DOCSLIB.ORG

ETV6 Is the Target of Chromosome 12P Deletions in T(12;21) Childhood Acute Lymphocytic Leukemia

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
ETV6 Is the Target of Chromosome 12P Deletions in T(12;21) Childhood Acute Lymphocytic Leukemia Leukemia (1997) 11, 1459–1464 1997 Stockton Press All rights reserved 0887-6924/97 $12.00 ETV6 is the target of chromosome 12p deletions in t(12;21) childhood acute lymphocytic leukemia H Cave´ 1, V Cacheux2, S Raynaud3, G Brunie1, M Bakkus4, P Cochaux5, C Preudhomme6, JL Laı¨7, E Vilmer8 and B Grandchamp1,9 1Laboratoire de Biochimie Ge´ne´tique, 2Laboratoire de Biologie du De´veloppement, 8Service d’He´matologie, Hoˆpital Robert Debre´, Paris; 3Laboratoire de Ge´ne´tique Mole´culaire des Cancers Humains, CNRS URA 1462, Nice, France; 4Faculte´ de Me´decine VUB, Bruxelles; 5Universite´ libre de Bruxelles, De´partement de Ge´ne´tique Me´dicale, Hopital Erasme, Bruxelles, Belgium; 6Institut de recherche sur le cancer, Lille; 7Laboratoire de cytoge´ne´tique, CHRU, Lille; and 9INSERM U409, Faculte´ de Me´decine Xavier Bichat, Paris, France The presence of ETV6 deletions was investigated in 215 chil- of t(5;12)10 and was subsequently found to be fused to a dren with acute lymphoblastic leukemia (ALL) using the loss number of different partners as a result of various leukemia- of heterozygosity (LOH) approach. We used four intragenic or 11–13 juxtagenic microsatellite markers to detect allelic deletions. In associated translocations. In pediatric B lineage ALL, this series of unselected patients, LOH of ETV6 markers was t(12;21)(p13;q22), which leads to the formation of a ETV6- found in 23% of cases (6% of T-ALL and 26% of B lineage ALL) AML1 fusion gene on the der(21) chromosome,14,15 is found confirming that chromosome 12p12-13 deletions represent a with a high frequency (16–28%).16–18 Interestingly, we have major genetic alteration in childhood ALL, frequently missed by previously shown that t(12;21) and deletions of the non-trans- cytogenetic analysis. The presence of a t(12;21)(p13;q22) was located chromosome 12 were frequently associated in these studied by RT-PCR and/or FISH in a total of 134 patients (125 B lineage ALL, nine T-ALL) including 42 cases with LOH. Thirty- children and we found an intragenic deletion of ETV6 in one four out of 44 patients (77%) for whom a t(12;21) was observed patient with a t(12;21) suggesting that ETV6 was the target of displayed LOH of the ETV6 markers. When associated with a the deletions. In our initial series, 14 out of 16 patients with t(12;21), ETV6 is very likely to be the target of deletions as indi- t(12;21) had lost the non-translocated allele of ETV6.6 These cated by the detection of intragenic deletions in three patients. results contrast with those of other studies using Southern blot Although deletion of ETV6 and t(12;21) were associated in most analysis that reported a much lower frequency of ETV6 patients, in eight cases (six B lineage and two T-ALL) LOH was 16,17 detected at the ETV6 locus without ETV6-AML1 hybrid RNA. deletions in patients with a t(12;21). Besides, while FISH studies conducted in five of these eight patients con- t(12;21) is specifically found in children with B lineage ALL, firmed the absence of translocation involving ETV6. In such 12p deletions have also been detected in patients with other patients, the other allele of ETV6 could be disrupted by either types of malignancies19 including T-ALL,20 and it is unclear a small deletion, a point mutation, or an epigenetic modification whether or not ETV6 is the target of 12p deletions in all cases and it will be of interest to study the structure and expression or in a specific subset of patients. Indeed, a recent study of of the remaining allele of ETV6 in these cases. Alternatively, a tumor suppressor gene located close to ETV6 and CDKN1B LOH for 12p markers in childhood ALL suggested that ETV6 could be the target of deletions. might not be the target of