymowicz K, Lebelt A, Zalewski Gejman R, Swearingen B, Hedley-Whyte ET. Role of Ki-67 B, Kemona A. Evaluation of proliferating markers Ki-67, proliferation index and p53 expression in predicting PCNA in gastric cancers. Rocz Akad Med Bialymst. progression of pituitary adenomas. Hum Pathol. 2008 2004;49 Suppl 1:64-6 May;39(5):758-66 Paek KI, Kim SH, Song SH, Choi SW, Koh HS, Youm JY, Hasselblom S, Ridell B, Sigurdardottir M, Hansson U, Kim Y. Clinical significance of Ki-67 labeling index in Nilsson-Ehle H, Andersson PO. Low rather than high Ki-67 pituitary macroadenoma. J Korean Med Sci. 2005 protein expression is an adverse prognostic factor in Jun;20(3):489-94 diffuse large B-cell lymphoma. Leuk Lymphoma. 2008 Bryant RJ, Banks PM, O'Malley DP. Ki67 staining pattern Aug;49(8):1501-9 as a diagnostic tool in the evaluation of lymphoproliferative Hoevenaars BM, van der Avoort IA, de Wilde PC, disorders. Histopathology. 2006 Apr;48(5):505-15 Massuger LF, Melchers WJ, de Hullu JA, Bulten J. A panel Hotta K, Shimoda T, Nakanishi Y, Saito D. Usefulness of of p16(INK4A), MIB1 and p53 proteins can distinguish Ki-67 for predicting the metastatic potential of rectal between the 2 pathways leading to vulvar squamous cell carcinoids. Pathol Int. 2006 Oct;56(10):591-6 carcinoma. Int J Cancer. 2008 Dec 15;123(12):2767-73 Saltman B, Singh B, Hedvat CV, Wreesmann VB, Jiang SM, Wang RB, Yu JM, Zhu KL, Mu DB, Xu ZF. Ghossein R. Patterns of expression of cell cycle/apoptosis [Correlation of VEGF and Ki67 expression with sensitivity genes along the spectrum of thyroid carcinoma to neoadjuvant chemoradiation in rectal adenocarcinoma]. progression. Surgery. 2006 Dec;140(6):899-905; Zhonghua Zhong Liu Za Zhi. 2008 Aug;30(8):602-5 discussion 905-6 Lin MX, Wen ZF, Feng ZY, He D. [Expression and Khouja MH, Baekelandt M, Nesland JM, Holm R. The significance of Bmi-1 and Ki67 in colorectal carcinoma clinical importance of Ki-67, p16, p14, and p57 expression tissues]. Ai Zheng. 2008 Dec;27(12):1321-6 in patients with advanced ovarian carcinoma. Int J Gynecol Looi ML, Dali AZ, Ali SA, Ngah WZ, Yusof YA. Expression Pathol. 2007 Oct;26(4):418-25 of p53, bcl-2 and Ki-67 in cervical intraepithelial neoplasia Kim SJ, Kim BS, Choi CW, Choi J, Kim I, Lee YH, Kim JS. and invasive squamous cell carcinoma of the uterine Ki-67 expression is predictive of prognosis in patients with cervix. Anal Quant Cytol Histol. 2008 Apr;30(2):63-70 stage I/II extranodal NK/T-cell lymphoma, nasal type. Ann Moorhead GB, Trinkle-Mulcahy L, Nimick M, De Wever V, Oncol. 2007 Aug;18(8):1382-7 Campbell DG, Gourlay R, Lam YW, Lamond AI. Miele A, Medina R, van Wijnen AJ, Stein GS, Stein JL. The Displacement affinity chromatography of protein interactome of the histone gene regulatory factor HiNF-P phosphatase one (PP1) complexes. BMC Biochem. 2008 suggests novel cell cycle related roles in transcriptional Nov 10;9:28 control and RNA processing. J Cell Biochem. 2007 Sep Terzi A, Saglam EA, Barak A, Soylemezoglu F. The 1;102(1):136-48 significance of immunohistochemical expression of Ki-67, Omachi T, Kawai Y, Mizuno R, Nomiyama T, Miyagawa S, p53, p21, and p16 in meningiomas tissue arrays. Pathol Ohhashi T, Nakayama J. Immunohistochemical Res Pract. 2008;204(5):305-14 demonstration of proliferating lymphatic vessels in Uzüm N, Atao ğlu GA. Histopathological parameters with colorectal carcinoma and its clinicopathological Ki-67 and bcl-2 in the prognosis of meningiomas according significance. Cancer Lett. 2007 Feb 8;246(1-2):167-72 to WHO 2000 classification. Tumori. 2008 May- Shi J, Zheng JS, Yin F, Hu WW, Huang XJ, Zhou XL. Jun;94(3):389-97 [Association of p16, p53, Ki-67 expressions with high-risk Ancu ţa E, Ancu ţa C, Cozma LG, Iordache C, Anghelache- human papilloma virus infection in cervical intraepithelial Lupa şcu I, Anton E, Carasevici E, Chirieac R. Tumor

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 114 MKI67 (marker of proliferation Ki-67) Ntzeros K, et al.

biomarkers in cervical cancer: focus on Ki-67 proliferation Gatta LB, Berenzi A, Balzarini P, Dessy E, Angiero F, factor and E-cadherin expression. Rom J Morphol Alessandri G, Gambino A, Grigolato P, Benetti A. Embryol. 2009;50(3):413-8 Diagnostic implications of L1, p16, and Ki-67 proteins and HPV DNA in low-grade cervical intraepithelial neoplasia. Aoyagi K, Kouhuji K, Yano S, Miyagi M, Imaizumi T, Int J Gynecol Pathol. 2011 Nov;30(6):597-604 Takeda J, Shirouzu K. Malignant potential of gastrointestinal stromal tumor of the stomach. Int Surg. Giaginis C, Giagini A, Tsourouflis G, Gatzidou E, Agapitos 2009 Jan-Feb;94(1):1-9 E, Kouraklis G, Theocharis S. MCM-2 and MCM-5 expression in gastric adenocarcinoma: clinical significance Conesa-Zamora P, Doménech-Peris A, Orantes-Casado and comparison with Ki-67 proliferative marker. Dig Dis FJ, Ortiz-Reina S, Sahuquillo-Frías L, Acosta-Ortega J, Sci. 2011 Mar;56(3):777-85 García-Solano J, Pérez-Guillermo M. Effect of human papillomavirus on cell cycle-related proteins p16, Ki-67, Huang L, Jiang T, Yuan F, Li GL, Xu LX, Cui Y. Cyclin D1, p53, and ProEx C in precursor lesions of [Correlation between loss of heterozygosity on cervical carcinoma: a tissue microarray study. Am J Clin chromosome 1p and 19q and expression of MGMT, p53 Pathol. 2009 Sep;132(3):378-90 and Ki-67 proteins in gliomas]. Zhonghua Zhong Liu Za Zhi. 2011 Oct;33(10):752-8 Fluge Ø, Gravdal K, Carlsen E, Vonen B, Kjellevold K, Refsum S, Lilleng R, Eide TJ, Halvorsen TB, Tveit KM, Keam B, Im SA, Lee KH, Han SW, Oh DY, Kim JH, Lee Otte AP, Akslen LA, Dahl O. Expression of EZH2 and Ki- SH, Han W, Kim DW, Kim TY, Park IA, Noh DY, Heo DS, 67 in colorectal cancer and associations with treatment Bang YJ. Ki-67 can be used for further classification of response and prognosis. Br J Cancer. 2009 Oct triple negative breast cancer into two subtypes with 20;101(8):1282-9 different response and prognosis. Breast Cancer Res. 2011 Mar 2;13(2):R22 Gincheva D, Tomov C, Gorchev G, Nikolova M. [The expression of p16(INK4a) and Ki-67 in atrophic- Mian C, Pennelli G, Barollo S, Cavedon E, Nacamulli D, hyperplastic and dysdysplastic lesions of the vulva]. Akush Vianello F, Negro I, Pozza G, Boschin IM, Pelizzo MR, Ginekol (Sofiia). 2009;48(2):16-20 Rugge M, Mantero F, Girelli ME, Opocher G. Combined RET and Ki-67 assessment in sporadic medullary thyroid Kosugi S, Hasebe M, Matsumura N, Takashima H, carcinoma: a useful tool for patient risk stratification. Eur J Miyamoto-Sato E, Tomita M, Yanagawa H. Six classes of Endocrinol. 2011 Jun;164(6):971-6 nuclear localization signals specific to different binding grooves of importin alpha. J Biol Chem. 2009 Jan Shen XX, Yu L, Bi R, Yang WT. [Clinicopathologic study 2;284(1):478-85 and immunohistochemistry comparison of Pax2, p53 and Ki-67 in low- and high-grade ovarian serous carcinomas]. Park S, Suh YL. Useful immunohistochemical markers for Zhonghua Bing Li Xue Za Zhi. 2011 Aug;40(8):511-6 distinguishing Paget cells from Toker cells. Pathology. 2009;41(7):640-4 Stoian SC, Simionescu C, M ărg ăritescu C, Stepan A, Nurciu M. Endometrial carcinomas: correlation between Vasilescu F, Ceau şu M, T ănase C, St ănculescu R, ER, PR, Ki67 status and histopathological prognostic Vl ădescu T, Ceau şu Z. P53, p63 and Ki-67 assessment in parameters. Rom J Morphol Embryol. 2011;52(2):631-6 HPV-induced cervical neoplasia. Rom J Morphol Embryol. 2009;50(3):357-61 Tanei T, Shimomura A, Shimazu K, Nakayama T, Kim SJ, Iwamoto T, Tamaki Y, Noguchi S. Prognostic significance de Aguiar PH, Aires R, Laws ER, Isolan GR, Logullo A, of Ki67 index after neoadjuvant chemotherapy in breast Patil C, Katznelson L. Labeling index in pituitary adenomas cancer. Eur J Surg Oncol. 2011 Feb;37(2):155-61 evaluated by means of MIB-1: is there a prognostic role? A critical review. Neurol Res. 2010 Dec;32(10):1060-71 Cui W, Lu X, Zheng S, Ma Y, Liu X, Zhang W. The use of a combination of Ki-67, Galectin-3, and PTTG can Ma YL, Peng JY, Zhang P, Liu WJ, Huang L, Qin HL. distinguish the benign and malignant thyroid tumor. Clin Immunohistochemical analysis revealed CD34 and Ki67 Lab. 2012;58(5-6):419-26 protein expression as significant prognostic factors in colorectal cancer. Med Oncol. 2010 Jun;27(2):304-9 Dhall D, Mertens R, Bresee C, Parakh R, Wang HL, Li M, Dhall G, Colquhoun SD, Ines D, Chung F, Yu R, Nissen Markova I, Duskova M, Lubusky M, Kudela M, Zapletalová NN, Wolin E. Ki-67 proliferative index predicts progression- J, Procházka M, Pilka R. Selected immunohistochemical free survival of patients with well-differentiated ileal prognostic factors in endometrial cancer. Int J Gynecol neuroendocrine tumors. Hum Pathol. 2012 Apr;43(4):489- Cancer. 2010 May;20(4):576-82 95 Wang CL, Mei JH, Wang SS, Xu S, Xu LL, Xiong YF. Ding XH, Hui YZ, Lu LJ, Yang ZC, Yao CJ, Sun LJ, Chen [Expression of HER2/neu in meningiomas: an ZH, Shi Z. [Vulvar intraepithelial neoplasia: a immunohistochemistry and fluorescence in situ clinicopathologic study of twenty cases]. Zhonghua Bing Li hybridization study]. Zhonghua Bing Li Xue Za Zhi. 2010a Xue Za Zhi. 2012 Jun;41(6):382-5 Mar;39(3):156-60 Horbinski C, Nikiforova MN, Hagenkord JM, Hamilton RL, Wang S, Ma XY, Xia Y, Zhang LH. [Expressions of Ki67, Pollack IF. Interplay among BRAF, p16, p53, and MIB1 in PCNA and mitotic index in ovarian epithelial tumors]. pediatric low-grade gliomas. Neuro Oncol. 2012 Sichuan Da Xue Xue Bao Yi Xue Ban. 2010b Jun;14(6):777-89 Jul;41(4):575-80 Mourtzikou A, Kosmas K, Marouga A, Stamouli M, Zambon AC. Use of the Ki67 promoter to label cell cycle Pouliakis A, Karakitsos P. The use of an entry in living cells. Cytometry A. 2010 Jun;77(6):564-70 immunocytochemical double-labeling staining can display Erler BS, Presby MM, Finch M, Hodges A, Horowitz K, the distribution of Bcl-2/Ki-67 cells in endometrial Topilow AA, Matulewicz T. CD117, Ki-67, and p53 predict adenocarcinomas as well as in normal endometrium. Clin survival in neuroendocrine carcinomas, but not within the Lab. 2012;58(1-2):133-44 subgroup of small cell lung carcinoma. Tumour Biol. 2011 Panzuto F, Campana D, Fazio N, Brizzi MP, Boninsegna Feb;32(1):107-11 L, Nori F, Di Meglio G, Capurso G, Scarpa A, Dogliotti L,

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 115 MKI67 (marker of proliferation Ki-67) Ntzeros K, et al.

De Braud F, Tomassetti P, Delle Fave G, Falconi M. Risk 14;108(9):1838-45 factors for disease progression in advanced jejunoileal neuroendocrine tumors. Neuroendocrinology. Konofaos P, Kontzoglou K, Parakeva P, Kittas C, Margari 2012;96(1):32-40 N, Giaxnaki E, Pouliakis M, Kouraklis G, Karakitsos P. The role of ThinPrep cytology in the investigation of ki-67 index, Pei DS, Qian GW, Tian H, Mou J, Li W, Zheng JN. p53 and HER-2 detection in fine-needle aspirates of breast Analysis of human Ki-67 gene promoter and identification tumors. J BUON. 2013 Apr-Jun;18(2):352-8 of the Sp1 binding sites for Ki-67 transcription. Tumour Biol. 2012 Feb;33(1):257-66 Konstantinos K, Marios S, Anna M, Nikolaos K, Efstratios P, Paulina A. Expression of Ki-67 as proliferation Preusser M, Hoeftberger R, Woehrer A, Gelpi E, biomarker in imprint smears of endometrial carcinoma. Kouwenhoven M, Kros JM, Sanson M, Idbaih A, Brandes Diagn Cytopathol. 2013 Mar;41(3):212-7 AA, Heinzl H, Gorlia T, Hainfellner JA, van den Bent M. Prognostic value of Ki67 index in anaplastic Kontzoglou K, Palla V, Karaolanis G, Karaiskos I, Alexiou oligodendroglial tumours--a translational study of the I, Pateras I, Konstantoudakis K, Stamatakos M. Correlation European Organization for Research and Treatment of between Ki67 and breast cancer prognosis. Oncology. Cancer Brain Tumor Group. Histopathology. 2012 2013;84(4):219-25 May;60(6):885-94 Koo YJ, Hahn HS, Lee IH, Lim KT, Lee KH, Kim HS, Kim Wentzensen N, Schwartz L, Zuna RE, Smith K, Mathews TJ, Chun YK, Kim HS, Hong SR. Dual immunostaining of C, Gold MA, Allen RA, Zhang R, Dunn ST, Walker JL, cervical cytology specimens with atypical squamous cells Schiffman M. Performance of p16/Ki-67 immunostaining to for p16/Ki-67 does not exclude the existence of a high- detect cervical cancer precursors in a colposcopy referral grade squamous intraepithelial lesion. Virchows Arch. population. Clin Cancer Res. 2012 Aug 1;18(15):4154-62 2013 Nov;463(5):689-96 Belev B, Br čić I, Prejac J, Golubi ć ZA, Vrbanec D, Božikov Kucuk U, Bayol U, Pala EE, Cumurcu S. Importance of J, Aleri ć I, Boban M, Razumovi ć JJ. Role of Ki-67 as a P53, Ki-67 expression in the differential diagnosis of prognostic factor in gastrointestinal stromal tumors. World benign/malignant phyllodes tumors of the breast. Indian J J Gastroenterol. 2013 Jan 28;19(4):523-7 Pathol Microbiol. 2013 Apr-Jun;56(2):129-34 Fung AD, Cohen C, Kavuri S, Lawson D, Gao X, Reid MD. Liu LC, Xu WT, Wu X, Zhao P, Lv YL, Chen L. Phosphohistone H3 and Ki-67 labeling indices in cytologic Overexpression of carbonic anhydrase II and Ki-67 specimens from well-differentiated neuroendocrine tumors proteins in prognosis of gastrointestinal stromal tumors. of the gastrointestinal tract and pancreas: a comparative World J Gastroenterol. 2013a Apr 28;19(16):2473-80 analysis using automated image cytometry. Acta Cytol. Liu Y, Tang K, Yan W, Wang Y, You G, Kang C, Jiang T, 2013;57(5):501-8 Zhang W. Identifying Ki-67 specific miRNA-mRNA Hassanein M, Huiart L, Bourdon V, Rabayrol L, Geneix J, interactions in malignant astrocytomas. Neurosci Lett. Nogues C, Peyrat JP, Gesta P, Meynard P, Dreyfus H, 2013b Jun 24;546:36-41 Petrot D, Lidereau R, Noguchi T, Eisinger F, Extra JM, Pavelin S, Becic K, Forempoher G, Mrklic I, Pogorelic Z, Viens P, Jacquemier J, Sobol H. Prediction of BRCA1 Titlic M, Andelinovic S. Expression of Ki-67 and p53 in germ-line mutation status in patients with breast cancer meningiomas. Neoplasma. 2013;60(5):480-5 using histoprognosis grade, MS110, Lys27H3, vimentin, and KI67. Pathobiology. 2013;80(5):219-27 von Minckwitz G, Schmitt WD, Loibl S, Müller BM, Blohmer JU, Sinn BV, Eidtmann H, Eiermann W, Gerber B, Tesch Havelund BM, Sørensen FB, Pløen J, Lindebjerg J, H, Hilfrich J, Huober J, Fehm T, Barinoff J, Rüdiger T, Spindler KL, Jakobsen A. Immunohistological expression Erbstoesser E, Fasching PA, Karn T, Müller V, Jackisch C, of HIF-1α, GLUT-1, Bcl-2 and Ki-67 in consecutive Denkert C. Ki67 measured after neoadjuvant biopsies during chemoradiotherapy in patients with rectal chemotherapy for primary breast cancer. Clin Cancer Res. cancer. APMIS. 2013 Feb;121(2):127-38 2013 Aug 15;19(16):4521-31 Heeran MC, Høgdall CK, Kjaer SK, Christensen L, Jensen Wang L, Feng C, Ding G, Zhou Z, Jiang H, Wu Z. A, Blaakaer J, Christensen IJ, Høgdall EV. Prognostic Relationship of TP53 and Ki67 expression in bladder value of tissue protein expression levels of MIB-1 (Ki-67) in cancer under WHO 2004 classification. J BUON. 2013 Danish ovarian cancer patients. From the 'MALOVA' Apr-Jun;18(2):420-4 ovarian cancer study. APMIS. 2013 Dec;121(12):1177-86 Kucukgoz Gulec U, Gumurdulu D, Guzel AB et al.. Inwald EC, Klinkhammer-Schalke M, Hofstädter F, Zeman Prognostic importance of survivin, Ki-67, and F, Koller M, Gerstenhauer M, Ortmann O. Ki-67 is a topoisomerase II α in ovarian carcinoma. Arch Gynecol prognostic parameter in breast cancer patients: results of a Obstet. 2014 Feb;289(2):393-8 large population-based cohort of a cancer registry. Breast Cancer Res Treat. 2013 Jun;139(2):539-52 Li M, Zhao L, Shen D, Li X, Wang J, Wei L. Clinical implications and prognostic value of single and combined Ishihara M, Mukai H, Nagai S, Onozawa M, Nihei K, biomarkers in endometrial carcinoma. Chin Med J (Engl). Shimada T, Wada N. Retrospective analysis of risk factors 2014;127(8):1459-63 for central nervous system metastases in operable breast cancer: effects of biologic subtype and Ki67 Pathmanathan N, Balleine RL, Jayasinghe UW et al.. The overexpression on survival. Oncology. 2013;84(3):135-40 prognostic value of Ki67 in systemically untreated patients with node-negative breast cancer. J Clin Pathol. 2014 Jakobsen JN, Sørensen JB. Clinical impact of ki-67 Mar;67(3):222-8 labeling index in non-small cell lung cancer. Lung Cancer. 2013 Jan;79(1):1-7 This article should be referenced as such: Khan MS, Luong TV, Watkins J, Toumpanakis C, Caplin Ntzeros K, Stanier P, Mazis D, Kritikos N, Rozis M, ME, Meyer T. A comparison of Ki-67 and mitotic count as Anesidis E, Antoniou C, Stamatakos M. MKI67 (marker of prognostic markers for metastatic pancreatic and midgut proliferation Ki-67). Atlas Genet Cytogenet Oncol neuroendocrine neoplasms. Br J Cancer. 2013 May Haematol. 2015; 19(2):105-116.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 116

Atlas of Genetics and Cytogenetics

in Oncology and Haematology

OPEN ACCESS JOURNAL INIST -CNRS

Gene Section Short Communication

OSGIN1 (oxidative stress induced growth inhibitor 1) Jing Hu, Yanming Wang University of California, San Diego, USA (JH), Pennsylvania State University, USA (YW)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/OSGIN1ID45760ch16q23.html DOI: 10.4267/2042/56296 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Transcription Review on OSGIN1, with data on DNA/RNA, on OSGIN1 transcript contains 6 exons. ORF is 1434 the protein encoded and where the gene is bp, which expands from exon 2 to exon 6. And implicated. exon 6 also contributes to 371 bp 3'UTR. Identity Protein Other names: BDGI, OKL38 Note Human OSGIN1/BDGI has a dominant isoform HGNC (Hugo): OSGIN1 containing 477 amino acids with calculated 52 kDa Location: 16q23.3 molecular weight (derived from transcript variants HuOKL38-1a/2a). DNA/RNA Besides, there are a 61 kDa longer isoform of OSGIN1 with 560 amino acids (transcribed from Note variant HuOKL38-2b), a 59 kDa longer isoform of Human OSGIN1 is located on chromosome 16 in OSGIN1 with 560 amino acids (transcribed from the region of q23.3. variant HuOKL38-2c), and a 34 kDa shorter Description isoform with 317 amino acids, which was the first Human OSGIN1 gene is 13111 bp in length, one to identify in 2001 and might be generated by composed of 6 exons and 5 introns, and located at internal transcription start codon within ORF or chromosome 16q23.3. cleaved from longer isoforms.