all 12p deletions, since a second Keywords: ETV6; ETV6-AML1; t(12;21); loss of heterozygosity; region of deletion that did not include ETV6 was found near deletion; acute lymphoblastic leukemia CDKN1B.4 The aims of the present study were to establish the frequency of ETV6 deletions in a large series of children with B and T lineage ALL, to determine the occurence of small Introduction deletions involving specifically ETV6 that may incriminate this gene as the target gene of the deletions, and to correlate the Abnormalities of the short arm of chromosome 12 are quite presence of these deletions with the presence of t(12;21). We common in various hematological malignancies including used four intragenic or juxtagenic microsatellite markers to acute myeloid leukemia, myelodysplastic syndrome, myelo- detect allelic deletions of ETV6, and the t(12;21) translocation proliferative disease and acute lymphoblastic leukemia (ALL).1 was sought by FISH and/or RT-PCR of the ETV6-AML1 Chromosome 12p alterations consist of both deletions and chimeric transcript. translocations. Recent studies using microsatellite markers have shown that loss of heterozygosity (LOH) in the chromo- somal region 12p12-13 is found in 20–40% of cases in child- Patients and methods hood ALL.2–4 This suggested that a tumor suppressor gene (TSG) might be located in this region whose inactivation Patients would participate in leukemogenesis.5 LOH mapping and FISH analysis indicated that, in the majority of patients, the We studied 215 children (aged 3 months to 15 years) with deletion contained two genes, the CDKN1B gene (previously ALL (183 B lineage ALL, 32 T-ALL). Eight of these children named KIP1) and the ETS-variant gene 6 (ETV6, previously were studied both at diagnosis and at marrow relapse. Chil- named TEL).3,4,6–9 CDKN1B encodes the cyclin-dependent dren were consecutively diagnosed and unselected, except kinase inhibitor p27 that is a regulator of the cell cycle and that inclusion required the presence of at least 70% of blasts therefore a candidate TSG, but several studies concluded that in marrow at the time of diagnosis or relapse (13 children the remaining allele of this gene is not mutated in patients have been excluded because of a low percentage of blasts in with one deleted allele.3,4 ETV6 was cloned at the breakpoint marrow). ALL were classified on the basis of lymphoid mor- phologic appearance and immunophenotype as assessed by flow cytometry. ALL were defined as belonging to the T lin- Correspondence: H Cave´, Laboratoire de Biochimie Ge´ne´tique, Hoˆpi- eage based on the presence of at least two of the T cell anti- tal Robert Debre´, 48 boulevard Se´rurier, 75019 Paris, France gens CD2, CD5 and CD7. The diagnosis of B precursor ALL Received 20 March 1997; accepted 17 June 1997 was based on the expression of B cell-associated antigens ETV6 deletions in childhood B lineage ALL H Cave´ et al 1460 (CD19+, CD22±, CD10±) and the lack of surface membrane lows: 10 s denaturation at 94°C, 10 s annealing extension at immunoglobulins (Ig). Informed consent was obtained from 68°C. Amplification was controlled by electrophoresis in 2% the patients, their parents, or both, as appropriate. agarose and ethidium bromide staining. ETV6-AML1 and AML1-ETV6 PCR gave rise to a 103 bp and a 199 bp fragment, respectively. The quality of the cDNAs was checked by ampli- Preparation of mononuclear cell lysates fying a 155 bp sequence of the porphobilinogene deaminase cDNA using primers P-PBGD (5′-AGCGGAGCCATG- Bone marrow (BM) was collected on EDTA before induction TCTGGTAACGGCAATG-3′)and N-PBGD (5′-AGGCCAGGG- therapy and during clinical remission between 6 and 24 TACGAGGCTTTCAATGTTG-3′) under the same PCR con- months of therapy according to the EORTC 58 881 follow- ditions as described above but with an annealing extension up protocol. Mononuclear cells were separated by gradient temperature of 60°C. centrifugation over Ficoll (Lymphoprep; Pharmacia, Uppsala, Specificity