OSGIN1 genomic organization and transcript. The schematic representation of human OSGIN1 gene and its transcript. ATG: translation start codon; TAA: translation stop codon; UTR: untranslated region; ORF: open reading frame (according to Ref-seq).

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The predicted OSGIN1 structure contains a potential flavoprotein involved in K+ transport domain (TrkA) (201 aa), a potential NAD(P)-binding Rossmann-like domain (NAD_binding_8) (134 aa), a potential pyridine nucleotide-disulphide oxidoreductase domain (Pyr_redox_2) (201 aa) and a potential putative bacillithiol system oxidoreductase, YpdA family domain (Bthiol_YpdA) (295 aa).

Description OSGIN1 is also up-regulated by p53, and then triggers apoptosis by changing mitochondrial OSGIN1, described as a pregnancy-induced growth morphology, elevating ROS levels and inducing inhibitor, belongs to OKL38 protein family. cytochrome c release. Thus, OKL38 likely plays a Expression critical role in multiple tissues to guard against OSGIN1 gene was identified in rat mammary tumorigenesis. secretory epithelial cells, which was up-regulated Homology significantly in mammary gland during pregnancy and lactation. In various human normal tissues, An alignment of the amino acid sequences in OSGIN1 transcripts are observed with basal levels, ClustalW showed that OSGIN1 and OSGIN2, but increase remarkably in liver, followed by another member of OKL38 family, share 49% kidney, ovary, testis, and spleen especially. On the sequence identity, especially in C-terminal region contrary, OSGIN1 show low or undetectable of the two proteins. mRNA expression in liver, kidney, ovary tumor However, the biological function of OSGIN2 tissues compared to their paired normal remains unclear so far, which may be implicated in counterparts. Similarly, it is rarely expressed in Nijmegen breakage syndrome and gastric cancer. many cancer cell lines, including HepG2, SL, NIH, U2OS, MCF-7. Mutations Localisation Somatic Nucleus and mitochondria. Based on analysis of tumor and adjacent Function noncancerous tissues from total 400 patients of OSGIN1 was first discovered in 2001 as a hepatocellular carcinoma (HCC), a single- pregnancy-induced growth inhibitor. It is highly nucleotide variation from G to A at nt 1494 of expressed in ovary, kidney and liver. Stable OSGIN1 was identified, resulting in an amino acid expression of OSGIN1 is characterized by substitution R438H at codon region. relatively low proliferative rate and extensive Although OSGIN1 1494A variant appears in both differentiation. Conversely, loss of OKL38 activity tumor and noncancerous tissues, it shows higher leads to a disruption in the balance between cell occurrence in the tumor tissues than the common growth, cell proliferation and cell death, and is variant 1494G. associated with rapid tumor growth. OSGIN1 is And the allele-specific imbalance of OSGIN1 at inducible by distinct stress signals in multiple cell nt1494 is highly possible due to its localization at types. Oxidative stress induced by oxidized chromosome 16q23.3, which is prone to loss of phospholipids (OxPAPC and its component lipid heterozygosity in a variety of cancers. PEIPC) mediates expression of OSGIN1 regulated Functional studies revealed that OSGIN1 1494A via Nox/Nrf2 pathway. After DNA damage variant may have defects in translocation to treatment in cancer cells such as MCF-7 and U2OS, mitochondria and apoptosis.

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Implicated in Renal cell carcinoma (RCC) Hepatocellular carcinoma (HCC) Disease Renal cell carcinoma (RCC) is the primary type of Disease the adult kidney cancer. In contrast to HCC, RCC HCC is the most common type of liver cancer and incidence rates are higher in North one of the top human malignances worldwide. America/Western Europe region but lower in Generally, chronic liver injury by cirrhosis and Asia/Africa region. The causes of RCC are infection of hepatitis viruses are two major causes complicated, probably due to lifestyle-related of HCC. HCC also show higher incidence in and/or hereditary factors. population of Asia/Africa than North Prognosis America/Western Europe, suggesting a variety of Both OKL38 transcripts and protein levels are low underlying genetic and environmental factors. or undetected in majority of the kidney tumors Prognosis examined compared to patient's corresponding OSGIN1 expression analysis between tumor and normal adjacent tissues using cancer profiling paired noncancerous tissues from 89 HCC patients assay, western blot assay and immunohistological with clinicopathological data indicated the patients analysis. with less OSGIN1 transcripts and higher occupancy Oncogenesis of OSGIN1 1494A variant have more sever Forced overexpression of OSGIN1 in human symptoms and shorter survival time. Thus, kidney cancer cell A498 inhibits cell growth and quantitation of OSGIN1 expression and presence of stimulates cell death. OSGIN1 1494A variant may help diagnose HCC patients at early clinical trail and their responses Breast cancer after chemotherapy. Disease Oncogenesis Breast cancer is the most common cancer and the OSGIN1 expression is higher in immortal liver cell second top cause of cancer death among women. lines (LO2 and Miha) that that in HCC cell lines The American Cancer Society's estimates about (PLC8024 and Hep3B). Loss of wild-type OSGIN1 232670 new cases of invasive breast cancer will be in HCC tumors at higher stages and HCC cell lines diagnosed in women in the US for 2014. Risk may be due to its 5' untranslated region (5'UTR)- factors associated with breast cancer include age, regulated mRNA translation suppression. And geography, family history and so on. Especially, OSGIN1 1494A variant coded protein is less genetic risk factors with mutations in some capable to translocate from nucleus to mitochondria signature genes (e.g. BRCA1 and BRCA2) have and induce apoptosis compared to its wide-type been studied well for prognosis and treatment of protein, suggesting its impaired role as tumor breast cancer. Interestingly, some particular suppressor. reproductive factors like earlier age at first full-term

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 119 OSGIN1 (oxidative stress induced growth inhibitor 1) Hu J, Wang Y

pregnancy and higher number of pregnancies can inhibits growth and migration of breast cancer cells via reduce breast cancer significantly. induction of cell cycle arrest and apoptosis. J Biol Chem. 2005 Feb 11;280(6):4374-82 Oncogenesis Li R, Chen W, Yanes R, Lee S, Berliner JA. OKL38 is an Overexpression of OSGIN1 in MCF-7 human oxidative stress response gene stimulated by oxidized breast adenocarcinoma cells decreases their in vitro phospholipids. J Lipid Res. 2007 Mar;48(3):709-15 metastatic activity and in vivo tumor formation. Ong CK, Leong C, Tan PH, Van T, Huynh H. The role of 5' The mechanistic study found that high expression untranslated region in translational suppression of OKL38 of OSGIN1 may direct cell cycle arrested in S mRNA in hepatocellular carcinoma. Oncogene. 2007 Feb phase, and induce apoptosis further. 22;26(8):1155-65 These data suggest a potential correlation between Yao H, Li P, Venters BJ, Zheng S, Thompson PR, Pugh OSGIN1 and the reduction in breast cancer risk BF, Wang Y. Histone Arg modifications and p53 regulate observed in pregnancy-associated cases. the expression of OKL38, a mediator of apoptosis. J Biol Chem. 2008 Jul 18;283(29):20060-8 References Hu J, Yao H, Gan F, Tokarski A, Wang Y. Interaction of OKL38 and p53 in regulating mitochondrial structure and Huynh H, Ng CY, Ong CK, Lim KB, Chan TW. Cloning and function. PLoS One. 2012;7(8):e43362 characterization of a novel pregnancy-induced growth Liu M, Li Y, Chen L, Chan TH, Song Y, Fu L, Zeng TT, Dai inhibitor in mammary gland. Endocrinology. 2001 YD, Zhu YH, Li Y, Chen J, Yuan YF, Guan XY. Allele- Aug;142(8):3607-15 specific imbalance of oxidative stress-induced growth Ong CK, Ng CY, Leong C, Ng CP, Foo KT, Tan PH, Huynh inhibitor 1 associates with progression of hepatocellular H. Genomic structure of human OKL38 gene and its carcinoma. Gastroenterology. 2014 Apr;146(4):1084-96 differential expression in kidney carcinogenesis. J Biol Chem. 2004 Jan 2;279(1):743-54 This article should be referenced as such: Wang T, Xia D, Li N, Wang C, Chen T, Wan T, Chen G, Hu J, Wang Y. OSGIN1 (oxidative stress induced growth Cao X. Bone marrow stromal cell-derived growth inhibitor inhibitor 1). Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2):117-120.

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PTPRA (protein tyrosine phosphatase, receptor type, A) Jian Huang, Xueping Lai, Xinmin Zheng Department of Biochemistry and Molecular Cell Biology, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China (JH, XL), Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA (XZ)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/PTPRAID41923ch20p13.html DOI: 10.4267/2042/56297 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Transcription Three transcript variants encoding different Review on PTPRA, with data on DNA/RNA isoforms have been found for PTPRA gene. expression, on the protein encoded, and the Transcript variant 1 (NM_002836.3) represents the functional importance of the gene. longest transcript and encodes the longer isoform1 including 802 aa residues. Transcript variant 2 Identity (NM_080840.2) contains an alternate 5' UTR exon which is different from transcript variant 1, and Other names: HEPTP, HLPR, HPTPA, lacks exons one to five and coding exon ten, when HPTPalpha, LRP, PTPA, PTPRL2, R-PTP-alpha, compared to variant 1. Transcript variant 3 RPTPA (NM_080841.2) contains an additional exon within HGNC (Hugo): PTPRA the 5' UTR, when compared to variant 2. Transcript Location: 20p13 variant 2 and transcript variant 3 encode the short isoform 2. Isoform 2 lacks a 9 aa internal segment, DNA/RNA because exon ten missing, compared to Isoform 1. It is known that the expression of RPTPa mRNA is Description increased by 2 to 10-fold in 70% (10 of 14) of late- Human PTPRA is located at Chromosome 20: stage colon tumors compared to normal colonic 2844830-3019722 bp. mucosa (Tabiti et al., 1995). Another study Its Entrez gene ID is 5786 (NCBI) or 9664 demonstrated that RPTPa mRNA was increased in (HGNC). 29% (15 of 51) of primary breast carcinomas and PTPRA consists of 21 coding exons (protein correlated with its protein overexpression (Ardini et isoform 1) or 20 coding exons (protein isoform 2). al., 2000). Recently, PTPRA mRNA splice mutants 82 organisms have orthologs with human gene were described from Chinese colon, breast, and PTPRA. liver tumors (Huang et al., 2011). The PTPRA gene is conserved across species Pseudogene including chimpanzee, Rhesus monkey, dog, cow, One pseudogene (ID: PGOHUM00000236674) for mouse, rat, chicken, zebrafish, and C. elegans. PTPRA described on the website Pseudogenes.org.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 121 PTPRA (protein tyrosine phosphatase, receptor type, A) Huang J, et al.

Schematic depiction of human gene PTPRA (upper panel), derived alternative transcripts (middle panel) and corresponding protein isoforms (lower panel). Arrows in the upper panel indicate the three distinct transcriptional start sites within chromosome 20p13. 28 exon numbers according to transcript variant 1 (NM_002836.3) are indicated above the corresponding boxes. In the middle panel the build-up of the three different PTPRA transcripts, deduced based on cDNA deposits in public databases, is depicted. Transcript variant 1 (NM_002836.3) represents the longest transcript and encodes the longer isoform 1. Transcript variant 2 (NM_080840.2) contains an alternate 5' UTR exon which is different from transcript variant 1, and lacks coding exons one to five and exon ten, when compared to variant 1. Transcript variant 3 (NM_080841.2) contains an additional exon within the 5' UTR, and lacks an coding exon and exon one to five, when compared to variant 1. Transcript variant 2 and transcript variant 3 encode the short isoform 2. The N-terminal, the protein domain reflects the signal peptide (SP). The transmembrane spanning regions (TM) and protein tyrosine phosphatase catalytic domain (D1 and D2) are shown respectively. Isoform 2 lacks a 9 aa internal segment, because of exon ten missing, compared to Isoform 1.

nervous system of mouse, especially in the dorsal Protein root ganglia, cranial ganglia and adrenal gland (den Hertog et al., 1996). During chicken development, Description chicken RPTPa (ChPTPa) is expressed in pre- The protein encoded by PTPRA gene is a member migratory and migrating granule cells, and in of the protein tyrosine phosphatase (PTP) family. Bergmann glia and their radial processes as This PTP contains an extracellular domain, a single determined by in situ hybridization and transmembrane segment and two tandem immunostaining (Fang et al., 1996). Taken together, intracytoplasmic catalytic domains, and thus these studies demonstrate that RPTPa is highly represents a receptor-type PTP. Three alternatively expressed in the developing brain of various species spliced variants of this gene are well known to (Shock et al., 1995; Yang and Friesel, 1998; den encode two distinct isoforms differing only in their Hertog et al., 1999). extracellular region. The shorter form, expressed in most tissues, has 793 aa of which 123 are Localisation extracellular. The longer form, RPTPa802, has nine Membrane. A typical single-pass type I membrane extra amino acids located just before the protein. transmembrane region and is expressed only in a Function few tissues, especially in brain. It is noted that extensive N- and O-linked glycosylation of RPTPa Generally, RPTPa has been shown to gives rise to a mature 130 kD form of the protein dephosphorylate and activate Src family kinases (Daum et al., 1994). (SFKs), and is implicated in the regulation of integrin signaling, cell adhesion and proliferation. Expression Studies involving overexpression of RPTPa were PTPRA gene was originally isolated by PCR-based the first to demonstrate that PTP can PTP identification and cloning from several groups dephosphorylate tyrosine 527 of Src and activate c- (Sap et al., 1990; Kaplan et al., 1990; Matthews et Src in vivo and in vitro. This activation causes al., 1990). The protein, RPTPa, is a widely cellular transformation (Zheng et al., 1992). distributed transmembrane molecule that is Dephosphorylation of tyrosine 527 of c-Src in particularly highly expressed in the brain and RPTPa overexpressing P19 embryonal carcinoma kidney (Sap et al., 1990). RPTPa is highly cells activates c-Src and induces neuronal expressed in the developing central and peripheral differentiation (den Hertog et al., 1993). This

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 122 PTPRA (protein tyrosine phosphatase, receptor type, A) Huang J, et al.

observation has been independent validated in studied involving RPTPa knockout mice that Implicated in exhibit a dramatic decrease (50-70%) in c-Src Breast cancer activity in the brain (Ponniah et al., 1999; Su et al., Note 1999). In addition to Src, RPTPa regulates other Src It is clear that RPTPa functions as an activator of c- family kinases. Src family kinases, and thus was considered to be As an example, Fyn dephosphorylation and an oncogene. However, the first study on human activation is observed in cells co-expressing RPTPa breast tumors by Ardini et al. revealed an and Fyn (Bhandari et al., 1998). unexpected and interesting role of RPTPa. RPTPa Additionally, dephosphorylation of c-Src, Fyn and protein levels are found significantly overexpressed Yes, but not Lyn, was observed in A431 cells in 29% of 51 samples. High RPTPa protein levels expressing RPTPa (Harder et al., 1998), indicating correlated with low tumor grade and positive some degree of specificity of RPTPa. estrogen receptor status. It is noteworthy that there is no evidence of gross Overexpression of RPTPa in MCF-7 breast cancer physical abnormalities in the RPTPa-deficient mice, cells (ER+) resulted in growth inhibition while indicating that RPTPa is not essential for activating c-Src (Ardini et al., 2000). In a later embryonic development. study, Zheng reported that knockdown of RPTPa One explanation is that certain functions of RPTPa and c-Src using RNAi induced apoptosis in are compensated for by other PTPs in mice estrogen receptor-negative breast cancer cells, but deficient in RPTPa (Pallen, 2003). not in immortalized noncancerous breast cells and RPTPa is involved in promoting integrin signaling ER-positive breast cancer cells (including MCF-7). through activation of SFKs. It is noted by Zheng that correlation between ER An earlier study shows that c-Src activation by status and c-Src/RPTPa dependence in breast RPTPa can increase the association of c-Src with cancer may be important for planning therapeutic focal adhesion kinase (FAK), and enhance tyrosine strategy (Zheng et al., 2008). Recently, Wang J and phosphorylation of the Src/FAK substrate paxillin colleagues reported that EGF-induced RPTPa (Harder et al., 1998). Recently, Sun et al. described phosphorylation at Ser180 and Ser204 in BT-20 a novel molecular complex of RPTPa-BCAR3-Cas- and SKBR3 breast cancer cell lines results in Src that is important in integrin signaling (Sun et increased c-Src kinase activity, due to a decrease in al., 2012). This complex forms in response to RPTPa binding with Grb2 and an increase in RPTPaTyr789 phosphorylation and mediates Cas RPTPa binding with c-Src. These observations localization to focal adhesions and Cas downstream reveal novel aspects of integration of an signaling to promote cell migration (Sun et al., EGF/PKC/RPTPa/Src pathway in breast cancer cell 2012). lines (Wang et al., 2013). Currently, Meyer et al. More recently, Cheng et al. identified two roles of demonstrate that RPTPa functions as a positive Grb2 in integrin signaling: one as a regulator of mediator of tumor initiation and maintenance in paxillin stability and upstream promoter of both HER2/Neu-positive breast tumors (Meyer et FAKTyr397 phosphorylation that is required for al., 2014). Src-FAK complex activation and another as an Colorectal cancer essential coordinator of RPTPa and activation of Note the Src-FAK interaction thus enabling the The first report correlation with RPTPa and colonic phosphorylation of RPTPaTyr789 (Cheng et al., tumors was reported by Tabiti et al. in 1995. They 2014). quantified mRNA levels of RPTPa from 14 colon carcinomas and compared these levels to adjacent Mutations healthy colon mucosa. They observed a 2 to 10-fold increase in mRNA levels in advanced (Dukes' stage Note D) carcinoma. Another study from Zheng et al. Huang et al. sequence RPTPa cDNAs from five reported that RNAi knockdown of RPTPa reduced types of Chinese human tumors and paired normal c-Src kinase activity in several colon cancer cell samples. They observed three sequences encoding lines (HCT-15, HCT-116 and HT-29), which truncated proteins, designated RPTPa245, suppresses anchorage-independent growth and RPTPa445, or RPTPa652, lacking the D1 domain induces apoptosis (Zheng et al., 2008). Recently, or both the D1 and D2 domains. One mutant, tissue-arrays containing 50 colorectal cancer RPTPa245, widely expressed in colon, breast, and specimens and 10 normal colon samples were liver tumors from individuals of Chinese origin, can analysed by immunohistochemistry for RPTPa form an RPTPa-RPTPa245 heterodimer and expression. In normal tissue samples, RPTPa activate c-Src. (Huang et al., 2011). expression was restricted to smooth muscle cells.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 123 PTPRA (protein tyrosine phosphatase, receptor type, A) Huang J, et al.