of the amplification was tested by Southern blot Sweden), washed in 0.9% NaCl, counted, and aliquots of cells hybridization to radio-labeled junctional probes for ETV6- were stored at −80°C. Erythrocytes were lysed by resus- AML1 (5′-AGAATAGCAGAATGCATACT-3′).15 pending the cells in a solution containing 10 mm Tris-HCl (pH 7.5), 1% Triton-100 (Sigma, St Louis, MO, USA), 5 mm MgCl2 and 0.32 mm saccharose. After centrifugation, mono- Results nuclear cells were lysed in a buffer containing 0.45% Tween 20 (Difco, Detroit, MI, USA), 1.45% Nonidet P40 (Sigma), Frequency of monoallelic ETV6 deletion in childhood 2.5 mm MgCl2 and 10 mm Tris-HCl (pH 7.5) and incubated ALL for 2 h at 60°C in the presence of 1 mg/ml proteinase K. Vol- umes were adjusted to obtain 6 ng of DNA per ml of lysate. The presence of allelic deletions of ETV6 was investigated in After a 10-min incubation at 95°C, the lysates were stored at 215 children with ALL using the LOH approach. Partial results −20°C until analysis. concerning 66 of these children (including 16 with LOH) have been previously reported.6 The polymorphic markers used for this screening were D12S1697, D12S89, CA159A7 and Search for loss of heterozygosity D12S98. Markers D12S89 and CA159A7 are located within the ETV6 gene. D12S89 has been previously shown to map Seven simple tandem repeat (STR) markers were used that immediately upstream of exon 2 while CA159A7 maps in the span the chromosome bands 12p12-13. These markers are 5′ region of exon 1B.21 D12S1697 is located 7 kb upstream ordered from telomere to centromere: D12S1697, D12S89, of exon 1A of ETV6, and D12S98 is about 50 kb downstream CA159A7, D12S98, D12S1581, D12S1580 and D12S320.3,21 of exon 8.21 Loss of heterozygosity was evidenced by comparison of Percentages of LOH observed for each marker are reported allelic patterns of amplified DNA on the diagnosis sample in Table 1.
Load more

Recommended publications
  • Ubiquitin-Mediated Control of ETS Transcription Factors: Roles in Cancer and Development
    International Journal of Molecular Sciences Review Ubiquitin-Mediated Control of ETS Transcription Factors: Roles in Cancer and Development Charles Ducker * and Peter E. Shaw * Queen’s Medical Centre, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK * Correspondence: [email protected] (C.D.); [email protected] (P.E.S.) Abstract: Genome expansion, whole genome and gene duplication events during metazoan evolution produced an extensive family of ETS genes whose members express transcription factors with a conserved winged helix-turn-helix DNA-binding domain. Unravelling their biological roles has proved challenging with functional redundancy manifest in overlapping expression patterns, a common consensus DNA-binding motif and responsiveness to mitogen-activated protein kinase signalling. Key determinants of the cellular repertoire of ETS proteins are their stability and turnover, controlled largely by the actions of selective E3 ubiquitin ligases and deubiquitinases. Here we discuss the known relationships between ETS proteins and enzymes that determine their ubiquitin status, their integration with other developmental signal transduction pathways and how suppression of ETS protein ubiquitination contributes to the malignant cell phenotype in multiple cancers. Keywords: E3 ligase complex; deubiquitinase; gene fusions; mitogens; phosphorylation; DNA damage 1. Introduction Citation: Ducker, C.; Shaw, P.E. Cell growth, proliferation and differentiation are complex, concerted processes that Ubiquitin-Mediated Control of ETS Transcription Factors: Roles in Cancer rely on careful regulation of gene expression. Control over gene expression is maintained and Development. Int. J. Mol. Sci. through signalling pathways that respond to external cellular stimuli, such as growth 2021, 22, 5119. https://doi.org/ factors, cytokines and chemokines, that invoke expression profiles commensurate with 10.3390/ijms22105119 diverse cellular outcomes.
    [Show full text]
  • Hsa-Mir-125B-2 Is Highly Expressed in Childhood ETV6&Sol;RUNX1
    Leukemia (2010) 24, 89–96 & 2010 Macmillan Publishers Limited All rights reserved 0887-6924/10 $32.00 www.nature.com/leu ORIGINAL ARTICLE Hsa-mir-125b-2 is highly expressed in childhood ETV6/RUNX1 (TEL/AML1) leukemias and confers survival advantage to growth inhibitory signals independent of p53 N Gefen1,2,8, V Binder1,3,8, M Zaliova4, Y Linka3, M Morrow6, A Novosel3, L Edry5, L Hertzberg1,2,7, N Shomron5, O Williams6, J Trka4, A Borkhardt3 and S Izraeli1,2 1Section of Functional Genomics and Leukemia Research, Department of Pediatric Hemato-Oncology, Sheba Medical Center, Tel-Hashomer, Israel; 2Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel; 3Department of Pediatric Hematology, Oncology and Immunology, Heinrich Heine University, Duesseldorf, Germany; 4Department of Pediatric Hematology and Oncology, Second Faculty of Medicine, Charles University and University Hospital Motol, Childhood Leukemia Investigation Prague, Prague, Czech Republic; 5Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel; 6Institute of Child Health, University College London, London, UK and 7Department of Physics of Complex Systems, Weizmann Institute of Science, Rehovot, Israel MicroRNAs (miRNAs) regulate the expression of multiple characterized by an intrachromosomal amplification of a small proteins in a dose-dependent manner. We hypothesized that region within the long arm of chr 21 (iAMP21) around the increased expression of miRNAs encoded on chromosome 21 4 (chr 21) contribute to the leukemogenic function of trisomy 21. RUNX1 locus. In contrast to ETV6/RUNX1 and HHD ALL, it is The levels of chr 21 miRNAs were quantified by qRT–PCR in associated with poor prognosis.
    [Show full text]
  • 12Q Deletions FTNW
    12q deletions rarechromo.org What is a 12q deletion? A deletion from chromosome 12q is a rare genetic condition in which a part of one of the body’s 46 chromosomes is missing. When material is missing from a chromosome, it is called a deletion. What are chromosomes? Chromosomes are the structures in each of the body’s cells that carry genetic information telling the body how to develop and function. They come in pairs, one from each parent, and are numbered 1 to 22 approximately from largest to smallest. Additionally there is a pair of sex chromosomes, two named X in females, and one X and another named Y in males. Each chromosome has a short (p) arm and a long (q) arm. Looking at chromosome 12 Chromosome analysis You can’t see chromosomes with the naked eye, but if you stain and magnify them many hundreds of times under a microscope, you can see that each one has a distinctive pattern of light and dark bands. In the diagram of the long arm of chromosome 12 on page 3 you can see the bands are numbered outwards starting from the point at the top of the diagram where the short and long arms meet (the centromere). Molecular techniques If you magnify chromosome 12 about 850 times, a small piece may be visibly missing. But sometimes the missing piece is so tiny that the chromosome looks normal through a microscope. The missing section can then only be found using more sensitive molecular techniques such as FISH (fluorescence in situ hybridisation, a technique that reveals the chromosomes in fluorescent colour), MLPA (multiplex ligation-dependent probe amplification) and/or array-CGH (microarrays), a technique that shows gains and losses of tiny amounts of DNA throughout all the chromosomes.