None of the normal colonocyte expressed RPTPa in tyrosine phosphatase. Nature. 1992 Sep measurable quantities. However, over 70% of the 24;359(6393):336-9 colon cancer samples demonstrated expression of den Hertog J, Pals CE, Peppelenbosch MP, Tertoolen LG, RPTPa (Krndija et al., 2010). These data provide de Laat SW, Kruijer W. Receptor protein tyrosine phosphatase alpha activates pp60c-src and is involved in evidence for an oncogenic role of RPTPa in neuronal differentiation. EMBO J. 1993 Oct;12(10):3789- colorectal cancer. 98 Gastric cancer Daum G, Regenass S, Sap J, Schlessinger J, Fischer EH. Multiple forms of the human tyrosine phosphatase RPTP Note alpha. Isozymes and differences in glycosylation. J Biol To date, only one study has revealed an association Chem. 1994 Apr 8;269(14):10524-8 between RPTPa expression and gastric cancer. Shock LP, Bare DJ, Klinz SG, Maness PF. Protein tyrosine RPTPa expression is observed in 44% of gastric phosphatases expressed in developing brain and retinal samples and was the most widely expressed of the Müller glia. Brain Res Mol Brain Res. 1995 Jan;28(1):110- five PTPs. Several clinicopathological features 6 were significantly linked with the expression of Tabiti K, Smith DR, Goh HS, Pallen CJ. Increased mRNA RPTPa, including gross appearance, expression of the receptor-like protein tyrosine lymphovascular invasion, lymph node metastasis, phosphatase alpha in late stage colon carcinomas. Cancer liver metastasis and peritoneal dissemination. (Wu Lett. 1995 Jul 13;93(2):239-48 et al., 2006). den Hertog J, Overvoorde J, de Laat SW. Expression of receptor protein-tyrosine phosphatase alpha mRNA and Oral squamous cell carcinoma protein during mouse embryogenesis. Mech Dev. 1996 (OSCC) Aug;58(1-2):89-101 Note Fang KS, Martins-Green M, Williams LT, Hanafusa H. Characterization of chicken protein tyrosine phosphatase In an earlier study, Berndt et al. evaluated RPTPa alpha and its expression in the central nervous system. expression in 12 oral squamous cell carcinoma Brain Res Mol Brain Res. 1996 Apr;37(1-2):1-14 (OSCC) samples. Interestingly, not only the tumor Bhandari V, Lim KL, Pallen CJ. Physical and functional cells but also the stromal fibro/myofibroblasts as interactions between receptor-like protein-tyrosine well as inflammatory cells account for RPTPa phosphatase alpha and p59fyn. J Biol Chem. 1998 Apr expression in OSCC. In particular, immunostaining 10;273(15):8691-8 revealed a predominantly intracellular pattern of Harder KW, Moller NP, Peacock JW, Jirik FR. Protein- RPTPa expression, which may be due to an tyrosine phosphatase alpha regulates Src family kinases incompletely glycosylated form (Berndt et al., and alters cell-substratum adhesion. J Biol Chem. 1998 Nov 27;273(48):31890-900 1999) and/or to proteolytic cleavage of RPTPa in vivo (Gil-Henn et al., 2001). Yang CQ, Friesel R. Identification of a receptor-like protein tyrosine phosphatase expressed during Xenopus Diffuse large B-cell lymphoma development. Dev Dyn. 1998 Jul;212(3):403-12 Cytogenetics Berndt A, Luo X, Böhmer FD, Kosmehl H. Expression of A dup(20)(p13p13) was found in a case of diffuse the transmembrane protein tyrosine phosphatase RPTPalpha in human oral squamous cell carcinoma. large B-cell lymphoma (Morin et al., 2013). Histochem Cell Biol. 1999 May;111(5):399-403 Hybrid/Mutated gene den Hertog J, Blanchetot C, Buist A, Overvoorde J, van TMC2/PTPRA. der Sar A, Tertoolen LG. Receptor protein-tyrosine phosphatase signalling in development. Int J Dev Biol. References 1999;43(7):723-33 Ponniah S, Wang DZ, Lim KL, Pallen CJ. Targeted Kaplan R, Morse B, Huebner K, Croce C, Howk R, Ravera disruption of the tyrosine phosphatase PTPalpha leads to M, Ricca G, Jaye M, Schlessinger J. Cloning of three constitutive downregulation of the kinases Src and Fyn. human tyrosine phosphatases reveals a multigene family Curr Biol. 1999 May 20;9(10):535-8 of receptor-linked protein-tyrosine-phosphatases expressed in brain. Proc Natl Acad Sci U S A. 1990 Su J, Muranjan M, Sap J. Receptor protein tyrosine Sep;87(18):7000-4 phosphatase alpha activates Src-family kinases and controls integrin-mediated responses in fibroblasts. Curr Matthews RJ, Cahir ED, Thomas ML. Identification of an Biol. 1999 May 20;9(10):505-11 additional member of the protein-tyrosine-phosphatase family: evidence for alternative splicing in the tyrosine Ardini E, Agresti R, Tagliabue E, Greco M, Aiello P, Yang phosphatase domain. Proc Natl Acad Sci U S A. 1990 LT, Ménard S, Sap J. Expression of protein tyrosine Jun;87(12):4444-8 phosphatase alpha (RPTPalpha) in human breast cancer correlates with low tumor grade, and inhibits tumor cell Sap J, D'Eustachio P, Givol D, Schlessinger J. Cloning growth in vitro and in vivo. Oncogene. 2000 Oct and expression of a widely expressed receptor tyrosine 12;19(43):4979-87 phosphatase. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6112-6 Gil-Henn H, Volohonsky G, Elson A. Regulation of protein- tyrosine phosphatases alpha and epsilon by calpain- Zheng XM, Wang Y, Pallen CJ. Cell transformation and mediated proteolytic cleavage. J Biol Chem. 2001 Aug activation of pp60c-src by overexpression of a protein 24;276(34):31772-9

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Pallen CJ. Protein tyrosine phosphatase alpha (PTPalpha): Huff RD, Scott DW, Ding J, Roth A, Chiu R, Corbett RD, a Src family kinase activator and mediator of multiple Chan FC, Mendez-Lago M, Trinh DL, Bolger-Munro M, biological effects. Curr Top Med Chem. 2003;3(7):821-35 Taylor G, Hadj Khodabakhshi A, Ben-Neriah S, Pon J, Meissner B, Woolcock B, Farnoud N, Rogic S, Lim EL, Wu CW, Kao HL, Li AF, Chi CW, Lin WC. Protein tyrosine- Johnson NA, Shah S, Jones S, Steidl C, Holt R, Birol I, phosphatase expression profiling in gastric cancer tissues. Moore R, Connors JM, Gascoyne RD, Marra MA. Cancer Lett. 2006 Oct 8;242(1):95-103 Mutational and structural analysis of diffuse large B-cell Zheng X, Resnick RJ, Shalloway D. Apoptosis of estrogen- lymphoma using whole-genome sequencing. Blood. 2013 receptor negative breast cancer and colon cancer cell lines Aug 15;122(7):1256-65 by PTP alpha and src RNAi. Int J Cancer. 2008 May Wang J, Yu L, Zheng X. PTP α-mediated Src activation by 1;122(9):1999-2007 EGF in human breast cancer cells. Acta Biochim Biophys Krndija D, Schmid H, Eismann JL, Lother U, Adler G, Sin (Shanghai). 2013 Apr;45(4):320-9 Oswald F, Seufferlein T, von Wichert G. Substrate stiffness Cheng SY, Sun G, Schlaepfer DD, Pallen CJ. Grb2 and the receptor-type tyrosine-protein phosphatase alpha promotes integrin-induced focal adhesion kinase (FAK) regulate spreading of colon cancer cells through autophosphorylation and directs the phosphorylation of cytoskeletal contractility. Oncogene. 2010 May protein tyrosine phosphatase α by the Src-FAK kinase 6;29(18):2724-38 complex. Mol Cell Biol. 2014 Feb;34(3):348-61 Huang J, Yao L, Xu R, Wu H, Wang M, White BS, Meyer DS, Aceto N, Sausgruber N, Brinkhaus H, Müller U, Shalloway D, Zheng X. Activation of Src and Pallen CJ, Bentires-Alj M. Tyrosine phosphatase PTPα transformation by an RPTP α splice mutant found in human contributes to HER2-evoked breast tumor initiation and tumours. EMBO J. 2011 Jul 1;30(15):3200-11 maintenance. Oncogene. 2014 Jan 16;33(3):398-402 Sun G, Cheng SY, Chen M, Lim CJ, Pallen CJ. Protein tyrosine phosphatase α phosphotyrosyl-789 binds BCAR3 This article should be referenced as such: to position Cas for activation at integrin-mediated focal Huang J, Lai X, Zheng X. PTPRA (protein tyrosine adhesions. Mol Cell Biol. 2012 Sep;32(18):3776-89 phosphatase, receptor type, A). Atlas Genet Cytogenet Morin RD, Mungall K, Pleasance E, Mungall AJ, Goya R, Oncol Haematol. 2015; 19(2):121-125.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 125

Atlas of Genetics and Cytogenetics

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Gene Section Review

STMN1 (stathmin 1) João Agostinho Machado-Neto, Fabiola Traina Hematology and Hemotherapy Center-University of Campinas/Hemocentro-Unicamp, Instituto Nacional de Ciencia e Tecnologia do Sangue, Campinas, Sao Paulo, Brazil (JAMN), Department of Internal Medicine, University of Sao Paulo at Ribeirao Preto Medical School, Ribeirao Preto, Sao Paulo, Brazil (FT)

Published in Atlas Database: May 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/STMN1ID42443ch1p36.html DOI: 10.4267/2042/56298 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

The STMN1 gene encodes 2 isoforms, A and B. Abstract Isoform A contains 3 transcript variants that differ Stathmin 1 (STMN1) is a microtubule destabilizer in the 5' UTR and have an alternate terminal exon, protein with an important role in cell cycle compared to isoform B, resulting in a shorter and progression, cell proliferation, migration and distinct C-terminus. The isoform B represents the survival. The present review on STMN1 contains longest transcript variant. data on DNA/RNA, on the protein encoded and where the gene is implicated. Protein Identity Description Stathmin 1 belongs to the Stathmin protein family, Other names: C1orf215, LAP18, Lag, OP18, which is characterized by the presence of a PP17, PP19, PR22, SMN Stathmin-like domain (also known as tubulin- HGNC (Hugo): STMN1 binding domain) that participates in Location: 1p36.11 interactions/sequestering of alpha/beta-tubulin heterodimers (Figure 1). DNA/RNA Expression Note Ubiquitous. Stathmin 1 is highly expressed during The entire STMN1 gene is about 22.8 kb and embryonic development. In adult cells, it is contains 5 exons (start: 26210672 bp and end: expressed during cell proliferation, and in nervous 26233482; orientation: minus strand). tissue and testis (revised in Curmi et al., 1999).

Figure 1. Representation of primary structure of Stathmin 1 protein. The catastrophe promotion region (aa 1 - 99) and the four serine phosphorylation sites (S16, S25, S38 and S63) at the N-terminal, and the tubulin binding domain (aa 25 - 149) at the C-terminal are illustrated in the figure. Reproduced with permission of the editor-in-chief of BMB reports from Machado-Neto et al., 2014b.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 126 STMN1 (stathmin 1) Machado-Neto JA, Traina F

Figure 2. Intracellular localization of Stathmin 1 protein in HeLa cells. Confocal analysis of HeLa cells displaying Stathmin 1 (green), Actin (red) and DAPI (blue) staining; MERGE shows the overlapped images. Scale bar: 10 µm. Note the predominant cytoplasmic localization of Stathmin 1. Anti-Stathmin 1 (OP18; sc-55531) was from Santa Cruz Biotechnology, (Santa Cruz, CA, USA), Phalloidin (A12379) and DAPI (P-36931) were from Invitrogen (Carlsbad, CA, USA). Personal data.

Localisation (PAK1) and Ca 2+ /calmodulin-dependent protein kinases (CamKs). Serine 25 and/or 38 may be Stathmin 1 is predominantly found in the cytoplasm phosphorylated by cyclin-dependent kinases (Figure 2). (CDKs), mitogen-activated protein kinases Function (MAPKs) and phosphoinositide 3-kinase (PI3K) Stathmin 1 is a phosphoprotein that participates in (Belletti and Baldassarre, 2011). microtubule catastrophe and/or in the sequestering Phosphatase proteins that are able to of alpha/beta-tubulin heterodimers, regulates dephosphorylate Stathmin 1 includes: protein microtubule dynamics, cell cycle progression, phosphatase 1 (PP1), protein phosphatase 2A proliferation, motility and survival (Curmi et al., (PP2A) and protein phosphatase 2B (PP2B) (Guy et 1999; Belletti and Baldassarre, 2011). The main al., 1992; Tournebize et al., 1997; Mistry et al., mechanism of regulation of Stathmin 1 activity on 1998) (Figure 3). based in its phophorylated and unphosphorylated Homology status at serine sites (residues 16, 25, 38 and 68). Stathmin 1 phosphorylation at serine 16 and/or 63 Stathmin 1 shares high homology with the other reduces the affinity between Stathmin 1 and members of the Stathmin protein family including alpha/beta-tubulin heterodimers. The proteins that Stathmin-like 2 (also named SCG10), Stathmin-like are able to phosphorylate Stathmin 1 at serine 16 3 (also named SCLIP) and Stathmin-like 4 (also and/or 63 are: aurora kinase B, protein kinase A named RB3). Stathmin 1 also shares high homology (PKA), P21 protein (Cdc42/Rac)-activated kinase among different species (Table 1).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 127 STMN1 (stathmin 1) Machado-Neto JA, Traina F

Figure 3. Stathmin 1 signaling. Stathmin 1 may be phosphorylated on serine sites by cyclin-dependent kinases (CDKs), mitogen-activated protein kinases (MAPKs), phosphoinositide 3-kinase (PI3K), aurora kinase B (AURKB), protein kinase A (PKA), and Ca 2+ /calmodulin-dependent protein kinases (CamKs), leading to microtubule stability. On the other hand, Stathmin 1 may be dephosphorylated by protein phosphatase 1 (PP1), protein phosphatase 2A (PP2A) and protein phosphatase 2B (PP2B), resulting in microtubule instability. Abbreviations: TKR: tyrosine kinase receptor; P: phosphorylation; Ac: acetylation. This figure was performed using Servier Medical Art tools (http://www.servier.com/Powerpoint-image-bank).

models, several groups have demonstrated that Mutations Stathmin 1 inhibition may partially reverse the Recurrent mutations in the STMN1 gene are rare, malignant phenotype. 10 missense, 8 nonsense, 1 nonstop extension and 3 Hematopoietic neoplasms coding silent mutations are reported at COSMIC (catalogue of somatic mutations in cancer; Note COSMIC). In human esophageal adenocarcinoma, Stathmin 1 was initially identified by proteomics STMN1 Q18E mutation was identified and analysis of HL60 leukemia cells (Feuerstein and presented transformation potential in vitro and in Cooper, 1983). In primary acute leukemia and vivo in NIH3T3 cells (Misek et al., 2002). lymphoma samples, and leukemia cell lines, several Studies performed in leukemia cells indicate that independent groups have showed that Stathmin 1 is the Q18E mutation results in Stathmin 1 highly expressed (Hanash et al., 1988; Melhem et hyperactivity and contributes to chromosomal al., 1991; Melhem et al., 1997; Roos et al., 1993; instability (Holmfeldt et al., 2006). Machado-Neto et al., 2014a; Brattsand et al., 1993); however its impact on survival outcome remains Implicated in unknown. A recent study of immunophenotypic and molecular features of a large series of follicular Various cancers lymphomas indicated that Stathmin 1 represents an Note useful novel diagnostic marker for these diseases The role and clinical impact of Stathmin 1 has been (Marafioti et al., 2013). Importantly, Stathmin 1 extensively addressed in many types of human silencing resulted in marked inhibition of cancers. In general, the increased expression of tumorigenicity, proliferation and clonogenicity in Stathmin 1 is found in many cancers and confers a leukemia cell lines (K562, U937 and Namalwa poor prognosis (revised in Belletti and Baldassarre, cells) (Machado-Neto et al., 2014a; Jeha et al., 2011 and Rana et al., 2008). Using cancer cell line 1996; Iancu et al., 2001).

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Table 1. Comparative identity of human STMN1 with other species. Source: HomoloGene.

In a study that focused on the identification of tumors (Li et al., 2011). Using breast cancer cell biomarkers of human aging and aging-related lines and gene therapy tools, Stathmin 1 inhibition, diseases, Stathmin 1 was found upregulated in by adenovirus-mediated gene transfer of anti- plasma samples from myelodysplastic patients Stathmin 1 ribozyme, resulted in a dose-dependent (Jiang et al., 2008). Recently, increased levels of inhibition of proliferation, apoptosis induction and Stathmin 1 were reported in bone marrow and had an additive effect together low concentration of CD34 + cells from high-risk myelodysplastic taxol treatment in vitro and in vivo (Miceli et al., syndromes patients and during disease progression 2013). (Machado-Neto et al., 2014a). Notably, higher Ovarian cancer Stathmin 1 expression was associated with high- risk disease and higher bone marrow blasts Note percentage (Machado-Neto et al., 2014a), but did Stathmin 1 overexpression has been described in not impacted survival. ovarian cancer patients (Alaiya et al., 1997; Price et Stathmin 1 was also identified as one of the 15 most al., 2000). Wei and colleagues (Wei et al., 2008) relevant genes for determining the outcome in observed that Stathmin 1 was expressed in all myeloma multiple patients by microarray approach ovarian cancer samples analyzed and higher levels (Decaux et al., 2008). were observed in the metastatic tumors and negatively impacted survival by univariate analysis. Breast cancer In agreement, Stathmin 1 overexpression was found Note in primary high-grade serous ovarian carcinomas Using Western blot analysis, Brattsand (Brattsand, and ovarian cancer cell lines (Karst et al., 2011). 2000) reported that Stathmin 1 expression High levels of Stathmin 1 predicted an unfavorable positively correlated with proliferation status and prognosis in ovarian cancer patients under aggressiveness in a panel of 151 primary breast paclitaxel and/or platinum therapy (Su et al., 2009; carcinoma samples. Similar results were found by Aoki et al., 2009) also by univariate analysis. In Curmi and colleagues (Curmi et al., 2000), who p53 mutated ovarian cancer cell lines, Stathmin 1 also reported that Stathmin 1 overexpression silencing caused cell cycle arrest and apoptosis in correlated with loss of steroid receptors, vivo and in vivo (Sonego et al., 2013). histopathological grade III and mitotic index. Head and neck cancer Importantly, Stathmin 1 expression was indicated as a potential tool for predicting the outcome of Note breast cancer patients with lymph node metastasis Using proteomics approach, Stathmin 1 was found and its expression was increased in the group with to be differently expressed in oral squamous-cell poor prognosis (Oishi et al., 2007). By multivariate carcinoma and laryngeal squamous-cell carcinoma analysis, high Stathmin 1 expression predicted (Koike et al., 2005; Sewell et al., 2007). Stathmin 1 reduced disease-free survival (Saal et al., 2007; expression was also found to be significantly Golouh et al., 2008; Baquero et al., 2012) and increased in oral squamous-cell carcinoma cell lines overall survival (Baquero et al., 2012). Decreased and primary tumors and its high expression Stathmin 1 phosphorylation at serine 16 (an correlated with advanced stages of the disease and inhibitory site) correlated with the more metastatic poor disease-free survival by univariate analysis phenotype in breast cancers cell lines and primary (Kouzu et al., 2006). In head-neck squamous-cell

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carcinoma, Stathmin 1 was expressed at low levels aggressive phenotype and worse survival. Of note, (76% cases) and did not impact recurrence Wik and colleagues (Wik et al., 2013) reported that (Canzonieri et al., 2012). Cheng and colleagues the levels of Stathmin 1 phosphorylation at serine (Cheng et al., 2008) identified an upregulation of 38 site were associated with poor prognosis, tumor Stathmin 1 in primary nasopharyngeal carcinoma cell proliferation, increased PIK3CA copy number and its expression was associated with recurrence and PI3K activation by multivariate analysis. and advanced stages of the disease. In agreement, Werner and colleagues (Werner et al., 2014), using Hsu and colleagues (Hsu et al., 2014) reported that endometrial cancer cell lines, showed that Stathmin Stathmin 1 was overexpressed in approximately 1 silencing potentiated the response to paclitaxel 50% of the nasopharyngeal carcinoma samples and treatment. This finding was confirmed in vivo: was associated with advanced age, high-grade endometrial cancer patients with high Stathmin 1 tumors and was an independent predictor of worse expression had a poor response to paclitaxel therapy disease-free survival. Notably, knockdown of (Werner et al., 2014). Stathmin 1 suppressed cell cycle progression, Bladder cancer and urothelial proliferation, migration, invasion, xenograft tumor growth, induced apoptosis and potentiated carcinoma paclitaxel response in nasopharyngeal carcinoma Note cell lines (CNE1-LMP1 and HNE2 cells) (Wu et Using quantitative PCR targeting 110 relevant al., 2014). cancer genes, Dubosq and colleagues (Dubosq et Hepatocarcinoma al., 2012) detected Stathmin 1 as highly expressed in early recurrence, compared to late or null Note recurrence cancer in a cohort of 47 bladder cancer Yuan and colleagues (Yuan et al., 2006) reported a patients, suggesting a role for this protein in the high Stathmin 1 expression in 56% of 156 time of recurrence. Bhagirath and colleagues hepatocarcinoma patients and high Stathmin 1 (Bhagirath et al., 2012) reported elevated STMN1 expression was associated with increased tumor mRNA levels in muscle invasive tumors. In size, tumor grade, metastasis, p53 mutation status agreement, patients with high Stathmin 1 and negatively impacted survival in univariate expression under taxane therapy had decreased analysis. In agreement, elevated Stathmin 1 levels recurrence-free survival by univariate analysis were also reported by Singer and colleagues (Singer (Wosnitzer et al., 2011). In a cohort of 58 urothelial et al., 2007) and were associated with the presence carcinoma patients, multivariate analysis revealed of undifferentiated tumors. Other studies observed that Stathmin 1 positivity was associated with high an increased Stathmin 1 expression in tumor tissue grade tumors, recurrence and negatively impacted compared to matched normal tissue, and a positive survival (Lin et al., 2009). association between Stathmin 1 overexpression and recurrence or poor prognosis in univariate analysis Colorectal cancer (Hsieh et al., 2010; Chen et al., 2013b). In Note hepatocarcinoma cell lines, Stathmin 1 silencing In a cohort of 149 patients with colorectal cancer, reduced cell proliferation, viability, migration and high Stathmin 1 levels were an independent augmented the response to paclitaxel, vinblastine predictor of worse overall survival (Zheng et al., and cisplatin treatment (Singer et al., 2007; Hsieh et 2010). In addition, Stathmin 1 expression was al., 2010; Gan et al., 2010). associated with tumor differentiation, invasion and stage of the disease (Zheng et al., 2010). In contrast, Endometrial cancer Ogino and colleagues (Ogino et al., 2009) showed Note that, by multivariate analysis, Stathmin 1 positivity In a multicenter study including 1076 endometrial had a protective effect on survival in a cohort of patients, Stathmin 1 overexpression was detected in 546 colorectal patients (stratified in obese and non- 37% of cases and correlated with high grade disease obese individuals). Interestingly, obesity had a and aneuploidy, and was an independent predictor negative impact on survival in Stathmin 1 positive of metastasis and worse disease specific survival patients, but not in Stathmin 1-negative patients (Trovik et al., 2011). In another study from the (Ogino et al., 2009). A recent study conduced by same group (Trovik et al., 2010), high Stathmin 1 Tan and colleagues (Tan et al., 2012) corroborated expression was associated with a higher probability the findings from Zheng and colleagues (Zheng et of endometrial cancer recurrence and PI3K al., 2010), indicating that high Stathmin 1 levels activation. Additionally, a study conducted by negatively impacted survival in a cohort of 324 Salvesen and colleagues (Salvesen et al., 2009), colorectal cancer patients in univariate analysis. investigating the impact of PI3K activation in Tan and colleagues (Tan et al., 2012) also endometrial cancer, identified that high Stathmin 1 demonstrated functional evidences that Stathmin 1 expression was an independent predictor of is a positive regulator of cell proliferation,