    [Show full text]
  • Investigation of Differentially Expressed Genes in Nasopharyngeal Carcinoma by Integrated Bioinformatics Analysis
    916 ONCOLOGY LETTERS 18: 916-926, 2019 Investigation of differentially expressed genes in nasopharyngeal carcinoma by integrated bioinformatics analysis ZhENNING ZOU1*, SIYUAN GAN1*, ShUGUANG LIU2, RUjIA LI1 and jIAN hUANG1 1Department of Pathology, Guangdong Medical University, Zhanjiang, Guangdong 524023; 2Department of Pathology, The Eighth Affiliated hospital of Sun Yat‑sen University, Shenzhen, Guangdong 518033, P.R. China Received October 9, 2018; Accepted April 10, 2019 DOI: 10.3892/ol.2019.10382 Abstract. Nasopharyngeal carcinoma (NPC) is a common topoisomerase 2α and TPX2 microtubule nucleation factor), malignancy of the head and neck. The aim of the present study 8 modules, and 14 TFs were identified. Modules analysis was to conduct an integrated bioinformatics analysis of differ- revealed that cyclin-dependent kinase 1 and exportin 1 were entially expressed genes (DEGs) and to explore the molecular involved in the pathway of Epstein‑Barr virus infection. In mechanisms of NPC. Two profiling datasets, GSE12452 and summary, the hub genes, key modules and TFs identified in GSE34573, were downloaded from the Gene Expression this study may promote our understanding of the pathogenesis Omnibus database and included 44 NPC specimens and of NPC and require further in-depth investigation. 13 normal nasopharyngeal tissues. R software was used to identify the DEGs between NPC and normal nasopharyngeal Introduction tissues. Distributions of DEGs in chromosomes were explored based on the annotation file and the CYTOBAND database Nasopharyngeal carcinoma (NPC) is a common malignancy of DAVID. Gene ontology (GO) and Kyoto Encyclopedia of occurring in the head and neck. It is prevalent in the eastern Genes and Genomes (KEGG) pathway enrichment analysis and southeastern parts of Asia, especially in southern China, were applied.
    [Show full text]
  • Chromosomes 8, 12, and 17 Copy Number in Astler-Coller Stage C Colon Cancer in Relation to Proliferative Activity and DNA Ploidy1
    [CANCER RESEARCH 5.1.681-686, February 1. 1993] Chromosomes 8, 12, and 17 Copy Number in Astler-Coller Stage C Colon Cancer in Relation to Proliferative Activity and DNA Ploidy1 Melissa G. Steiner,2 Seth P. Harlow, Edoardo Colombo,3 and Kenneth D. Bauer4 Department of Pathology and the Cancer Center, Northwestern university, McGaw Medical Center, Chicago. Illinois 60611 ABSTRACT to glass slides; evaluation of the number of signals in each nucleus reflects the number of copies of that chromosome in that nucleus. Fluorescence in situ hybridization using centromere-specific DNA Most ISH studies have been applied to hematological cancers (22-24); probes to chromosomes 8,12, and 17 was applied to 23 archival paraffin- however, a few have addressed the chromosomal anomalies in bladder embedded stage C colonie cancer specimens. Chromosome copy number tumors (4, 25-27) and in primary breast tumors (28-30). Furthermore, was related to flow cytometric determinations of S-phase fraction and techniques of ISH can be applied to archival paraffin-embedded spec DNA ploidy. Three to eight copies of chromosomes 8, 12, and 17 were observed at mean frequencies of 28.7%, 37.8%, and 20.9%, respectively. imens (26, 30). The mean frequency of multiple copies of chromosome 12 was signifi Combining FCM and ISH on the same specimen is generally fea cantly greater than that for chromosome 17 (/' < 0.0025). The mean sible, because only thousands of cells are necessary to analyze the frequency of single copies of chromosome 17 was significantly greater than sample in a statistically évaluablemanner using either method.
    [Show full text]
  • HDAC3: Taking the SMRT-N-Correct Road to Repression
    Oncogene (2007) 26, 5439–5449 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc REVIEW HDAC3: taking the SMRT-N-CoRrect road to repression P Karagianni and J Wong1 Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, USA Known histone deacetylases (HDACs) are divided into repression when targeted to promoters, as well as different classes, and HDAC3 belongs to Class I. Through physical association with the DNA-binding factor forming multiprotein complexes with the corepressors YY1 (Yang et al., 1997; Dangond et al., 1998; Emiliani SMRT and N-CoR, HDAC3 regulates the transcription et al., 1998). Together, these findings suggested that this of a plethora of genes. A growing list of nonhistone ubiquitously expressed protein could be involved in the substrates extends the role of HDAC3 beyond transcrip- regulation of mammalian gene expression. On the other tional repression. Here, we review data on the composi- hand, HDAC3 contains an intriguingly variable C tion, regulation and mechanism of action of the SMRT/ terminus, with no apparent similarity with other N-CoR-HDAC3 complexes and provide several examples HDACs. This observation led to the hypothesis that of nontranscriptional functions, to illustrate the wide HDAC3 may have some unique properties and may variety of physiological processes affected by this deacety- not be completely redundant with the other HDACs lase. Furthermore, we discuss the implication of HDAC3 (Yang et al., 1997). This is also suggested by the in cancer, focusing on leukemia. We conclude with some differential localization of HDAC3, which, unlike the thoughts about the potential therapeutic efficacies of predominantly nuclear HDACs 1 and 2, can be found in HDAC3 activity modulation.