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clonogenicity, migration and invasion in colorectal Cervical cancer cancer cell lines. Note Gastric cancer A higher Stathmin 1 expression was found in Note primary cells and cell lines from cervical carcinoma Jeon and colleagues (Jeon et al., 2010) reported that compared to normal cervical epithelial cells and a high expression of Stathmin 1 was an independent also in tumor cells compared to matched adjacent non-carcinoma tissue (Xi et al., 2009). Increased predictor of shorter recurrence-free survival, and Stathmin 1 expression correlated with a worse associated with lymph node metastasis and high clinical stage and metastasis (Xi et al., 2009). grade stages in a cohort of 226 gastric cancer Another study found Stathmin 1 overexpression in patients. The authors, using two different gastric cancer cell lines (SNU638 and SNU16 cells), all cervical and rare expression of Stathmin 1 in demonstrated that Stathmin 1 silencing decreased benign samples; the authors suggest that the analysis of Stathmin 1 may be useful diagnostically cell proliferation, migration, invasion and xenograft in the identification of cervical cancer (Howitt et tumor growth (Jeon et al., 2010). Kang and al., 2013). colleagues (Kang et al., 2012), and Ke and colleagues (Ke et al., 2013), identified high Glioma Stathmin 1 expression in cell lines and primary Note cells from gastric cancer and predicted poor In a cohort of 24 glioma patients, increased prognosis by univariate analysis. Interestingly, Stathmin 1 levels were associated with decreased Kang and colleagues (Kang et al., 2012) also recurrence-free survival in univariate analysis (Ngo reported that Stathmin 1 silencing reduced the et al., 2007). Similar results were observed in a malignant phenotype in vitro and in vivo, and xenograft glioma nitrosourea-treated model (Ngo et suggested that miR-223 is involved in the al., 2007). Dong and colleagues (Dong et al., 2012) regulation of Stathmin 1 expression in gastric observed that Stathmin 1 expression was aberrantly cancer cell lines (AGS and MKN7 cells). Another expressed in vascular endothelial cells from glioma, study, using lentivirus mediated RNAi delivery, especially in high grade cases and the Stathmin 1 also demonstrated that Stathmin 1 silencing reduced silencing reduced cell proliferation and invasion, cell proliferation, migration and xenograft tumor and induced apoptosis in glioma-derived growth in MKN-45 gastric cancer cells (Akhtar et microvascular endothelial cells. al., 2013). Lung cancer Prostate cancer Note Note Chen and colleagues (Chen et al., 2003) reported a Using high-throughput immunoblotting, elevated high expression of Stathmin 1 in a cohort of 93 lung Stathmin 1 expression was found in metastatic adenocarcinoma patients and this expression was prostate cancer protein extracts (Varambally et al., associated with poorly differentiated tumors. 2005). Another study reported that Stathmin 1 Stathmin 1 overexpression was also found in expression was higher in advanced prostate tumors primary non-small cell lung tumors matched with (Ghosh et al., 2007). Stathmin 1 silencing resulted normal tissues (Singer et al., 2009). Rosell and in cell cycle arrest, reduced clonogenicity and colleagues (Rosell et al., 2003) observed that high increased apoptosis in prostate cancer cell line Stathmin 1 levels negatively affected the time to (LNCaP cells) (Mistry et al., 2005). In contrast, progression in non-small-cell lung cancer patients, Stathmin 1 inhibition augmented the epithelial-to- by univariate analysis. Of note, Stathmin 1 mesenchymal transition and metastasis potential in inhibition decreased proliferation, migration and another prostate cancer cell line (DU145 cells) invasion in non-small cell lung cancer cell lines (Williams et al., 2012). (Calu-1 and Calu-6 cells) (Singer et al., 2009). Pheochromocytomas Medulloblastoma Note Note In a study conducted by Sadow and colleagues Using a microarray approach, Stathmin 1 was (Sadow et al., 2008), among the endocrine tumors, identified as differentially expressed in primary high levels of Stathmin 1 were observed in medulloblastoma samples and it was associated malignant pheochromocytomas. These results were with unfavorable overall survival (Neben et al., confirmed by two other groups, who reported an 2004). Accordingly, Kuo and colleagues (Kuo et overexpression of Stathmin 1 in al., 2009) reported that Stathmin 1 correlated with malignant/metastatic pheochromocytomas tumor dissemination and predicted decreased compared to benign tumors or normal tissues survival in medulloblastoma patients, by univariate (Björklund et al., 2010; Lin et al., 2011). analysis in both studies.

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Pancreatic cancer highly expressed in primary primitive neuroectodermal tumors compared to Note ependymomas samples (de Bont et al., 2007). Lu and colleagues (Lu et al., 2014) reported that Stathmin 1 was overexpressed in pancreatic cancer Renal cancer samples and that high Stathmin 1 levels were Note correlated with vascular emboli, tumor size, and In Wilms tumors, Stathmin 1 was highly expressed negatively impacted overall survival in univariate in high grade compared to low grade tumors analysis. In addition, Stathmin 1 silencing in (Takahashi et al., 2002). pancreatic cancer cells resulted in reduced cell proliferation, clonogenicity and cell cycle arrest (Lu Sarcoma et al., 2014; Jiang et al., 2009). Note Thyroid cancer Belletti and colleagues (Belletti et al., 2008) Note reported that Stathmin 1 was increased in recurrent Using cDNA microarray approach, Onda and and metastatic sarcoma samples. colleagues (Onda et al., 2004) reported that In addition, overexpression of the wild type Stathmin 1 was overexpressed in all anaplastic Stathmin 1 or mutated Stathmin 1 (Q18E, gain-of- thyroid cancer cell lines analyzed and this was function mutation) potentiated the malignant confirmed by immunohistochemical analyses in phenotype in the sarcoma cell line HT1080 (Belletti primary samples. Another study also observed that et al., 2008). Stathmin 1 was highly expressed in anaplastic Salivary cancer thyroid carcinomas (Sadow et al., 2008). Note Cholangiocarcinoma Using 2-dimensional differential in-gel Note electrophoresis, increased Stathmin 1 expression In a cohort of 80 extrahepatic cholangiocarcinoma was found in adenoid cystic carcinoma (Nakashima patients, high levels of Stathmin 1 correlated with et al., 2006). invasion and shorter recurrence-free survival by multivariate analysis (Watanabe et al., 2014). The To be noted authors also demonstrated that Stathmin 1 silencing Note resulted in reduced cell proliferation capacity and Although Stathmin 1 regulates multiple important increased sensitivity to paclitaxel treatment in a cellular functions, Schubart and colleagues cholangiocarcinoma cell line (Watanabe et al., (Schubart et al., 1996) initially reported that Stmn1 2014). knockout mice had normal growth, development, Melanoma reproduction and did not show any aberrant Note phenotype. Later on, it was observed that Stmn1 In primary melanoma samples, Stathmin 1 was knockout mice developed an axonopathy of the highly expressed in two independent cohorts, but central and peripheral nervous systems with aging did not impacts survival (Chen et al., 2013a). (Liedtke et al., 2002). In addition, Shumyatsky and Stathmin 1 silencing reduced cell proliferation and colleagues (Shumyatsky et al., 2005) reported that migration. Furthermore, Stathmin 1 overexpression Stmn1 knockout mice had reduced memory in potentiated both these cell processes in melanoma amygdala-dependent fear conditioning, failed to cell lines (Malme-3M and A375 cells) (Chen et al., recognize danger environments (Shumyatsky et al., 2013a). 2005) and exhibited accelerated fear extinction (Martel et al., 2012). Using well-defined mouse Mesothelioma models of carcinogenesis and Stmn1 knockout Note mice, D'Andrea and colleagues (D'Andrea et al., Overexpression of Stathmin 1 was found in all 2012) demonstrated that Stmn1 did not impact on mesothelioma cell lines tested (LRK1A, H2052, cancer onset. Regarding hematopoietic-related 211H, H290, MS1, H513 and H28 cells) and also in processes, Stmn1 knockout mice presented two primary tumors compared to its matched normal human hematopoietic disease phenotypes: tissue (Kim et al., 2007). megaloblastic anemia and thrombocytosis (Ramlogan-Steel et al., 2012). Notably, Stmn1 Pediatric brain cancer knockout mice did not have any that were Note alterations incompatible with life, and Stathmin 1 Using 2-dimensional differential in-gel inhibition in several types of cancer cells reduced electrophoresis, immunohistochemistry and the malignant phenotype, making it an attractive quantitative PCR, Stathmin 1 was identified as target for anticancer therapies.

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References tumours. Br J Cancer. 2000 Jan;82(1):142-50 Price DK, Ball JR, Bahrani-Mostafavi Z, Vachris JC, Feuerstein N, Cooper HL. Rapid protein phosphorylation Kaufman JS, Naumann RW, Higgins RV, Hall JB. The induced by phorbol ester in HL-60 cells. Unique alkali- phosphoprotein Op18/stathmin is differentially expressed stable phosphorylation of a 17,000-dalton protein detected in ovarian cancer. Cancer Invest. 2000;18(8):722-30 by two-dimensional gel electrophoresis. J Biol Chem. 1983 Iancu C, Mistry SJ, Arkin S, Wallenstein S, Atweh GF. Sep 10;258(17):10786-93 Effects of stathmin inhibition on the mitotic spindle. J Cell Hanash SM, Strahler JR, Kuick R, Chu EH, Nichols D. Sci. 2001 Mar;114(Pt 5):909-16 Identification of a polypeptide associated with the Liedtke W, Leman EE, Fyffe RE, Raine CS, Schubart UK. malignant phenotype in acute leukemia. J Biol Chem. 1988 Stathmin-deficient mice develop an age-dependent Sep 15;263(26):12813-5 axonopathy of the central and peripheral nervous systems. Melhem RF, Zhu XX, Hailat N, Strahler JR, Hanash SM. Am J Pathol. 2002 Feb;160(2):469-80 Characterization of the gene for a proliferation-related Misek DE, Chang CL, Kuick R, Hinderer R, Giordano TJ, phosphoprotein (oncoprotein 18) expressed in high Beer DG, Hanash SM. Transforming properties of a Q18-- amounts in acute leukemia. J Biol Chem. 1991 Sep >E mutation of the microtubule regulator Op18. Cancer 25;266(27):17747-53 Cell. 2002 Sep;2(3):217-28 Guy GR, Cao X, Chua SP, Tan YH. Okadaic acid mimics Takahashi M, Yang XJ, Lavery TT, Furge KA, Williams multiple changes in early protein phosphorylation and gene BO, Tretiakova M, Montag A, Vogelzang NJ, Re GG, expression induced by tumor necrosis factor or interleukin- Garvin AJ, Söderhäll S, Kagawa S, Hazel-Martin D, 1. J Biol Chem. 1992 Jan 25;267(3):1846-52 Nordenskjold A, Teh BT. Gene expression profiling of Brattsand G, Roos G, Marklund U, Ueda H, Landberg G, favorable histology Wilms tumors and its correlation with Nånberg E, Sideras P, Gullberg M. Quantitative analysis of clinical features. Cancer Res. 2002 Nov 15;62(22):6598- the expression and regulation of an activation-regulated 605 phosphoprotein (oncoprotein 18) in normal and neoplastic Chen G, Wang H, Gharib TG, Huang CC, Thomas DG, cells. Leukemia. 1993 Apr;7(4):569-79 Shedden KA, Kuick R, Taylor JM, Kardia SL, Misek DE, Roos G, Brattsand G, Landberg G, Marklund U, Gullberg Giordano TJ, Iannettoni MD, Orringer MB, Hanash SM, M. Expression of oncoprotein 18 in human leukemias and Beer DG. Overexpression of oncoprotein 18 correlates lymphomas. Leukemia. 1993 Oct;7(10):1538-46 with poor differentiation in lung adenocarcinomas. Mol Cell Proteomics. 2003 Feb;2(2):107-16 Jeha S, Luo XN, Beran M, Kantarjian H, Atweh GF. Antisense RNA inhibition of phosphoprotein p18 Rosell R, Scagliotti G, Danenberg KD, Lord RV, Bepler G, expression abrogates the transformed phenotype of Novello S, Cooc J, Crinò L, Sánchez JJ, Taron M, Boni C, leukemic cells. Cancer Res. 1996 Mar 15;56(6):1445-50 De Marinis F, Tonato M, Marangolo M, Gozzelino F, Di Costanzo F, Rinaldi M, Salonga D, Stephens C. Schubart UK, Yu J, Amat JA, Wang Z, Hoffmann MK, Transcripts in pretreatment biopsies from a three-arm Edelmann W. Normal development of mice lacking randomized trial in metastatic non-small-cell lung cancer. metablastin (P19), a phosphoprotein implicated in cell Oncogene. 2003 Jun 5;22(23):3548-53 cycle regulation. J Biol Chem. 1996 Jun 14;271(24):14062- 6 Neben K, Korshunov A, Benner A, Wrobel G, Hahn M, Kokocinski F, Golanov A, Joos S, Lichter P. Microarray- Alaiya AA, Franzén B, Fujioka K, Moberger B, Schedvins based screening for molecular markers in K, Silfversvärd C, Linder S, Auer G. Phenotypic analysis of medulloblastoma revealed STK15 as independent ovarian carcinoma: polypeptide expression in benign, predictor for survival. Cancer Res. 2004 May 1;64(9):3103- borderline and malignant tumors. Int J Cancer. 1997 Nov 11 27;73(5):678-83 Onda M, Emi M, Yoshida A, Miyamoto S, Akaishi J, Asaka Melhem R, Hailat N, Kuick R, Hanash SM. Quantitative S, Mizutani K, Shimizu K, Nagahama M, Ito K, Tanaka T, analysis of Op18 phosphorylation in childhood acute Tsunoda T. Comprehensive gene expression profiling of leukemia. Leukemia. 1997 Oct;11(10):1690-5 anaplastic thyroid cancers with cDNA microarray of 25 344 genes. Endocr Relat Cancer. 2004 Dec;11(4):843-54 Tournebize R, Andersen SS, Verde F, Dorée M, Karsenti E, Hyman AA. Distinct roles of PP1 and PP2A-like Koike H, Uzawa K, Nakashima D, Shimada K, Kato Y, phosphatases in control of microtubule dynamics during Higo M, Kouzu Y, Endo Y, Kasamatsu A, Tanzawa H. mitosis. EMBO J. 1997 Sep 15;16(18):5537-49 Identification of differentially expressed proteins in oral squamous cell carcinoma using a global proteomic Mistry SJ, Li HC, Atweh GF. Role for protein phosphatases approach. Int J Oncol. 2005 Jul;27(1):59-67 in the cell-cycle-regulated phosphorylation of stathmin. Biochem J. 1998 Aug 15;334 ( Pt 1):23-9 Mistry SJ, Bank A, Atweh GF. Targeting stathmin in prostate cancer. Mol Cancer Ther. 2005 Dec;4(12):1821-9 Curmi PA, Gavet O, Charbaut E, Ozon S, Lachkar- Colmerauer S, Manceau V, Siavoshian S, Maucuer A, Shumyatsky GP, Malleret G, Shin RM, Takizawa S, Tully Sobel A. Stathmin and its phosphoprotein family: general K, Tsvetkov E, Zakharenko SS, Joseph J, Vronskaya S, properties, biochemical and functional interaction with Yin D, Schubart UK, Kandel ER, Bolshakov VY. stathmin, tubulin. Cell Struct Funct. 1999 Oct;24(5):345-57 a gene enriched in the amygdala, controls both learned and innate fear. Cell. 2005 Nov 18;123(4):697-709 Brattsand G. Correlation of oncoprotein 18/stathmin expression in human breast cancer with established Varambally S, Yu J, Laxman B, Rhodes DR, Mehra R, prognostic factors. Br J Cancer. 2000 Aug;83(3):311-8 Tomlins SA, Shah RB, Chandran U, Monzon FA, Becich MJ, Wei JT, Pienta KJ, Ghosh D, Rubin MA, Chinnaiyan Curmi PA, Noguès C, Lachkar S, Carelle N, Gonthier MP, AM. Integrative genomic and proteomic analysis of Sobel A, Lidereau R, Bièche I. Overexpression of stathmin prostate cancer reveals signatures of metastatic in breast carcinomas points out to highly proliferative progression. Cancer Cell. 2005 Nov;8(5):393-406

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 133 STMN1 (stathmin 1) Machado-Neto JA, Traina F

Holmfeldt P, Brännström K, Stenmark S, Gullberg M. influences sarcoma cell shape, motility, and metastatic Aneugenic activity of Op18/stathmin is potentiated by the potential. Mol Biol Cell. 2008 May;19(5):2003-13 somatic Q18-->e mutation in leukemic cells. Mol Biol Cell. 2006 Jul;17(7):2921-30 Cheng AL, Huang WG, Chen ZC, Peng F, Zhang PF, Li MY, Li F, Li JL, Li C, Yi H, Yi B, Xiao ZQ. Identification of Kouzu Y, Uzawa K, Koike H, Saito K, Nakashima D, Higo novel nasopharyngeal carcinoma biomarkers by laser M, Endo Y, Kasamatsu A, Shiiba M, Bukawa H, Yokoe H, capture microdissection and proteomic analysis. Clin Tanzawa H. Overexpression of stathmin in oral squamous- Cancer Res. 2008 Jan 15;14(2):435-45 cell carcinoma: correlation with tumour progression and poor prognosis. Br J Cancer. 2006 Mar 13;94(5):717-23 Decaux O, Lodé L, Magrangeas F, Charbonnel C, Gouraud W, Jézéquel P, Attal M, Harousseau JL, Moreau Nakashima D, Uzawa K, Kasamatsu A, Koike H, Endo Y, P, Bataille R, Campion L, Avet-Loiseau H, Minvielle S. Saito K, Hashitani S, Numata T, Urade M, Tanzawa H. Prediction of survival in multiple myeloma based on gene Protein expression profiling identifies maspin and stathmin expression profiles reveals cell cycle and chromosomal as potential biomarkers of adenoid cystic carcinoma of the instability signatures in high-risk patients and hyperdiploid salivary glands. Int J Cancer. 2006 Feb 1;118(3):704-13 signatures in low-risk patients: a study of the Intergroupe Francophone du Myélome. J Clin Oncol. 2008 Oct Yuan RH, Jeng YM, Chen HL, Lai PL, Pan HW, Hsieh FJ, 10;26(29):4798-805 Lin CY, Lee PH, Hsu HC. Stathmin overexpression cooperates with p53 mutation and osteopontin Golouh R, Cufer T, Sadikov A, Nussdorfer P, Usher PA, overexpression, and is associated with tumour Brünner N, Schmitt M, Lesche R, Maier S, Timmermans M, progression, early recurrence, and poor prognosis in Foekens JA, Martens JW. The prognostic value of hepatocellular carcinoma. J Pathol. 2006 Aug;209(4):549- Stathmin-1, S100A2, and SYK proteins in ER-positive 58 primary breast cancer patients treated with adjuvant tamoxifen monotherapy: an immunohistochemical study. de Bont JM, den Boer ML, Kros JM, Passier MM, Breast Cancer Res Treat. 2008 Jul;110(2):317-26 Reddingius RE, Smitt PA, Luider TM, Pieters R. Identification of novel biomarkers in pediatric primitive Jiang H, Schiffer E, Song Z, Wang J, Zürbig P, Thedieck neuroectodermal tumors and ependymomas by proteome- K, Moes S, Bantel H, Saal N, Jantos J, Brecht M, Jenö P, wide analysis. J Neuropathol Exp Neurol. 2007 Hall MN, Hager K, Manns MP, Hecker H, Ganser A, Jun;66(6):505-16 Döhner K, Bartke A, Meissner C, Mischak H, Ju Z, Rudolph KL. Proteins induced by telomere dysfunction and Ghosh R, Gu G, Tillman E, Yuan J, Wang Y, Fazli L, DNA damage represent biomarkers of human aging and Rennie PS, Kasper S. Increased expression and disease. Proc Natl Acad Sci U S A. 2008 Aug differential phosphorylation of stathmin may promote 12;105(32):11299-304 prostate cancer progression. Prostate. 2007 Jul 1;67(10):1038-52 Rana S, Maples PB, Senzer N, Nemunaitis J. Stathmin 1: a novel therapeutic target for anticancer activity. Expert Kim JY, Harvard C, You L, Xu Z, Kuchenbecker K, Rev Anticancer Ther. 2008 Sep;8(9):1461-70 Baehner R, Jablons D. Stathmin is overexpressed in malignant mesothelioma. Anticancer Res. 2007 Jan- Sadow PM, Rumilla KM, Erickson LA, Lloyd RV. Stathmin Feb;27(1A):39-44 expression in pheochromocytomas, paragangliomas, and in other endocrine tumors. Endocr Pathol. 2008 Ngo TT, Peng T, Liang XJ, Akeju O, Pastorino S, Zhang Summer;19(2):97-103 W, Kotliarov Y, Zenklusen JC, Fine HA, Maric D, Wen PY, De Girolami U, Black PM, Wu WW, Shen RF, Jeffries NO, Wei SH, Lin F, Wang X, Gao P, Zhang HZ. Prognostic Kang DW, Park JK. The 1p-encoded protein stathmin and significance of stathmin expression in correlation with resistance of malignant gliomas to nitrosoureas. J Natl metastasis and clinicopathological characteristics in Cancer Inst. 2007 Apr 18;99(8):639-52 human ovarian carcinoma. Acta Histochem. 2008;110(1):59-65 Oishi Y, Nagasaki K, Miyata S, Matsuura M, Nishimura S, Akiyama F, Iwai T, Miki Y. Functional pathway Aoki D, Oda Y, Hattori S, Taguchi K, Ohishi Y, Basaki Y, characterized by gene expression analysis of Oie S, Suzuki N, Kono S, Tsuneyoshi M, Ono M, supraclavicular lymph node metastasis-positive breast Yanagawa T, Kuwano M. Overexpression of class III beta- cancer. J Hum Genet. 2007;52(3):271-9 tubulin predicts good response to taxane-based chemotherapy in ovarian clear cell adenocarcinoma. Clin Saal LH, Johansson P, Holm K, Gruvberger-Saal SK, She Cancer Res. 2009 Feb 15;15(4):1473-80 QB, Maurer M, Koujak S, Ferrando AA, Malmström P, Memeo L, Isola J, Bendahl PO, Rosen N, Hibshoosh H, Jiang L, Chen Y, Chan CY, Wang X, Lin L, He ML, Lin MC, Ringnér M, Borg A, Parsons R. Poor prognosis in Yew DT, Sung JJ, Li JC, Kung HF. Down-regulation of carcinoma is associated with a gene expression signature stathmin is required for TGF-beta inducible early gene 1 of aberrant PTEN tumor suppressor pathway activity. Proc induced growth inhibition of pancreatic cancer cells. Natl Acad Sci U S A. 2007 May 1;104(18):7564-9 Cancer Lett. 2009 Feb 8;274(1):101-8 Sewell DA, Yuan CX, Robertson E. Proteomic signatures Kuo MF, Wang HS, Kuo QT, Shun CT, Hsu HC, Yang SH, in laryngeal squamous cell carcinoma. ORL J Yuan RH. High expression of stathmin protein predicts a Otorhinolaryngol Relat Spec. 2007;69(2):77-84 fulminant course in medulloblastoma. J Neurosurg Pediatr. 2009 Jul;4(1):74-80 Singer S, Ehemann V, Brauckhoff A, Keith M, Vreden S, Schirmacher P, Breuhahn K. Protumorigenic Lin WC, Chen SC, Hu FC, Chueh SC, Pu YS, Yu HJ, overexpression of stathmin/Op18 by gain-of-function Huang KH. Expression of stathmin in localized upper mutation in p53 in human hepatocarcinogenesis. urinary tract urothelial carcinoma: correlations with Hepatology. 2007 Sep;46(3):759-68 prognosis. Urology. 2009 Dec;74(6):1264-9 Belletti B, Nicoloso MS, Schiappacassi M, Berton S, Lovat Ogino S, Nosho K, Baba Y, Kure S, Shima K, Irahara N, F, Wolf K, Canzonieri V, D'Andrea S, Zucchetto A, Friedl Toyoda S, Chen L, Kirkner GJ, Wolpin BM, Chan AT, P, Colombatti A, Baldassarre G. Stathmin activity Giovannucci EL, Fuchs CS. A cohort study of STMN1