    [Show full text]
  • The Complex Genetic Landscape of Familial MDS and AML Reveals Pathogenic Germline Variants
    ARTICLE https://doi.org/10.1038/s41467-020-14829-5 OPEN The complex genetic landscape of familial MDS and AML reveals pathogenic germline variants Ana Rio-Machin et al.# The inclusion of familial myeloid malignancies as a separate disease entity in the revised WHO classification has renewed efforts to improve the recognition and management of this group of at risk individuals. Here we report a cohort of 86 acute myeloid leukemia (AML) and 1234567890():,; myelodysplastic syndrome (MDS) families with 49 harboring germline variants in 16 pre- viously defined loci (57%). Whole exome sequencing in a further 37 uncharacterized families (43%) allowed us to rationalize 65 new candidate loci, including genes mutated in rare hematological syndromes (ADA, GP6, IL17RA, PRF1 and SEC23B), reported in prior MDS/AML or inherited bone marrow failure series (DNAH9, NAPRT1 and SH2B3) or variants at novel loci (DHX34) that appear specific to inherited forms of myeloid malignancies. Altogether, our series of MDS/AML families offer novel insights into the etiology of myeloid malignancies and provide a framework to prioritize variants for inclusion into routine diagnostics and patient management. #A full list of authors and their affiliations appears at the end of the paper. NATURE COMMUNICATIONS | (2020) 11:1044 | https://doi.org/10.1038/s41467-020-14829-5 | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | https://doi.org/10.1038/s41467-020-14829-5 nherited forms of myeloid malignancies are thought to be rare, Results Ialthough the precise
    [Show full text]
  • An Integrative Approach Reveals Genetic Complexity and Epigenetic
    Human Pathology (2019) 91,1–10 www.elsevier.com/locate/humpath Original contribution An integrative approach reveals genetic complexity and epigenetic perturbation in acute promyelocytic leukemia: a single institution experience☆ Nina Rahimi MD a,YanmingZhangMDb, Alain Mina MD c, Jessica K. Altman MD c, Madina Sukhanova PhD a, Olga Frankfurt MD c, Lawrence Jennings MD, PhD a, Xinyan Lu MD a,AmirBehdadMDa, Qing Chen MD, PhD a, Yi-Hua Chen MD a, Juehua Gao MD, PhD a,⁎ aDepartment of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA bDepartment of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA cDepartment of Hematology and Oncology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA Received 10 March 2019; revised 8 May 2019; accepted 8 May 2019 Keywords: Summary Acute promyelocytic leukemia (APL) is a distinct type of acute myeloid leukemia that is Acute promyelocytic defined by the presence of the translocations that mostly involve the RARA gene. The most frequent leukemia; translocation is the t(15;17), which fuses the RARA gene with the PML gene. Previous studies have Next-generation shown that other cooperative mutations are required for the development of APL after the initiating sequencing; event of the t(15;17). In this study, we combined cytogenetics with next-generation sequencing and Somatic mutations; single-nucleotide polymorphism array to study the genetic complexity in 20 APL cases diagnosed in our in- SNP array; stitution. All but 3 cases had additional genetic aberrations. Our study demonstrated that somatic mutations Copy neutral loss of are frequent events in APL.