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expression in colorectal cancer: body mass index and Li N, Jiang P, Du W, Wu Z, Li C, Qiao M, Yang X, Wu M. prognosis. Am J Gastroenterol. 2009 Aug;104(8):2047-56 Siva1 suppresses epithelial-mesenchymal transition and metastasis of tumor cells by inhibiting stathmin and Salvesen HB, Carter SL, Mannelqvist M, Dutt A, Getz G, stabilizing microtubules. Proc Natl Acad Sci U S A. 2011 Stefansson IM, Raeder MB, Sos ML, Engelsen IB, Trovik Aug 2;108(31):12851-6 J, Wik E, Greulich H, Bø TH, Jonassen I, Thomas RK, Zander T, Garraway LA, Oyan AM, Sellers WR, Kalland Lin WC, Chen SC, Chuang YT, Kuo KL, Huang KH. KH, Meyerson M, Akslen LA, Beroukhim R. Integrated Stathmin immunoreactivity in phaeochromocytomas and genomic profiling of endometrial carcinoma associates paragangliomas: differential expression between benign aggressive tumors with indicators of PI3 kinase activation. and malignant neoplasms. Asian J Surg. 2011 Proc Natl Acad Sci U S A. 2009 Mar 24;106(12):4834-9 Jan;34(1):15-22 Singer S, Malz M, Herpel E, Warth A, Bissinger M, Keith Trovik J, Wik E, Stefansson IM, Marcickiewicz J, M, Muley T, Meister M, Hoffmann H, Penzel R, Gdynia G, Tingulstad S, Staff AC, Njolstad TS, Vandenput I, Amant F, Ehemann V, Schnabel PA, Kuner R, Huber P, Akslen LA, Salvesen HB. Stathmin overexpression Schirmacher P, Breuhahn K. Coordinated expression of identifies high-risk patients and lymph node metastasis in stathmin family members by far upstream sequence endometrial cancer. Clin Cancer Res. 2011 May element-binding protein-1 increases motility in non-small 15;17(10):3368-77 cell lung cancer. Cancer Res. 2009 Mar 15;69(6):2234-43 Wosnitzer MS, Domingo-Domenech J, Castillo-Martin M, Su D, Smith SM, Preti M, Schwartz P, Rutherford TJ, Ritch C, Mansukhani M, Petrylack DP, Benson MC, Menato G, Danese S, Ma S, Yu H, Katsaros D. Stathmin McKiernan JM, Cordon-Cardo C. Predictive value of and tubulin expression and survival of ovarian cancer microtubule associated proteins tau and stathmin in patients receiving platinum treatment with and without patients with nonmuscle invasive bladder cancer receiving paclitaxel. Cancer. 2009 Jun 1;115(11):2453-63 adjuvant intravesical taxane therapy. J Urol. 2011 Nov;186(5):2094-100 Xi W, Rui W, Fang L, Ke D, Ping G, Hui-Zhong Z. Expression of stathmin/op18 as a significant prognostic Baquero MT, Hanna JA, Neumeister V, Cheng H, Molinaro factor for cervical carcinoma patients. J Cancer Res Clin AM, Harris LN, Rimm DL. Stathmin expression and its Oncol. 2009 Jun;135(6):837-46 relationship to microtubule-associated protein tau and outcome in breast cancer. Cancer. 2012 Oct Björklund P, Cupisti K, Fryknäs M, Isaksson A, Willenberg 1;118(19):4660-9 HS, Akerström G, Hellman P, Westin G. Stathmin as a marker for malignancy in pheochromocytomas. Exp Clin Bhagirath D, Abrol N, Khan R, Sharma M, Seth A, Sharma Endocrinol Diabetes. 2010 Jan;118(1):27-30 A. Expression of CD147, BIGH3 and Stathmin and their potential role as diagnostic marker in patients with Gan L, Guo K, Li Y, Kang X, Sun L, Shu H, Liu Y. Up- urothelial carcinoma of the bladder. Clin Chim Acta. 2012 regulated expression of stathmin may be associated with Oct 9;413(19-20):1641-6 hepatocarcinogenesis. Oncol Rep. 2010 Apr;23(4):1037- 43 Canzonieri V, Barzan L, Franchin G, Vaccher E, Talamini R, Sulfaro S, Baldassarre G. Alteration of G1/S transition Hsieh SY, Huang SF, Yu MC, Yeh TS, Chen TC, Lin YJ, regulators influences recurrences in head and neck Chang CJ, Sung CM, Lee YL, Hsu CY. Stathmin1 squamous carcinomas. J Cell Physiol. 2012 overexpression associated with polyploidy, tumor-cell Jan;227(1):233-8 invasion, early recurrence, and poor prognosis in human hepatoma. Mol Carcinog. 2010 May;49(5):476-87 D'Andrea S, Berton S, Segatto I, Fabris L, Canzonieri V, Colombatti A, Vecchione A, Belletti B, Baldassarre G. Jeon TY, Han ME, Lee YW, Lee YS, Kim GH, Song GA, Stathmin is dispensable for tumor onset in mice. PLoS Hur GY, Kim JY, Kim HJ, Yoon S, Baek SY, Kim BS, Kim One. 2012;7(9):e45561 JB, Oh SO. Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and Dong B, Mu L, Qin X, Qiao W, Liu X, Yang L, Xue L, migration of gastric cancer cells. Br J Cancer. 2010 Feb Rainov NG, Liu X. Stathmin expression in glioma-derived 16;102(4):710-8 microvascular endothelial cells: a novel therapeutic target. Oncol Rep. 2012 Mar;27(3):714-8 Trovik J, Wik E, Stefansson I, Carter SL, Beroukhim R, Oyan AM, Kalland KH, Akslen LA, Salvesen HB. Stathmin Dubosq F, Ploussard G, Soliman H, Turpin E, Latil A, is superior to AKT and phospho-AKT staining for the Desgrandchamps F, de The H, Mongiat-Artus P. detection of phosphoinositide 3-kinase activation and Identification of a three-gene expression signature of early aggressive endometrial cancer. Histopathology. 2010 recurrence in non-muscle-invasive urothelial cell Oct;57(4):641-6 carcinoma of the bladder. Urol Oncol. 2012 Nov- Dec;30(6):833-40 Zheng P, Liu YX, Chen L, Liu XH, Xiao ZQ, Zhao L, Li GQ, Zhou J, Ding YQ, Li JM. Stathmin, a new target of PRL-3 Kang W, Tong JH, Chan AW, Lung RW, Chau SL, Wong identified by proteomic methods, plays a key role in QW, Wong N, Yu J, Cheng AS, To KF. Stathmin1 plays progression and metastasis of colorectal cancer. J oncogenic role and is a target of microRNA-223 in gastric Proteome Res. 2010 Oct 1;9(10):4897-905 cancer. PLoS One. 2012;7(3):e33919 Belletti B, Baldassarre G. Stathmin: a protein with many Martel G, Hevi C, Wong A, Zushida K, Uchida S, tasks. New biomarker and potential target in cancer. Shumyatsky GP. Murine GRPR and stathmin control in Expert Opin Ther Targets. 2011 Nov;15(11):1249-66 opposite directions both cued fear extinction and neural activities of the amygdala and prefrontal cortex. PLoS One. Karst AM, Levanon K, Duraisamy S, Liu JF, Hirsch MS, 2012;7(2):e30942 Hecht JL, Drapkin R. Stathmin 1, a marker of PI3K pathway activation and regulator of microtubule dynamics, Ramlogan-Steel CA, Steel JC, Fathallah H, Iancu-Rubin C, is expressed in early pelvic serous carcinomas. Gynecol Soleimani M, Dong, Atweh GF.. The Role of Stathmin, a Oncol. 2011 Oct;123(1):5-12 Regulator of Mitosis, in Hematopoiesis. Blood (ASH

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Annual Meeting Abstracts). 2012 120(21): Abstract #3453. Barbareschi M, Valeri B, Canevari S, Colombatti A, Belletti B, Del Sal G, Baldassarre G.. Stathmin regulates mutant Tan HT, Wu W, Ng YZ, Zhang X, Yan B, Ong CW, Tan S, p53 stability and transcriptional activity in ovarian cancer. Salto-Tellez M, Hooi SC, Chung MC.. Proteomic analysis EMBO Mol Med. 2013 May;5(5):707-22. doi: of colorectal cancer metastasis: stathmin-1 revealed as a 10.1002/emmm.201201504. Epub 2013 Apr 22. player in cancer cell migration and prognostic marker. J Proteome Res. 2012 Feb 3;11(2):1433-45. doi: Wik E, Birkeland E, Trovik J, Werner HM, Hoivik EA, Mjos 10.1021/pr2010956. Epub 2012 Jan 10. S, Krakstad C, Kusonmano K, Mauland K, Stefansson IM, Holst F, Petersen K, Oyan AM, Simon R, Kalland KH, Williams K, Ghosh R, Giridhar PV, Gu G, Case T, Belcher Ricketts W, Akslen LA, Salvesen HB.. High phospho- SM, Kasper S.. Inhibition of stathmin1 accelerates the Stathmin(Serine38) expression identifies aggressive metastatic process. Cancer Res. 2012 Oct endometrial cancer and suggests an association with PI3K 15;72(20):5407-17. doi: 10.1158/0008-5472.CAN-12-1158. inhibition. Clin Cancer Res. 2013 May 1;19(9):2331-41. Epub 2012 Aug 21. doi: 10.1158/1078-0432.CCR-12-3413. Epub 2013 Mar 28. Akhtar J, Wang Z, Zhang ZP, Bi MM.. Lentiviral-mediated Hsu HP, Li CF, Lee SW, Wu WR, Chen TJ, Chang KY, RNA interference targeting stathmin1 gene in human Liang SS, Tsai CJ, Shiue YL.. Overexpression of stathmin gastric cancer cells inhibits proliferation in vitro and tumor 1 confers an independent prognostic indicator in growth in vivo. J Transl Med. 2013 Sep 16;11:212. doi: nasopharyngeal carcinoma. Tumour Biol. 2014 10.1186/1479-5876-11-212. Mar;35(3):2619-29. doi: 10.1007/s13277-013-1345-3. Chen J, Abi-Daoud M, Wang A, Yang X, Zhang X, Feilotter Epub 2013 Nov 12. HE, Tron VA.. Stathmin 1 is a potential novel oncogene in Lu Y, Liu C, Cheng H, Xu Y, Jiang J, Xu J, Long J, Liu L, melanoma. Oncogene. 2013a Mar 7;32(10):1330-7. doi: Yu X.. Stathmin, interacting with Nf-kB, promotes tumor 10.1038/onc.2012.141. Epub 2012 Jun 4. growth and predicts poor prognosis of pancreatic cancer. Chen YL, Uen YH, Li CF, Horng KC, Chen LR, Wu WR, Curr Mol Med. 2014 Mar;14(3):328-39. Tseng HY, Huang HY, Wu LC, Shiue YL.. The E2F Machado-Neto JA, de Melo Campos P, Favaro P, Lazarini transcription factor 1 transactives stathmin 1 in M, Lorand-Metze I, Costa FF, Olalla Saad ST, Traina F.. hepatocellular carcinoma. Ann Surg Oncol. 2013b Stathmin 1 is involved in the highly proliferative phenotype Nov;20(12):4041-54. doi: 10.1245/s10434-012-2519-8. of high-risk myelodysplastic syndromes and acute Epub 2012 Aug 22. leukemia cells. Leuk Res. 2014a Feb;38(2):251-7. doi: Howitt BE, Nucci MR, Drapkin R, Crum CP, Hirsch MS.. 10.1016/j.leukres.2013.11.013. Epub 2013 Dec 1. Stathmin-1 expression as a complement to p16 helps Machado-Neto JA, Saad ST, Traina F.. Stathmin 1 in identify high-grade cervical intraepithelial neoplasia with normal and malignant hematopoiesis. BMB Rep. 2014b increased specificity. Am J Surg Pathol. 2013 Mar 18. pii: 2687. [Epub ahead of print] Jan;37(1):89-97. doi: 10.1097/PAS.0b013e3182753f5a. Watanabe A, Suzuki H, Yokobori T, Tsukagoshi M, Altan Ke B, Wu LL, Liu N, Zhang RP, Wang CL, Liang H.. B, Kubo N, Suzuki S, Araki K, Wada S, Kashiwabara K, Overexpression of stathmin 1 is associated with poor Hosouchi Y, Kuwano H.. Stathmin1 regulates p27 prognosis of patients with gastric cancer. Tumour Biol. expression, proliferation and drug resistance, resulting in 2013 Oct;34(5):3137-45. doi: 10.1007/s13277-013-0882-0. poor clinical prognosis in cholangiocarcinoma. Cancer Sci. Epub 2013 Jun 13. 2014 Jun;105(6):690-6. doi: 10.1111/cas.12417. Epub Marafioti T, Copie-Bergman C, Calaminici M, Paterson JC, 2014 May 27. Shende VH, Liu H, Baia M, Ramsay AD, Agostinelli C, Werner HM, Trovik J, Halle MK, Wik E, Akslen LA, Briere J, Clear A, Du MQ, Piccaluga PP, Masir N, Nacheva Birkeland E, Bredholt T, Tangen IL, Krakstad C, Salvesen EP, Sujobert P, Shanmugam K, Grogan TM, Brooks SP, HB.. Stathmin protein level, a potential predictive marker Khwaja A, Ardeshna K, Townsend W, Pileri SA, Haioun C, for taxane treatment response in endometrial cancer. Linch D, Gribben JG, Gaulard P, Isaacson PG.. Another PLoS One. 2014 Feb 25;9(2):e90141. doi: look at follicular lymphoma: immunophenotypic and 10.1371/journal.pone.0090141. eCollection 2014. molecular analyses identify distinct follicular lymphoma subgroups. Histopathology. 2013 May;62(6):860-75. doi: Wu Y, Tang M, Wu Y, Weng X, Yang L, Xu W, Yi W, Gao 10.1111/his.12076. Epub 2013 Mar 20. J, Bode AM, Dong Z, Cao Y.. A combination of paclitaxel and siRNA-mediated silencing of Stathmin inhibits growth Miceli C, Tejada A, Castaneda A, Mistry SJ.. Cell cycle and promotes apoptosis of nasopharyngeal carcinoma inhibition therapy that targets stathmin in in vitro and in cells. Cell Oncol (Dordr). 2014 Feb;37(1):53-67. doi: vivo models of breast cancer. Cancer Gene Ther. 2013 10.1007/s13402-013-0163-3. Epub 2013 Dec 5. May;20(5):298-307. doi: 10.1038/cgt.2013.21. Epub 2013 Apr 26. This article should be referenced as such: Sonego M, Schiappacassi M, Lovisa S, Dall'Acqua A, Machado-Neto JA, Traina F. STMN1 (stathmin 1). Atlas Bagnoli M, Lovat F, Libra M, D'Andrea S, Canzonieri V, Genet Cytogenet Oncol Haematol. 2015; 19(2):126-136. Militello L, Napoli M, Giorda G, Pivetta B, Mezzanzanica D,

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Gene Section Review

MTA3 (metastasis associated 1 family, member 3 ) Ansgar Brüning, Ioannis Mylonas University Hospital Munich, Department of Obstetrics/Gynaecology, Molecular Biology Laboratory, Marchioninistrasse 15, 81377 Munchen, Germany (AB, IM) Published in Atlas Database: June 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/MTA3ID41445ch2p21.html DOI: 10.4267/2042/56299 This article is an update of : Brüning A, Mylonas I. MTA3 (metastasis associated 1 family, member 3). Atlas Genet Cytogenet Oncol Haematol 2011;15(7):567-569.

Abstract development; cell cycle MTA3 functions as a transcriptional repressor Identity through interacting with histone deacetylases and Other names: KIAA1266 nucleosome remodelling complexes such as Mi- 2/NuRD. HGNC (Hugo): MTA3 Since MTA3 inhibits expression of SNAIL, a Location: 2p21 transcriptional repressor of the cell adhesion protein E-cadherin, downregulation of MTA3 was found to DNA/RNA be associated with reduced E-cadherin levels and advanced cancer stages. Recent data also revealed Description upregulation of MTA3 in non-small lung cancer The human MTA3 gene was identified through cells and highly aggressive subtypes of endometrial sequence homologies to other members of the MTA cancer, additionally indicating cancer-promoting gene family (human MTA1, human MTA2, murine effects of MTA3. MTA3). Keywords The human MTA3 gene is composed of 14 exons. Metastasis; gene regulation; chromosome The MTA3 promoter sequence contains SP1, AP1, remodelling; oestrogen receptor; E-cadherin; B cell and oestrogen receptor binding sites (ER half sites).

Genomic organization of the human MTA3 gene. The intron/exon structure of MTA3 with start (ATG) and stop (TAA) codons indicated. All 14 exons are depicted; the intron sequences shortened for better graphical visualization.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 137 MTA3 (metastasis associated 1 family, member 3 ) Brüning A, Mylonas I

Domain structure of the MTA3 protein. BAH (bromo-adjacent homology) domain: putative protein-protein interaction domain, involved in gene silencing; ELM (Egl-27 and MTA1 homology) domain: unknown function; SANT (SWI3, ADA2, N-CoR and TFIIIB B) domain: putative DNA binding domain; ZnF (GATA-type zinc finger) domain: direct DNA binding domain.

Transcription endometrial cancer cells and cancer cell lines, in trophoblast cells and chorionic cancer cell lines, in Two open reading frames of 1785 bp (isoform 1; germinal centre B cells, and in B cell-derived 594 aa; MTA3L) and 1548 bp (isoform 2; 515 aa; lymphomas. MTA3S, MTA3) were identified and predicted to A tissue distribution analysis of MTA3 expression be transcribed. The smaller isoform (MTA3S = in mice revealed a widespread distribution of MTA3) appears to be the most abundantly MTA3 in the developing embryo and in adult expressed isoform at the RNA and protein level. tissues (heart, brain, spleen, lung, liver and kidney). Pseudogene Localisation PGO.9606.51655; PGO.9606.72237. MTA3 exhibits primarily a nuclear localisation, Protein although additional cytoplasmic localisation has been described. Description Function MTA3 functions as a transcriptional repressor by interacting with histone deacetylases and In epithelial cells, MTA3 maintains the expression nucleosome remodelling complexes such as Mi- of E-cadherin through the suppression of the E- 2/NuRD. cadherin inhibitor SNAIL. Expression of MTA3 is regulated by oestrogens via Expression direct binding of the oestrogen receptor to the MTA3 expression has been found in normal human MTA3 promoter and is thus involved in the breast, ovarian, lung, and endometrial epithelial generation and maintenance of oestrogen-dependent cells, in malignant breast, ovarian, lung, and epithelia such as the breast ductal epithelium.

The MTA3 regulation network. A. Breast ductal epithelia cells; epithelial cancer cells. B. Germinal center B lymphocytes; B cell-derived lymphomas. The regulation of MTA3 expression and its target genes by transcriptional activators (green) and transcriptional repressors (red) is shown. ER: oestrogen receptor.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 138 MTA3 (metastasis associated 1 family, member 3 ) Brüning A, Mylonas I

Mammary gland development suggested involvement of MTA3 in placenta Animal experiments revealed involvement of development and homeostasis. MTA3 expression MTA3 expression in mammary gland was found to be elevated in preeclampsia patients morphogenesis mediated by the suppression of the and to negatively interfere with hCG (human Wnt4 signalling pathway and upregulation of chorionic gonadotropin) expression. epithelial cell adhesion proteins such as E-cadherin. Homology Normal mammary gland development, as confirmed and studied by several knock out and MTA3 exhibits a high homology to human MTA1, knock in mouse models, relies on the concerted and MTA2, and murine MTA3. correct integration of divers signalling pathways, including the Wnt signalling pathway. Secretion of Implicated in Wnt factors and their binding by mammary epithelial cells is necessary for correct gland Endometrial cancer development and its deregulation has been Note described to be involved in tumorigenesis. MTA3 MTA3 expression is significantly reduced in has been shown to inhibit Wnt4 expression by its endometrioid adenocarcinomas of poor transcriptional repression function, causing reduced differentiation, although not associated with Wnt4 secretion and subsequent lower beta-catenin patients' survival. levels. Therefore, based on the observations made By contrast, in uterine non-endometrioid with transgenic mouse models, expression of carcinomas, a highly aggressive subtype of MTA3 in mammary epithelial cells has been endometrial cancer, MTA3 expression associated with the inhibition of ductal branching in demonstrated a significant association with FIGO virgin and pregnant murine mammary glands. surgical stage, lymph node involvement, and Epithelial cancer lymphovascular space invasion. Reduced MTA3 expression in epithelial breast In uterine non-endometroid cancer, MTA3 was also cancer, endometrial cancer, and ovarian cancer is revealed to be a significant independent prognostic associated with cancer progression by promoting parameter, associated with patients' progression- the epithelial-mesenchymal transition (EMT). It is free survival, cause-specific survival, and overall principally believed that reduced expression of survival. MTA3 allows higher expression levels of SNAIL Ovarian cancer and SLUG, two repressors of metastasis-associated cell adhesion proteins such as E-cadherin and Note occludin. Conversely, MTA3 expression was found MTA3 expression is reduced in ovarian cancer with to be elevated in non-small lung cancer and in poor differentiation, although not at significant uterine non-endometrioid cancer, in which a levels. growth-promoting effect of MTA3 overexpression Breast cancer was suggested. Note Haemangiogenesis and lymphomagenesis Although extensively studied on breast cancer cells A high expression level of MTA3 was found in and tissues, revealing a close correlation of MTA3 germinal centre B lymphocytes, suggesting an expression with oestrogen receptor expression, no involvement in B cell maturation by direct studies have yet shown a direct association of interaction with BCL6. BCL6 (B-cell lymphoma-6) MTA3 expression with clinicopathological is a transcriptional repressor that is co-expressed parameters in breast cancer. with MTA3 in the germinal centre, where normal B cells proliferate and undergo maturation. BCL6 Lung cancer functions as a transcriptional repressor and Note suppresses, in cooperation with MTA3, the MTA3 was found to be upregulated in NSCLC expression of PRDM1 (Pr domain-containing (non-small cell lung cancer) tissues and its protein 1), a master regulator of plasma cell overexpression correlated with lymph node status differentiation. Overexpression of BCL6 is often and poor patients' prognosis. observed in lymphomas, especially in large B-cell lymphomas. Thus, the cooperative action of BCL6 Gastric cancer, oesophagal cancer together with MTA3 is believed to block Note differentiation of large B-cell lymphomas, In gastroesophageal junction (GEJ) facilitating lymphomagenesis. adenocarcinomas, MTA3 expression was found to Placenta development be reduced in cancer tissues and reduced MTA3 A conspicuously high expression level of MTA3 in expression correlated with worse overall patients' trophoblast cells and trophoblast tumour cells has survival.

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Ovary (non-malignant) MTA3, is selectively expressed by germinal centre B cells and lymphomas of putative germinal centre derivation. J Note Pathol. 2007 Sep;213(1):106-15 Murine ovarian granulosa cells of all follicular Brüning A, Makovitzky J, Gingelmaier A, Friese K, Mylonas stages were shown to express high levels of MTA3. I. The metastasis-associated genes MTA1 and MTA3 are Expression of MTA3 in granulosa cells was also abundantly expressed in human placenta and chorionic shown to promote cell cycle progression by carcinoma cells. Histochem Cell Biol. 2009 Jul;132(1):33-8 controlling entry into the M phase. Li X, Jia S, Wang S, Wang Y, Meng A. Mta3-NuRD complex is a master regulator for initiation of primitive Placenta hematopoiesis in vertebrate embryos. Blood. 2009 Dec Note 24;114(27):5464-72 A high expression level of MTA3 was observed in Brüning A, Jückstock J, Blankenstein T, Makovitzky J, placental trophoblast cells and chorionic carcinoma Kunze S, Mylonas I. The metastasis-associated gene MTA3 is downregulated in advanced endometrioid cells. adenocarcinomas. Histol Histopathol. 2010 In patients with preeclamptic gestational disease, Nov;25(11):1447-56 MTA3 expression was found to be downregulated Liu X, Li X, Yin L, Ding J, Jin H, Feng Y. Genistein inhibits and shown to be involved in human chorionic placental choriocarcinoma cell line JAR invasion through gonadotropin expression by directly repressing the ER β/MTA3/Snail/E-cadherin pathway. Oncol Lett. 2011 hCG promoter, leading to dysregulated hCG Sep 1;2(5):891-897 expression as observed in preeclampsia. Kwintkiewicz J, Padilla-Banks E, Jefferson WN, Jacobs IM, Wade PA, Williams CJ. Metastasis-associated protein 3 (MTA3) regulates G2/M progression in proliferating mouse References granulosa cells. Biol Reprod. 2012 Mar;86(3):1-8 Fujita N, Jaye DL, Kajita M, Geigerman C, Moreno CS, Mylonas I, Brüning A. The metastasis-associated gene Wade PA. MTA3, a Mi-2/NuRD complex subunit, regulates MTA3 is an independent prognostic parameter in uterine an invasive growth pathway in breast cancer. Cell. 2003 non-endometrioid carcinomas. Histopathology. 2012 Apr 18;113(2):207-19 Mar;60(4):665-70 Fujita N, Jaye DL, Geigerman C, Akyildiz A, Mooney MR, Chen Y, Miyazaki J, Nishizawa H, Kurahashi H, Leach R, Boss JM, Wade PA. MTA3 and the Mi-2/NuRD complex Wang K. MTA3 regulates CGB5 and Snail genes in regulate cell fate during B lymphocyte differentiation. Cell. trophoblast. Biochem Biophys Res Commun. 2013 Apr 2004 Oct 1;119(1):75-86 19;433(4):379-84 Fujita N, Kajita M, Taysavang P, Wade PA. Hormonal Dong H, Guo H, Xie L, Wang G, Zhong X, Khoury T, Tan regulation of metastasis-associated protein 3 transcription D, Zhang H. The metastasis-associated gene MTA3, a in breast cancer cells. Mol Endocrinol. 2004 component of the Mi-2/NuRD transcriptional repression Dec;18(12):2937-49 complex, predicts prognosis of gastroesophageal junction Mishra SK, Talukder AH, Gururaj AE, Yang Z et al.. adenocarcinoma. PLoS One. 2013;8(5):e62986 Upstream determinants of estrogen receptor-alpha Li H, Sun L, Xu Y, Li Z, Luo W, Tang Z, Qiu X, Wang E. regulation of metastatic tumor antigen 3 pathway. J Biol Overexpression of MTA3 Correlates with Tumor Chem. 2004 Jul 30;279(31):32709-15 Progression in Non-Small Cell Lung Cancer. PLoS One. Zhang H, Singh RR, Talukder AH, Kumar R. Metastatic 2013;8(6):e66679 tumor antigen 3 is a direct corepressor of the Wnt4 Zheng S, Du Y, Chu H, Chen X, Li P, Wang Y, Ma Y, pathway. Genes Dev. 2006 Nov 1;20(21):2943-8 Wang H, Zang W, Zhang G, Zhao G. Analysis of MAT3 Zhang H, Stephens LC, Kumar R. Metastasis tumor gene expression in NSCLC. Diagn Pathol. 2013 Oct antigen family proteins during breast cancer progression 9;8:166 and metastasis in a reliable mouse model for human breast cancer. Clin Cancer Res. 2006 Mar 1;12(5):1479-86 This article should be referenced as such: Jaye DL, Iqbal J, Fujita N, Geigerman CM, Li S, Karanam Brüning A, Mylonas I. MTA3 (metastasis associated 1 S, Fu K, Weisenburger DD, Chan WC, Moreno CS, Wade family, member 3 ). Atlas Genet Cytogenet Oncol PA. The BCL6-associated transcriptional co-repressor, Haematol. 2015; 19(2):137-140.

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Leukaemia Section Short Communication t(10;11)(q22;q23) KMT2A/TET1 Antoine Ittel Laboratoire de Cytogenetique hematologique, CHU Hautepierre, Strasbourg, France (AI)

Published in Atlas Database: June 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t1011q22q23ID1410.html DOI: 10.4267/2042/56300 This article is an update of : Viguié F. t(10;11)(q22;q23). Atlas Genet Cytogenet Oncol Haematol 2009;13(2):137-138.

Abstract Cytogenetics Review on t(10;11)(q22;q23) KMT2A/TET1, with Cytogenetics morphological data on clinics, and the genes implicated. Easy to detect, evident 10q- and 11q+ derivatives. Clinics and pathology Cytogenetics molecular Disease Commercial dual color MLL/KMT2A FISH probes are splitted by the translocation. 10q22 breakpoint Acute myeloid leukemia (AML), B-cell precursor may be detected with RP11-119F7 BAC probe or acute lymphoblastic leukemia (ALL), T-cell using RP11-9E13 and RP11-314J18 BACs lymphoblastic lymphoma (separation of the two BACs with the Phenotype/cell stem origin t(10;11)(q22;q23)). Described in subtypes AML-M2, -M4 and -M5. Additional anomalies Cell lineage dysplasia may be associated. 5/11 caryotypes available have the t(10;11) as sole Described also in 2 B-ALL and one case of T-cell chromosomal abnormality. 4/11 have one more lymphoblastic lymphoma with a subsequent chromosomal abnormality (+21, del(6)p(21), transformation to AML. del(5)(q15), add(13)(p11) or Epidemiology der(6)t(6;?9)(p22;?q21) in 2 different clones with Described in 14 cases in the literature, mainly the t(10;11)); one case was hyperdiploid with 51 adults and 2 children (8 men, 5 women, 1 chromosomes; one case had complex caryotype. unknown) median age of 39 years (range 1 month to 67 years). Genes involved and Frequency estimated at 0.3 % of MLL-rearranged proteins acute leukemia cases (5 out 1590) (Lee et al., 2013). KMT2A Prognosis Location Undetermined, possibly intermediate. On the 8 of 11q23 14 patients with available clinical outcomes, 7 had Note a complete remission. KMT2A is also called MLL (mixed-lineage 6 patients died at an average of 16 months after leukemia or myeloid-lymphoid leukaemia), ALL-1 initial diagnosis. or HRX.

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A. t(10;11)(q22;q23) in RHG banding. B. The same with GTG banding.

A. FISH on metaphase (inverted DAPI) using RP11-9E13 and RP11-314J18 BACs showing the separation of the two probes in the t(10;11)(q22;q23). B. Focus on chromosomes 10 and 11 and derivative chromosomes (inverted DAPI) from the t(10;11) with the same BACs.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 142 t(10;11)(q22;q23) KMT2A/TET1 Ittel A

DNA/RNA (and the corresponding global increase of 5hmC) 36 exons, multiple transcripts 13-15 kb. cooperates with MLL fusions in orchestrating the transcriptional activation of their cotargets. The main critical oncogenic cotargets described are Protein HOXA9, MEIS1 and PBX3. 430 kDA, contains two DNA binding motifs (a AT The Hoxa9/Meis1/Pbx3 signaling cascade promotes hook and a CXXC domain), a DNA methyl cell proliferation and inhibits apoptosis/cell transferase motif, a bromodomain; transcriptional differentiation, thereby leading to cell regulatory factor involved in maintenance of Hox transformation and leukemogenesis. (Huang et al., gene expression during embryogenesis and during 2013). the process of haematopoietic progenitors expansion and differentiation. Result of the chromosomal TET1 anomaly Location 10q22 Hybrid gene Note Description TET1 is also called LCX (leukemia-associated Transcripts from the 5' MLL-LCX 3' fusion gene on protein with a CXXC domain) or CXXC6 (CXXC der(11) are expressed; transcripts from the 5' LCX- finger 6). MLL 3' counterpart are not detected. Breakpoints in the MLL gene are located between DNA/RNA intron 6 and exon 11. 8497 bp representing the whole coding sequence. All genomic breakpoints within the TET1 gene Contains 12 exons. Contains 3 bipartite nuclear were identified in an approximately 17 kb genomic localization sites, 1 alpha helice coiled-coil region region flanked by TET1 exons 8 and 12. and 1 cysteine rich domain with high level Most characterized breakpoints (5 of 7 cases) were homology with a CXXC DNA binding site. mapped to intron 8. Protein Fusion protein Predicted size of 2136 amino acids, expression restricted to some fetal tissues, mainly lung, heart Description and brain; not expressed in hematopoietic tissues, Predicted molecular weight of 204.4 kDa. except in spleen; unknown function. Oncogenesis TET family enzymes convert 5-methylcytosine Unknown; the alpha helice coiled-coil region (5mC) to 5-hydroxymethylcytosine (5hmC). Key retained at the COOH extremity might be involved role in active DNA demethylation. TET1 and in the leukemogenesis. TET2: key enzymes responsible for the presence of 5hmC in mouse embryonic stem cells (ESCs). References TET1: regulates the lineage differentiation potential of ESCs. TET1 interacts physically with NANOG, Aventín A, La Starza R, Martínez C, Wlodarska I, Boogaerts M, Van den Berghe H, Mecucci C. Involvement synergistically enhancing the efficiency of NANOG of MLL gene in a t(10;11)(q22;q23) and a t(8;11)(q24;q23) in somatic cell reprogramming. NANOG/TET1 co- identified by fluorescence in situ hybridization. Cancer occupy genomic loci of genes associated with both Genet Cytogenet. 1999 Jan 1;108(1):48-52 maintenance of pluripotency and lineage Ono R, Taki T, Taketani T, Taniwaki M, Kobayashi H, commitment in embryonic stem cells, and may Hayashi Y. LCX, leukemia-associated protein with a CXXC deposit 5hmC to target genes before the domain, is fused to MLL in acute myeloid leukemia with establishment of pluripotency. Taken together, trilineage dysplasia having t(10;11)(q22;q23). Cancer Res. 2002 Jul 15;62(14):4075-80 these observations suggest a possible mechanism for the lineage switch observed in one of the 14 Lorsbach RB, Moore J, Mathew S, Raimondi SC, Mukatira ST, Downing JR. TET1, a member of a novel protein cases. (Ittel et al., 2013). family, is fused to MLL in acute myeloid leukemia TET1 significantly up-regulated in MLL-rearranged containing the t(10;11)(q22;q23). Leukemia. 2003 leukemia. TET1: direct target gene of MLL-fusion Mar;17(3):637-41 proteins. MLL fusions bind to the promoter region Katoh M, Katoh M. Identification and characterization of of TET1 and promote its expression directly in both human CXXC10 gene in silico. Int J Oncol. 2004 human and mouse hematopoietic stem/progenitor Oct;25(4):1193-9 cells, cumulating in a global increase of 5hmC. Huang H, Jiang X, Li Z, Li Y, Song CX, He C, Sun M, Chen Briefly, MLL-fusion proteins bind directly to the P, Gurbuxani S, Wang J, Hong GM, Elkahloun AG, Tet1 locus. Consequently this promote its Arnovitz S, Wang J, Szulwach K, Lin L, Street C, expression and the increased expression of Tet1 Wunderlich M, Dawlaty M, Neilly MB, Jaenisch R, Yang FC, Mulloy JC, Jin P, Liu PP, Rowley JD, Xu M, He C,

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Chen J. TET1 plays an essential oncogenic role in MLL- Lee SG, Cho SY, Kim MJ, Oh SH, Cho EH, Lee S, Baek rearranged leukemia. Proc Natl Acad Sci U S A. 2013 Jul EJ, Choi JH, Bohlander SK, Lode L, Richebourg S, Yoon 16;110(29):11994-9 HJ, Marschalek R, Meyer C, Park TS. Genomic breakpoints and clinical features of MLL-TET1 Ittel A, Jeandidier E, Helias C, Perrusson N, Humbrecht C, rearrangement in acute leukemias. Haematologica. 2013 Lioure B, Mazurier I, Mayeur-Rousse C, Lavaux A, Apr;98(4):e55-7 Thiebault S, Lerintiu F, Gervais C, Mauvieux L. First description of the t(10;11)(q22;q23)/MLL-TET1 This article should be referenced as such: translocation in a T-cell lymphoblastic lymphoma, with subsequent lineage switch to acute myelomonocytic Ittel A. t(10;11)(q22;q23) KMT2A/TET1. Atlas Genet myeloid leukemia. Haematologica. 2013 Dec;98(12):e166- Cytogenet Oncol Haematol. 2015; 19(2):141-144. 8

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Leukaemia Section Short Communication t(2;9)(p23;q33) TRAF1/ALK Xiaoming Xing, Andrew L Feldman Department of Pathology, Affiliated Hospital of Medical College, Qingdao University, 16 Jiangsu Road, Qingdao, China (XX), Department of Laboratory Medicine and Pathology, College Of Medicine, Mayo Clinic, 200 First Street SW, Hilton Building, Room 8-00F, Rochester, MN 55905 USA (ALF)

Published in Atlas Database: June 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0209p23q33ID1685.html DOI: 10.4267/2042/56301 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract anthracycline-based multi-agent chemotherapy. Review on t(2;9)(p23;q33) TRAF1/ALK, with data Prognosis on clinics, and the genes implicated. Among peripheral T-cell lymphomas, ALK- positive ALCLs tend to have favorable outcomes. Clinics and pathology The patient in the reported case had a recurrence Disease requiring additional therapy, but was alive without evidence of disease at last follow-up, 28 years after Anaplastic large cell lymphoma, ALK-positive diagnosis. Phenotype/cell stem origin Mature (peripheral) T cell. Cytogenetics Etiology Note No etiologic factors are known. Deep RNA sequencing of tumor tissue identified a Epidemiology chimeric transcript fusing the end of exon 6 of The single reported case occurred in an adult male TRAF1 to the start of exon 20 of ALK. (Feldman et al., 2013). The TRAF1-ALK fusion transcript was confirmed at the mRNA level by Sanger sequencing and the Clinics encoded fusion protein was visualized by Western Presentation in the single reported case was with blot. lymphadenopathy and rash. Cytogenetics morphological Pathology Karyotypic findings have not been reported. The pathologic findings in the single reported case were typical for the so-called "lymphohistiocytic Additional anomalies pattern" previously reported in ALK-positive Unknown. ALCLs. Treatment Variants The patient in the reported case was treated with Unknown.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 145 t(2;9)(p23;q33) TRAF1/ALK Xing X, Feldman AL

ALCL, ALK-positive, with t(2;9)(p23;q33) TRAF1/ALK. H&E stain of paraffin embedded tumor tissue shows large atypical cells with cytologic features of "hallmark" cells characteristic of ALCL. Immunohistochemical staining for CD30 shows strong positivity in the tumor cells, with a membranous and Golgi zone distribution. Staining for ALK shows strong cytoplasmic positivity without nuclear staining. The absence of nuclear staining is characteristic for an alternate (non-NPM1) ALK fusion partner. TRAF1 was identified as the partner gene by RNA sequencing.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 146 t(2;9)(p23;q33) TRAF1/ALK Xing X, Feldman AL

Genes involved and therapeutic target. proteins Result of the chromosomal TRAF1 anomaly Location Hybrid gene 9q33.2 Description Protein Expressed, as demonstrated by next-generation and TRAF1 encodes the TRAF1 protein, a member of Sanger sequencing. the tumor necrosis factor receptor-associated factor Fusion protein family of signaling proteins. TRAF1 associates with, and mediates signal transduction from, Note various receptors of the TNFR superfamily. TRAF1 The TRAF1-ALK fusion transcript and TRAF1- and TRAF2 form a heterodimeric complex, which ALK fusion protein both were expressed in the is required for TNF-alpha-mediated activation of reported case. MAPK8/JNK and NF-kappaB. The function of the fusion has not been reported. ALK Description Expressed, as demonstrated by Western blot and Location immunohistochemistry. 2p23.2 Protein References ALK encodes a receptor tyrosine kinase, the Feldman AL, Vasmatzis G, Asmann YW, Davila J, Middha anaplastic lymphoma kinase (ALK), which belongs S, Eckloff BW, Johnson SH, Porcher JC, Ansell SM, to the insulin receptor superfamily and is critical in Caride A. Novel TRAF1-ALK fusion identified by deep RNA the development of the brain. ALK fusion proteins sequencing of anaplastic large cell lymphoma. Genes are critical in the pathogenesis of ALK-positive Chromosomes Cancer. 2013 Nov;52(11):1097-102 ALCLs and a variety of other hematopoietic and This article should be referenced as such: non-hematopoietic neoplasms, in which they serve Xing X, Feldman AL. t(2;9)(p23;q33) TRAF1/ALK. Atlas both as a diagnostic biomarker and potential Genet Cytogenet Oncol Haematol. 2015; 19(2):145-147.

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Leukaemia Section Short Communication t(9;15)(p13;q24) PAX5/GOLGA6A Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)

Published in Atlas Database: June 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0915p13q24ID1596.html DOI: 10.4267/2042/56302 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Protein 391 amino acids; from N-term to C-term, PAX5 Review on t(9;15)(p13;q24) PAX5/GOLGA6A, contains: a paired domain (aa: 16-142); an with data on clinics, and the genes implicated. octapeptide (aa: 179-186); a partial homeodomain (aa: 228-254); a transactivation domain (aa: 304- Clinics and pathology 359); and an inhibitory domain (aa: 359-391). Lineage-specific transcription factor; recognizes the Disease concensus recognition sequence B-cell acute lymphoblastic leukemia (B-ALL) GNCCANTGAAGCGTGAC, where N is any Note nucleotide. Involved in B-cell differentiation. Entry Was not taken into account in this review a case of of common lymphoid progenitors into the B cell diffuse large B-cell lymphoma in a 51 year-old lineage depends on E2A, EBF1, and PAX5; male patient (Morrison et al., 1994). activates B-cell specific genes and repress genes involved in other lineage commitments. Epidemiology Activates the surface cell receptor CD19 and One case to date, a 21-year-old male patient with a repress FLT3. CD10+ (B-II, common) ALL (Coyaud et al., 2010). Pax5 physically interacts with the RAG1/RAG2 Prognosis complex, and removes the inhibitory signal of the lysine-9-methylated histone H3, and induces V-to- No data. DJ rearrangements. Genes repressed by PAX5 expression in early B Cytogenetics cells are restored in their function in mature B cells Cytogenetics morphological and plasma cells, and PAX5 repressed (Fuxa et al., 2004; Johnson et al., 2004; Zhang et al., 2006; There were additional numerial anomalies (+5, Cobaleda et al., 2007; Medvedovic et al., 2011). +21). GOLGA6A Genes involved and Location proteins 15q24.1 Protein PAX5 693 amino acids (aa); contains a coiled coil domain Location (aa 14-611), with 11 leucine zipper motifs (aa 360- 9p13.2 430).

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PAX5/GOLGA6A fusion protein.

Member of the golgin family of proteins, localizes Busslinger M. Pax5 induces V-to-DJ rearrangements and to the Golgi apparatus. Its function is poorly known locus contraction of the immunoglobulin heavy-chain gene. Genes Dev. 2004 Feb 15;18(4):411-22 (Gilles et al., 2000). Johnson K, Pflugh DL, Yu D, Hesslein DG, Lin KI, Bothwell AL, Thomas-Tikhonenko A, Schatz DG, Calame K. B cell- Result of the chromosomal specific loss of histone 3 lysine 9 methylation in the V(H) locus depends on Pax5. Nat Immunol. 2004 Aug;5(8):853- anomaly 61 Hybrid gene Zhang Z, Espinoza CR, Yu Z, Stephan R, He T, Williams GS, Burrows PD, Hagman J, Feeney AJ, Cooper MD. Description Transcription factor Pax5 (BSAP) transactivates the RAG- Fusion of PAX5 exon 6 to GOLGA6A exon 3. mediated V(H)-to-DJ(H) rearrangement of immunoglobulin genes. Nat Immunol. 2006 Jun;7(6):616-24 Fusion protein Cobaleda C, Schebesta A, Delogu A, Busslinger M. Pax5: Description the guardian of B cell identity and function. Nat Immunol. 887 amino acids. The predicted fusion protein 2007 May;8(5):463-70 contains the DNA binding paired domain, the Coyaud E, Struski S, Prade N, Familiades J, Eichner R, octapeptide, and the homeodomain of PAX5 and Quelen C, Bousquet M, Mugneret F, Talmant P, Pages part of the coiled coil domain of GOLGA6A. MP, Lefebvre C, Penther D, Lippert E, Nadal N, Taviaux S, Poppe B, Luquet I, Baranger L, Eclache V, Radford I, Barin C, Mozziconacci MJ, Lafage-Pochitaloff M, Antoine-Poirel References H, Charrin C, Perot C, Terre C, Brousset P, Dastugue N, Broccardo C. Wide diversity of PAX5 alterations in B-ALL: Morrison VA, Frizzera G, Arthur DC, Ogle KM, Hurd DD, a Groupe Francophone de Cytogenetique Hematologique Bloomfield CD, Peterson BA. Prognostic factors for study. Blood. 2010 Apr 15;115(15):3089-97 therapeutic outcome of diffuse small non-cleaved cell lymphoma in adults. Am J Hematol. 1994 Aug;46(4):295- Medvedovic J, Ebert A, Tagoh H, Busslinger M. Pax5: a 303 master regulator of B cell development and leukemogenesis. Adv Immunol. 2011;111:179-206 Gilles F, Goy A, Remache Y, Manova K, Zelenetz AD. Cloning and characterization of a Golgin-related gene from This article should be referenced as such: the large-scale polymorphism linked to the PML gene. Genomics. 2000 Dec 15;70(3):364-74 Huret JL. t(9;15)(p13;q24) PAX5/GOLGA6A. Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2):148-149. Fuxa M, Skok J, Souabni A, Salvagiotto G, Roldan E,

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Solid Tumour Section Short Communication

Bone: Epithelioid hemangioendothelioma Andreas F Mavrogenis, Andrea Angelini, Costantino Errani, Pietro Ruggieri First Department of Orthopaedics, Athens University Medical School, ATTIKON University Hospital, Athens, Greece (AFM), Istituto Ortopedico Rizzoli, Bologna, Italy (AA, CE, PR)

Published in Atlas Database: June 2014 Online updated version : http://AtlasGeneticsOncology.org/Tumors/EpitHemangioendotBoneID5617.html DOI: 10.4267/2042/56303 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Clinics and pathology Review on epithelioid hemangioendothelioma with Note data on clinics and the genes implicated. Pathogenesis unclear. Highly malignant vascular tumors of bone (angiosarcomas) may arise at sites Identity of prior radiation. Other names Epidemiology Hemangiosarcoma Epithelioid hemangioendothelioma of bone account Hemangioendothelioma for less than 1% of malignant bone tumors. It may Hemangioendothelial sarcoma occur at any age, although approximately half of the Epithelioid angiosarcoma cases tend to occur during the second and third Epithelioid sarcoma-like hemangioendothelioma decades of life. Males and females are Pseudomyogenic hemangioendothelioma approximately equally affected. The tumors show a Note wide skeletal distribution affecting the long tubular Vascular tumors of bone range from benign bones of the extremity and the axial skeleton, hemangioma to highly malignant angiosarcoma. mainly the spine. The lower extremities are They are composed of tumor cells forming vascular predominantly affected, with more than half of the spaces. In 1943, Stout defined the diagnostic lesions located in the tibia or femur; spinal lesions criteria for malignant vascular tumors, which he account for less than 10% of the cases. named hemangioendothelioma. Since 1994, the use Approximately one third are multicentric within a of the term "hemangioendothelioma" referring to bone or multifocal, within multiple bones with vascular tumors of bone has decreased due to the lesions randomly distributed throughout the need of a more accurate classification. skeleton or clustered in an anatomic region, such as According to the WHO Classification of Tumors of a single extremity. However, the distinction Soft Tissue and Bone and the ISSVA classification, between multifocal and metastatic disease is not the term "hemangioendothelioma" connotes clear. In general multifocal disease is thought to be intermediate malignancy, except in the context of limited to a specific anatomic region (i.e., bones of epithelioid hemangioendothelioma, which is the same limb) with variable involvement of described as a distinct entity and classified as individual osseous elements. Thus it is debatable malignant. We consider the name epithelioid whether disease in the femur and cervical spine hemangioendothelioma of bone for low-grade may be considered multifocal, as opposed to malignant endothelial vascular neoplasms of bone metastatic given that this tumor has capacity of with tumor cells showing endothelial metastasizing hematologically. However, when differentiation, and a biologic behavior between disease is located in the distal femur or patella, is that of hemangioma and angiosarcoma. easier to consider it multifocal.

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Figure A: Sagittal T2-weighted magnetic resonance imaging of the sacrum shows osteolysis and destruction of the coccyx with anterior soft tissue mass. Biopsy showed epithelioid hemangioendothelioma of the coccyx. Figure B: Sagittal T1-weighted magnetic resonance imaging of the thoracic spine of a patient with recurrent epithelioid hemangioendothelioma of the T3 vertebra.

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Figure C: Photomicrograph (stain, hematoxylin and eosin; original magnification, 10x) shows epithelioid cytomorphology with variably solid or vasoformative architecture and tumoral cells with large, pleomorphic and mildy hyperchromatic nuclei with evident nucleoli. Vascular cavities are variable and matted with neoplastic endothelial cells. Figure D: Photomicrograph (stain, hematoxylin and eosin; original magnification, 20x) shows blood filled cavities of different caliber rimmed by plump endothelium with epithelioid appearance. CD31 staining of the endothelium of the tumoral vessels is typically cytoplasmic.

Clinics Pathology Clinical symptoms include pain and possible Gross pathology: Macroscopically, epithelioid association with a palpable tumor mass. hemangioendothelioma of bone tends to be firm Neurological symptoms may occur in patients with and tan-white. The tumor can erode the cortex and spinal involvement. Tumor growth may be rapid or extend into the soft tissue. slow, and infiltrative. Micropathology: Microscopically, the tumor is Imaging composed of anastomosing cords, solid nests, and The imaging appearance of epithelioid strands of endothelial cells that may sometimes hemangioendothelioma of bone is non-specific. form narrow vascular channels. The small capillary- The tumors are purely lytic, poorly marginated with sized tumor vessels can mimic small reactive varying degrees of peripheral sclerosis. vessels of granulation tissue. The epithelioid cells A soft tissue mass is often associated with less well tend to have eosinophilic cytoplasm which may differentiated tumors. Clustering of multifocal show vacuolization and sometimes signet ring-like lesions in a single anatomic location suggests the appearance. The connective tissue stroma shows diagnosis of a vascular neoplasm. significant myxoid and hyalinized appearance.

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The nuclei of the neoplastic cells show varying genetic analysis, patients with unifocal tumors that degrees of pleomorphism and anaplasia. developed bone lesions after treatment should Although many variants of hemangioendothelioma probably be considered as tumor progression in have been reported, the striking features of growth multifocal form. of epithelioid hemangioendothelioma of bone are Prognosis the formation of atypical endothelial cells (marked nuclear atypia, mitotic activity, spindling of cells The histological degree of differentiation as and necrosis) arranged in cords, in greater numbers evaluated pathologically by the histological pattern than required to line the vessels with a simple of the tumor and the cytologic atypia of the endothelial membrane, and the formation of neoplastic endothelial cells, and the presence of vascular tubes with a delicate framework of unifocal or multifocal tumor are the most important reticulin fibers with a marked tendency for their prognostic factors. lumens to anastomose. On hematoxylin and eosin The survival advantage for the patients with stains the neoplastic epithelioid endothelial cells are multifocal tumors may in part be related to the fact embedded in a hyalinized (deep pink) or chondroid- that multifocal tumors show better differentation. like (light blue) matrix. No tumor should be Conventionally, multifocal disease in a tumor with considered an epithelioid hemangioendothelioma of atypia would be considered a metastatic deposit, bone unless these criteria are present. whereas when it occurs in tumors with entirely Immunophenotype: The endothelial cells benign features it is considered a multifocal uniformly express vimentin and many cells stain process. Additionally, tumor location in the axial with antibodies to Factor VIII, CD31, CD34, and skeleton and limb girdles probably precludes good Ulex Europaeus. Epithelioid malignancies may also prognosis because it is difficult to obtain adequate express cytokeratins and EMA. surgery, or multifocal tumors with poor outcome Ultrastructure: The endothelial cells contain represent tumors with biologic behavior closer to Weibel-Palade bodies, but are generally difficult to angiosarcoma. find in poorly differentiated tumors. Cytoplasmic filaments are abundant. Genetics Treatment Note Surgical: Patients with epithelioid Two epithelioid hemangioendotheliomas have hemangioendothelioma of bone may be cured by shown an identical chromosomal translocation surgery, with or without other treatments such as involving chromosomes 1 and 3. chemotherapy, radiation therapy and embolization. A possible major role for wide surgery should be Genes involved and considered for these tumors whenever this choice does not involve high morbidity or poor functional proteins results. Note Radiation therapy: Adjuvant radiation therapy is Recent identification of WWTR1-CAMTA1 fusion advocated to decrease the risk of local recurrence. provides a powerful diagnostic tool that can be used The risk for postradiation complications should be to distinguish an epithelioid hemangioendothelioma considered. of bone from a hemangioendothelioma. However, Embolization: As a vascular tumor the potential genetic hallmarks of hemangioendothelioma are for intraoperative blood loss is significant. To still under investigation. lessen this complication, patients should have an angiographic evaluation and selective embolization References when feasible. Mallory FB. THE RESULTS OF THE APPLICATION OF Evolution SPECIAL HISTOLOGICAL METHODS TO THE STUDY Outcome: The reported local recurrence rate is up OF TUMORS. J Exp Med. 1908 Sep 5;10(5):575-93 to 13%. Wide tumor resection has been related with Stout AP. HEMANGIO-ENDOTHELIOMA: A TUMOR OF a lower risk for local recurrence; however, a BLOOD VESSELS FEATURING VASCULAR ENDOTHELIAL CELLS. Ann Surg. 1943 Sep;118(3):445- statistically significantly higher survival to local 64 recurrence has not been shown in multivariate analysis. A median survival of 21 months, a 5-year Unni KK, Ivins JC, Beabout JW, Dahlin DC. Hemangioma, hemangiopericytoma, and hemangioendothelioma survival of approximately 33%, and a metastatic (angiosarcoma) of bone. Cancer. 1971 Jun;27(6):1403-14 rate of up to 31% has been reported. It is difficult to Rosai J, Gold J, Landy R. The histiocytoid hemangiomas. establish correctly which patient had bone A unifying concept embracing several previously described metastasis and which patient had progression of the entities of skin, soft tissue, large vessels, bone, and heart. disease in the multifocal form. In the absence of a Hum Pathol. 1979 Nov;10(6):707-30

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Campanacci M, Boriani S, Giunti A. Hisaoka M, Okamoto S, Aoki T, Yokoyama K, Hashimoto Hemangioendothelioma of bone: a study of 29 cases. H.. Spinal epithelioid hemangioendothelioma with Cancer. 1980 Aug 15;46(4):804-14 epithelioid angiosarcomatous areas. Skeletal Radiol. 2005 Nov;34(11):745-9. Epub 2005 May 5. Wold LE, Unni KK, Beabout JW, Ivins JC, Bruckman JE, Dahlin DC. Hemangioendothelial sarcoma of bone. Am J Gupta A, Saifuddin A, Briggs TW, Flanagan AM.. Surg Pathol. 1982 Jan;6(1):59-70 Subperiosteal hemangioendothelioma of the femur. Skeletal Radiol. 2006 Oct;35(10):793-6. Epub 2006 Jan Tsuneyoshi M, Dorfman HD, Bauer TW. Epithelioid 19. hemangioendothelioma of bone. A clinicopathologic, ultrastructural, and immunohistochemical study. Am J Surg Errani C, Zhang L, Sung YS, Hajdu M, Singer S, Maki RG, Pathol. 1986 Nov;10(11):754-64 Healey JH, Antonescu CR.. A novel WWTR1-CAMTA1 gene fusion is a consistent abnormality in epithelioid Resnick D, Kyriakos M, Greenway GD.. Tumors and hemangioendothelioma of different anatomic sites. Genes tumor-like lesions of the bone: Imaging and pathology of Chromosomes Cancer. 2011 Aug;50(8):644-53. doi: specific lesions. In: Resnick D, editor. Diagnosis of bone 10.1002/gcc.20886. Epub 2011 May 16. and joint disorders, 3rd edition. Philadelphia, PA: WB Saunders Co; 1995;3628-938. Hornick JL, Fletcher CD.. Pseudomyogenic hemangioendothelioma: a distinctive, often multicentric Kleer CG, Unni KK, McLeod RA.. Epithelioid tumor with indolent behavior. Am J Surg Pathol. 2011 hemangioendothelioma of bone. Am J Surg Pathol. 1996 Feb;35(2):190-201. doi: 10.1097/PAS.0b013e3181ff0901. Nov;20(11):1301-11. Errani C, Vanel D, Gambarotti M, Alberghini M, Picci P, Mendlick MR, Nelson M, Pickering D, Johansson SL, Faldini C.. Vascular bone tumors: a proposal of a Seemayer TA, Neff JR, Vergara G, Rosenthal H, Bridge classification based on clinicopathological, radiographic JA.. Translocation t(1;3)(p36.3;q25) is a nonrandom and genetic features. Skeletal Radiol. 2012 aberration in epithelioid hemangioendothelioma. Am J Dec;41(12):1495-507. doi: 10.1007/s00256-012-1510-6. Surg Pathol. 2001 May;25(5):684-7. Epub 2012 Sep 21. (REVIEW) Roessner A, Boehling T.. Angiosarcoma. Pathology and Palmerini E, Maki RG, Staals EL, Alberghini M, Antonescu genetics of tumours of soft tissue and bone. In: World CR, Ferrari C, Ruggieri P, Mavrogenis A, Bertoni F, Cesari Health Organization classification of tumours of soft tissue M, Paioli A, Marchesi E, Picci P, Ferrari S.. Primary and bone. Fletcher CDM, Unni KK, Mertens F, editors. Angiosarcoma of Bone: A Retrospective Analysis of 60 Lyon: IARC Press; 2002;322-3. Patients From 2 Institutions. Am J Clin Oncol. 2013 Mar 4. Evans HL, Raymond AK, Ayala AG.. Vascular tumors of [Epub ahead of print] bone: A study of 17 cases other than ordinary Angelini A, Mavrogenis AF, Gambarotti M, Merlino B, Picci hemangioma, with an evaluation of the relationship of P, Ruggieri P.. Surgical treatment and results of 62 hemangioendothelioma of bone to epithelioid patients with epithelioid hemangioendothelioma of bone. J hemangioma, epithelioid hemangioendothelioma, and Surg Oncol. 2014 Jun;109(8):791-7. doi: high-grade angiosarcoma. Hum Pathol. 2003 10.1002/jso.23587. Epub 2014 Mar 18. Jul;34(7):680-9. Aflatoon K, Staals E, Bertoni F, Bacchini P, Donati D, This article should be referenced as such: Fabbri N, Boriani S, Frassica FJ.. Hemangioendothelioma Mavrogenis AF, Angelini A, Errani C, Ruggieri P. Bone: of the spine. Clin Orthop Relat Res. 2004 Jan;(418):191-7. Epithelioid hemangioendothelioma. Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2):150-154.

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 154

Atlas of Genetics and Cytogenetics

in Oncology and Haematology

OPEN ACCESS JOURNAL INIST -CNRS

Deep Insight Section

Insights into structure and function of human PWWP domains

Su Qin, Jinrong Min Structural Genomics Consortium, University of Toronto, 101 College Street, Toronto, Ontario M5G 1L7, Canada (SQ, JM), Department of Physiology, University of Toronto, Toronto, Ontario M5S 1A8, Canada (JM)

Published in Atlas Database: June 2014 Online updated version : http://AtlasGeneticsOncology.org/Deep/PWWPDomainsID20137.html DOI: 10.4267/2042/56304 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2015 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract The PWWP domain is a small structural unit that can bind both DNA and histone, hence involved in chromatin targeting. Here we summarize the structural features of the human PWWP domains and highlight their ligand binding ability from a structural view. Their association with biological functions is also discussed.

Introduction antiparallel β-strands ( β1-β5). The linker between β2 and β3 may also contain additional secondary The PWWP domain is a structural unit of 100-150 elements. The unique feature of the PWWP domain amino acids and named after the conserved Pro- is the presence of a α-bundle of 1-6 α-helixes Trp-Trp-Pro motif (Stec et al., 1998; Stec et al., following the β-barrel (Wu et al., 2011). This α- 2000). It is found throughout eukaryotes, ranging bundle region is very variable and diverse at both from unicellular organisms to human. There are the sequence and structural levels. Therefore, only more than 20 PWWP domain containing proteins in the β-barrel part of the PWWP domain could be the human genome, most of which are chromatin reliably predicted in protein domain databases, such associated. Moreover, the PWWP domain is often as SMART and Human Protein Reference found to coexist with other chromatin-associated Database. The conserved Pro-Trp-Trp-Pro motif is domains, such as the PHD domain and located at the beginning of the β2 strand and it is Bromodomain, in a single polypeptide. More and packed against the α-bundle, indicating its critical more studies are revealing the involvement of the roles in protein folding and stability. Generally PWWP domain in chromatin biology. Here, we will speaking, the PWWP domain can fold as an first summarize the structural features of the independent functional unit. However, recent PWWP domains, then move to their ligand binding studies reveal that the PWWP domain of ability, and finally discuss their association with ZMYND11 (also known as BS69) functions biological functions. together with the preceding Bromodomain and zinc Structural characteristics finger as an integral functional module (Wang et al., 2014; Wen et al., 2014). The PWWP domain belongs to the Royal superfamily, which also includes chromodomain, DNA binding ability of the PWWP Tudor domain, and MBT domain (Maurer-Stroh et domain al., 2003). The Royal superfamily shares a common structural feature, i.e., an antiparallel β-barrel-like The first three-dimensional structure of a PWWP structure formed by 3-5 β-strands. For PWWP domain was determined for the mouse DNA domains, the β-barrel is consisted with 5 methyltransferase Dnmt3b (Qiu et al., 2002).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 155 Bone: Epithelioid hemangioendothelioma Mavrogenis AF, et al.

Figure 1. Complex structure of BRPF1-PWWP and H3K36me3 (PDB ID: 2X4X). Green: H3K36me3 peptide; magenta: β barrel; cyan: α-bundle; yellow: insertion elements between β2 and β3. Residues of the conserved PWWP motif, aromatic cage and methylated K36 are shown in stick mode.

Structural analysis of this PWWP domain revealed recognize methylated lysine and arginine (Adams- a prominent positively charged surface, suggesting Cioaba and Min, 2009), prompted people to a potential role in DNA binding (Qiu et al., 2002). propose the PWWP domain as a potential histone Moreover, it is a common feature that the PWWP "reader" in 2005 (Nameki et al., 2005). Later on, it domain is rich in lysine and arginine residues and was demonstrated that the zebrafish Brpf1-PWWP has a predicted pI of more than 9. Indeed, other domain can bind histones directly (Laue et al., PWWP domains have been reported to be able to 2008) and the fission yeast protein Pdp1 can bind DNA, such as HDGF (Lukasik et al., 2006; recognize H4K20me specifically (Wang et al., Yang and Everett, 2007), MSH6 (Laguri et al., 2009; Qiu et al., 2012). The crystal structure of 2008), PSIP1 (also known as LEDGF and p75) BRPF1-H3K36me3 complex fully established the (Eidahl et al., 2013; van Nuland et al., 2013), and notion that the PWWP domain can recognize ZMYND11 (Wang et al., 2014). In vitro binding methylated histone (Vezzoli et al., 2010; Wu et al., assays revealed that the PWWP domain binds DNA 2011). After that, many other PWWP domains were in a nonspecific manner (Qiu et al., 2002; Lukasik reported to recognize methylated histone peptides, et al., 2006; Laguri et al., 2008). e.g., DNMT3A-H3K36me3 (Dhayalan et al., 2010), To date, no structure of a PWWP-DNA complex is PSIP1-H3K36me3 (Pradeepa et al., 2012; Eidahl et available in protein structure database. Facilitated al., 2013; van Nuland et al., 2013), MSH6- by NMR chemical shift perturbation experiments, H3K36me3 (Li et al., 2013), HDGF2-H3K79me3 several groups tried to map the DNA binding site (Wu et al., 2011), HDGF2-H4K20me3 (Wu et al., on different PWWP domains (HDGF (Lukasik et 2011), and ZMYND11-H3.3K36me3 (Wen et al., al., 2006), MSH6 (Laguri et al., 2008), and PSIP1 2014). (Eidahl et al., 2013; van Nuland et al., 2013)). Structural analysis of these PWWP-histone Notably, the perturbed residues are consistently complex structures identified a conserved cage localized on one side of the protein, centering on formed by three aromatic residues for methyl-lysine the β1-β2 arch region and the PWWP motif, which binding. The third residue (W/Y) of the PWWP also overlaps with the patch of highly positively motif and the residue (F/Y/W) immediately charged surface area. preceding this motif are involved in forming this Histone binding ability of the cage. Another aromatic residue (F/Y/W) comes from the end of β3 strand (Wu et al., 2011). PWWP domain Sequence alignment reveals that most PWWP The structural similarity of the PWWP domain with domains have this conserved cage for potential other Royal superfamily members, which can methylated-histone binding (Wu et al., 2011).

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 156 Insights into structure and function of human PWWP domains Qin S, Min J

However, several exceptions exist. The PWWP mark, they are involved in diverse biological domains of RBBP1, RBBP1L1, MBD5, and NSD1 processes, depending on the protein context they (N-terminal) have incomplete aromatic cage. settled. RBBP1-PWWP indeed did not show any binding to The first class of PWWP domain containing methylated histone peptides (Gong et al., 2012). proteins is directly involved in modifying The histone variant H3.3 possesses a signature chromatin. DNMT3A/B are the de novo DNA motif 'S31...A87AIG90' that is distinct from the methyltransferases responsible for the 'A31...S87AVM90' sequence signature of the establishment of DNA methylation patterns during canonical histone H3.1/2. development (Chédin, 2011). The interaction of the The Bromo-Zinc-PWWP cassette of ZMYND11 PWWP domain with H3K36me3 is involved in specifically recognizes H3.3K36me3 (Wen et al., targeting of DNMT3A to chromatin and guides 2014). In addition to the K36me3 recognition by DNA methylation (Dhayalan et al., 2010). the conserved aromatic cage of PWWP domain, Disruption of the PWWP domain abolishes the S31 of H3.3 is specifically recognized by the ability of DNMT3A and DNMT3B to methylate the Bromodomain and Zinc finger domains major satellite repeats at pericentric synergistically. Glu251 from the Bromodomain and heterochromatin (Chen et al., 2004; Ge et al., Asn266 from the Zinc finger form hydrogen bonds 2004). NSD1/2/3 are histone mono- and di- with the hydroxyl group of S31. methylases for H3K36 (Wagner and Carpenter, Nucleosome binding ability of the 2012). Histone-modifying enzymes often contain "Reader" domains to bind their product to PWWP domain propagate the resultant mark along the chromatin. The mentioned PWWP-binding sites H3K36, Indeed, the N-terminal PWWP domains of H3K79 and H4K20 are at a close proximity to NSD2/NSD3 are able to recognize H3K36me2/3 in DNA in nucleosome context. The fact that PWWP vitro (Wu et al., 2011). GLYR1 (also known as N- domain can bind both DNA and methylated histone PAC), was found to be a cofactor of H3K4 suggests a synergy mechanism among these demethylase LSD2, and GLYR1 positively interactions. The binding affinities of PWWP regulates the H3K4 demethylase activity (Fang et domain towards histone peptide and DNA oligos al., 2013). BRPF1/2/3 are scaffold proteins of the are relatively too weak. But the affinity of PSIP1- MOZ/MORF histone acetyltransferase complexes PWWP for methylated nucleosomes ( Kd ~1.5 mM) (Ullah et al., 2008). Brpf1 is required for histone is four orders of magnitude higher than for a acetylation, the maintenance of cranial Hox gene methylated peptide ( Kd ~17 mM) and two orders expression, and the proper determination of higher than for isolated DNA ( Kd ~150 mM) (van pharyngeal segmental identities during Nuland et al., 2013). In another study, similar trend development of zebrafish (Laue et al., 2008) and was observed for PSIP1: H3KC36me3 nucleosome medaka fish (Hibiya et al., 2009). PWWP domains (Kd ~48 nM), H3K36me3 peptide ( Kd >6.5 mM), of BRPF1/2 are able to recognize H3K36me3 and DNA ( Kd ~1.5 mM) (Eidahl et al., 2013). (Vezzoli et al., 2010; Wu et al., 2011), a mark Studies on ZMYND11 (Wen et al., 2014) and present in transcribed Hox genes (Wang et al., fission yeast Pdp1 (Qiu et al., 2012) show that a 2007), consistent with BRPF1's crucial role in synergy was not detected in vitro assays using promoting the expression of these genes. histone peptides and DNA oligos, so it is likely that The second class of PWWP domain containing the synergy between histone and DNA may proteins is involved in DNA repair. MSH6 binds to contribute only at the nucleosomal level. H3K36me3 mark in a PWWP-dependent manner Strikingly, in all the available histone-PWWP and this interaction mediates MutSa (one of the two complex structures, the histone peptides bind to DNA-mispair recognizing complexes) association PWWP domains in a similar orientation, that is, the with chromatin in cells (Li et al., 2013), hence peptides is almost perpendicular to the β4 strand. contributing to DNA mismatch repair. MUM1 (also Further comparing the DNA binding surface known as EXPAND1) contributes to the chromatin identified by NMR and the histone binding mode architecture and, via its direct interaction with the suggests that PWWP domains may bind to DNA damage mediator 53BP1, plays an accessory nucleosome through a conserved mode, i.e., the role to facilitate damage-induced chromatin PWWP domains adopt distinct and conserved changes and is important for efficient DNA repair interfaces to engage histone and DNA, respectively. and cell survival following DNA damage (Huen et al., 2010). In vivo chromatin association of MUM1 Functional roles of PWWP relies on its PWWP domain-mediated binding to domain containing proteins nucleosomes, and in vitro assays revealed its Although the PWWP domains mainly function as preference of H3K36me3 (Wu et al., 2011). PSIP1 chromatin interactors and recognize H3K36me3 promotes the repair of double strand breaks (DSBs)

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 157 Bone: Epithelioid hemangioendothelioma Mavrogenis AF, et al.

by the homologous recombination repair pathway PWWP domains are engaged in many essential (Daugaard et al., 2012). PSIP1 is constitutively cellular processes. associated with chromatin through its PWWP domain that binds preferentially to methyl-lysine References histone markers characteristic of active Hateboer G, Gennissen A, Ramos YF, Kerkhoven RM, transcription units (probably H3K36me3). It also Sonntag-Buck V, Stunnenberg HG, Bernards R. BS69, a binds CtIP in a DNA damage-dependent manner, novel adenovirus E1A-associated protein that inhibits E1A thereby enhancing its tethering to the active transactivation. EMBO J. 1995 Jul 3;14(13):3159-69 chromatin and facilitating its access to DNA DSBs. Ge H, Si Y, Roeder RG. Isolation of cDNAs encoding novel ZMYND11 was originally identified as an transcription coactivators p52 and p75 reveals an alternate adenovirus E1A-binding protein that inhibits the regulatory mechanism of transcriptional activation. EMBO transactivation function of E1A (Hateboer et al., J. 1998a Nov 16;17(22):6723-9 1995). It also functions as a co-repressor of cellular Ge H, Si Y, Wolffe AP. A novel transcriptional coactivator, transcription factors including the c-Myb p52, functionally interacts with the essential splicing factor (Ladendorff et al., 2001) and interacts with a set of ASF/SF2. Mol Cell. 1998b Dec;2(6):751-9 chromatin remodeling factors, including ATP- Stec I, Wright TJ, van Ommen GJ, de Boer PA, van dependent helicases, histone deacetylases, and Haeringen A, Moorman AF, Altherr MR, den Dunnen JT. WHSC1, a 90 kb SET domain-containing gene, expressed histone methyltransferases (Velasco et al., 2006). in early development and homologous to a Drosophila Most recent study reveals that its Bromo-Zinc dysmorphy gene maps in the Wolf-Hirschhorn syndrome finger-PWWP cassette specific recognizes the critical region and is fused to IgH in t(4;14) multiple histone variant H3.3K36me3 and regulates myeloma. Hum Mol Genet. 1998 Jul;7(7):1071-82 transcription elongation (Venkatesh and Workman, Stec I, Nagl SB, van Ommen GJ, den Dunnen JT. The 2014; Wen et al., 2014). Although ZMYND11 is PWWP domain: a potential protein-protein interaction domain in nuclear proteins influencing differentiation? associated with highly expressed genes, it functions FEBS Lett. 2000 May 4;473(1):1-5 as an unconventional transcription co-repressor by modulating RNA polymerase II at the elongation Ladendorff NE, Wu S, Lipsick JS. BS69, an adenovirus E1A-associated protein, inhibits the transcriptional activity stage. Overexpression of ZMYND11 suppresses of c-Myb. Oncogene. 2001 Jan 4;20(1):125-32 cancer cell growth in vitro and tumour formation in mice (Wen et al., 2014). Qiu C, Sawada K, Zhang X, Cheng X. The PWWP domain of mammalian DNA methyltransferase Dnmt3b defines a PSIP1 was first isolated as an transcriptional co- new family of DNA-binding folds. Nat Struct Biol. 2002 activator (Ge et al., 1998a) and it associates with Mar;9(3):217-24 transcriptional activators and components of the Maurer-Stroh S, Dickens NJ, Hughes-Davies L, basal transcriptional machinery including RNA pol Kouzarides T, Eisenhaber F, Ponting CP. The Tudor II subunits (Ge et al., 1998b). It is also an essential domain 'Royal Family': Tudor, plant Agenet, Chromo, subunit of the MLL complex in MLL oncogenic PWWP and MBT domains. Trends Biochem Sci. 2003 transformations via HOX gene regulation Feb;28(2):69-74 (Yokoyama and Cleary, 2008). PSIP1 short isoform Chen T, Tsujimoto N, Li E. The PWWP domain of Dnmt3a (p52) can interact with splicing factors and and Dnmt3b is required for directing DNA methylation to the major satellite repeats at pericentric heterochromatin. contributes to the regulation of alternative splicing. Mol Cell Biol. 2004 Oct;24(20):9048-58 PSIP1 can also interacts with the HIV integrase and directs viral cDNA integration into transcribed Ge YZ, Pu MT, Gowher H, Wu HP, Ding JP, Jeltsch A, Xu GL. Chromatin targeting of de novo DNA genes. Although there are multiple chromatin methyltransferases by the PWWP domain. J Biol Chem. (DNA) binding domains in PSIP1 protein, the 2004 Jun 11;279(24):25447-54 PWWP domain is essential for its in vivo functions, Nameki N, Tochio N, Koshiba S, Inoue M, Yabuki T, Aoki especially due to its recognition of H3K36me3. M, Seki E, Matsuda T, Fujikura Y, Saito M, Ikari M, Watanabe M, Terada T, Shirouzu M, Yoshida M, Hirota H, Perspective Tanaka A, Hayashizaki Y, Güntert P, Kigawa T, Yokoyama It is becoming clear that the PWWP domain S. Solution structure of the PWWP domain of the hepatoma-derived growth factor family. Protein Sci. 2005 functions through its interactions with chromatin Mar;14(3):756-64 with methylated histone mark. However, the structural basis for these interactions in a Lukasik SM, Cierpicki T, Borloz M, Grembecka J, Everett A, Bushweller JH. High resolution structure of the HDGF nucleosome context remains unclear. Given the PWWP domain: a potential DNA binding domain. Protein PWWP domains are often coexist with other Sci. 2006 Feb;15(2):314-23 chromatin interactors, further studies on their Velasco G, Grkovic S, Ansieau S. New insights into BS69 crosstalk are required to understand their biological functions. J Biol Chem. 2006 Jun 16;281(24):16546-50 functions. It is also valuable to consider these Wang GG, Cai L, Pasillas MP, Kamps MP. NUP98-NSD1 interactions in context of the protein/complex they links H3K36 methylation to Hox-A gene activation and settled. Overall, through binding chromatin, the leukaemogenesis. Nat Cell Biol. 2007 Jul;9(7):804-12

Atlas Genet Cytogenet Oncol Haematol. 2015; 19(2) 158 Insights into structure and function of human PWWP domains Qin S, Min J

Yang J, Everett AD. Hepatoma-derived growth factor binds promotes DNA-end resection and homologous DNA through the N-terminal PWWP domain. BMC Mol recombination. Nat Struct Mol Biol. 2012 Aug;19(8):803-10 Biol. 2007 Oct 31;8:101 Gong W, Zhou T, Mo J, Perrett S, Wang J, Feng Y. Laguri C, Duband-Goulet I, Friedrich N, Axt M, Belin P, Structural insight into recognition of methylated histone Callebaut I, Gilquin B, Zinn-Justin S, Couprie J. Human tails by retinoblastoma-binding protein 1. J Biol Chem. mismatch repair protein MSH6 contains a PWWP domain 2012 Mar 9;287(11):8531-40 that targets double stranded DNA. Biochemistry. 2008 Jun 10;47(23):6199-207 Pradeepa MM, Sutherland HG, Ule J, Grimes GR, Bickmore WA. Psip1/Ledgf p52 binds methylated histone Laue K, Daujat S, Crump JG, Plaster N, Roehl HH, Kimmel H3K36 and splicing factors and contributes to the CB, Schneider R, Hammerschmidt M. The multidomain regulation of alternative splicing. PLoS Genet. protein Brpf1 binds histones and is required for Hox gene 2012;8(5):e1002717 expression and segmental identity. Development. 2008 Jun;135(11):1935-46 Qiu Y, Zhang W, Zhao C, Wang Y, Wang W, Zhang J, Zhang Z, Li G, Shi Y, Tu X, Wu J. Solution structure of the Ullah M, Pelletier N, Xiao L, Zhao SP, Wang K, Degerny C, Pdp1 PWWP domain reveals its unique binding sites for Tahmasebi S, Cayrou C, Doyon Y, Goh SL, Champagne methylated H4K20 and DNA. Biochem J. 2012 Mar N, Côté J, Yang XJ. Molecular architecture of quartet 15;442(3):527-38 MOZ/MORF histone acetyltransferase complexes. Mol Cell Biol. 2008 Nov;28(22):6828-43 Wagner EJ, Carpenter PB. Understanding the language of Lys36 methylation at histone H3. Nat Rev Mol Cell Biol. Yokoyama A, Cleary ML. Menin critically links MLL 2012 Jan 23;13(2):115-26 proteins with LEDGF on cancer-associated target genes. Cancer Cell. 2008 Jul 8;14(1):36-46 Eidahl JO, Crowe BL, North JA, McKee CJ, Shkriabai N, Feng L, Plumb M, Graham RL, Gorelick RJ, Hess S, Adams-Cioaba MA, Min J. Structure and function of Poirier MG, Foster MP, Kvaratskhelia M. Structural basis histone methylation binding proteins. Biochem Cell Biol. for high-affinity binding of LEDGF PWWP to 2009 Feb;87(1):93-105 mononucleosomes. Nucleic Acids Res. 2013 Apr 1;41(6):3924-36 Hibiya K, Katsumoto T, Kondo T, Kitabayashi I, Kudo A. Brpf1, a subunit of the MOZ histone acetyl transferase Fang R, Chen F, Dong Z, Hu D, Barbera AJ, Clark EA, complex, maintains expression of anterior and posterior Fang J, Yang Y, Mei P, Rutenberg M, Li Z, Zhang Y, Xu Y, Hox genes for proper patterning of craniofacial and caudal Yang H, Wang P, Simon MD, Zhou Q, Li J, Marynick MP, skeletons. Dev Biol. 2009 May 15;329(2):176-90 Li X, Lu H, Kaiser UB, Kingston RE, Xu Y, Shi YG. LSD2/KDM1B and its cofactor NPAC/GLYR1 endow a Wang Y, Reddy B, Thompson J, Wang H, Noma K, Yates structural and molecular model for regulation of H3K4 JR 3rd, Jia S. Regulation of Set9-mediated H4K20 demethylation. Mol Cell. 2013 Feb 7;49(3):558-70 methylation by a PWWP domain protein. Mol Cell. 2009 Feb 27;33(4):428-37 Li F, Mao G, Tong D, Huang J, Gu L, Yang W, Li GM. The histone mark H3K36me3 regulates human DNA mismatch Dhayalan A, Rajavelu A, Rathert P, Tamas R, Jurkowska repair through its interaction with MutS α. Cell. 2013 Apr RZ, Ragozin S, Jeltsch A. The Dnmt3a PWWP domain 25;153(3):590-600 reads histone 3 lysine 36 trimethylation and guides DNA methylation. J Biol Chem. 2010 Aug 20;285(34):26114-20 van Nuland R, van Schaik FM, Simonis M, van Heesch S, Cuppen E, Boelens R, Timmers HM, van Ingen H. Huen MS, Huang J, Leung JW, Sy SM, Leung KM, Ching Nucleosomal DNA binding drives the recognition of YP, Tsao SW, Chen J. Regulation of chromatin H3K36-methylated nucleosomes by the PSIP1-PWWP architecture by the PWWP domain-containing DNA domain. Epigenetics Chromatin. 2013 May 8;6(1):12 damage-responsive factor EXPAND1/MUM1. Mol Cell. 2010 Mar 26;37(6):854-64 Venkatesh S, Workman JL. Recognizing methylated histone variant H3.3 to prevent tumors. Cell Res. 2014 Vezzoli A, Bonadies N, Allen MD, Freund SM, Santiveri Jun;24(6):649-50 CM, Kvinlaug BT, Huntly BJ, Göttgens B, Bycroft M. Molecular basis of histone H3K36me3 recognition by the Wang J, Qin S, Li F, Li S, Zhang W, Peng J, Zhang Z, PWWP domain of Brpf1. Nat Struct Mol Biol. 2010 Gong Q, Wu J, Shi Y. Crystal structure of human BS69 May;17(5):617-9 Bromo-ZnF-PWWP reveals its role in H3K36me3 nucleosome binding. Cell Res. 2014 Jul;24(7):890-3 Chédin F. The DNMT3 family of mammalian de novo DNA methyltransferases. Prog Mol Biol Transl Sci. Wen H, Li Y, Xi Y, Jiang S, Stratton S, Peng D, Tanaka K, 2011;101:255-85 Ren Y, Xia Z, Wu J, Li B, Barton MC, Li W, Li H, Shi X. ZMYND11 links histone H3.3K36me3 to transcription Wu H, Zeng H, Lam R, Tempel W, Amaya MF, Xu C, elongation and tumour suppression. Nature. 2014 Apr Dombrovski L, Qiu W, Wang Y, Min J. Structural and 10;508(7495):263-8 histone binding ability characterizations of human PWWP domains. PLoS One. 2011;6(6):e18919 This article should be referenced as such: Daugaard M, Baude A, Fugger K, Povlsen LK, Beck H, Qin S, Min J. Insights into structure and function of human Sørensen CS, Petersen NH, Sorensen PH, Lukas C, PWWP domains. Atlas Genet Cytogenet Oncol Haematol. Bartek J, Lukas J, Rohde M, Jäättelä M. LEDGF (p75) 2015; 19(2):155-159.

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