    [Show full text]
  • Immunohistochemical Expression of Wilms' Tumor 1 Protein In
    applied sciences Review Immunohistochemical Expression of Wilms’ Tumor 1 Protein in Human Tissues: From Ontogenesis to Neoplastic Tissues Lucia Salvatorelli 1,*, Giovanna Calabrese 2 , Rosalba Parenti 2 , Giada Maria Vecchio 1, Lidia Puzzo 1, Rosario Caltabiano 1 , Giuseppe Musumeci 3 and Gaetano Magro 1 1 Department of Medical and Surgical Sciences and Advanced Technologies, G.F. Ingrassia, Azienda Ospedaliero-Universitaria “Policlinico-Vittorio Emanuele”, Anatomic Pathology Section, School of Medicine, University of Catania, 95123 Catania, Italy; [email protected] (G.M.V.); [email protected] (L.P.); [email protected] (R.C.); [email protected] (G.M.) 2 Department of Biomedical and Biotechnological Sciences, Physiology Section, University of Catania, 95123 Catania, Italy; [email protected] (G.C.); [email protected] (R.P.) 3 Department of Biomedical and Biotechnological Sciences, Human Anatomy and Histology Section, School of Medicine, University of Catania, 95123 Catania, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-095-3702138; Fax: +39-095-3782023 Received: 4 October 2019; Accepted: 10 December 2019; Published: 19 December 2019 Abstract: The human Wilms’ tumor gene (WT1) was originally isolated in a Wilms’ tumor of the kidney as a tumor suppressor gene. Numerous isoforms of WT1, by combination of alternative translational start sites, alternative RNA splicing and RNA editing, have been well documented. During human ontogenesis, according to the antibodies used, anti-C or N-terminus WT1 protein, nuclear expression can be frequently obtained in numerous tissues, including metanephric and mesonephric glomeruli, and mesothelial and sub-mesothelial cells, while cytoplasmic staining is usually found in developing smooth and skeletal cells, myocardium, glial cells, neuroblasts, adrenal cortical cells and the endothelial cells of blood vessels.
    [Show full text]
  • Germline Mutations in Etv6 Result in Autosomal Dominant
    GERMLINE MUTATIONS IN ETV6 RESULT IN AUTOSOMAL DOMINANT THROMBOCYTOPENIA By LEILA J. NOETZLI B.S., University of California San Diego, 2007 A thesis submitted to the Faculty of the Graduate School of the University of Colorado in partial fulfillment of the requirements for the degree of Doctor of Philosophy Human Medical Genetics and Genomics Program 2017 This thesis for the Doctor of Philosophy degree by Leila J. Noetzli has been approved for the Human Medical Genetics and Genomics Program by Robert Sclafani, Chair Jorge Di Paola, Advisor Jay Hesselberth Richard Spritz Rytis Prekeris Date: May 19, 2017 ii Noetzli, Leila J. (Ph.D., Human Medical Genetics and Genomics) Germline mutations in ETV6 result in autosomal dominant thrombocytopenia Thesis directed by Associate Professor Jorge Di Paola ABSTRACT Whole exome sequencing on a family with autosomal dominant thrombocytopenia and two occurrences of acute lymphoblastic leukemia (ALL) of unknown cause identified a heterozygous single nucleotide change in the gene ETV6, encoding a p.Pro214Leu amino acid substitution. We screened families with the same phenotype and found mutations in ETV6 in two additional families: one with the identical p.Pro214Leu mutation and one with a p.Arg418Gly substitution. ETV6 is a transcriptional repressor that functions through self-oligomerization and is essential for hematopoietic stem cell proliferation and platelet production in mice. Our report was among the first to describe pathogenic germline mutations in ETV6. Functional characterization of the corresponding ETV6 protein mutations showed aberrant cytoplasmic localization, impaired transcriptional repression, and delayed megakaryocyte maturation. Furthermore, we found that mutant ETV6 protein oligomerizes with WT ETV6 and sequesters it from the nucleus suggesting a dominant negative mechanism.
    [Show full text]
  • (NGS) for Primary Endocrine Resistance in Breast Cancer Patients
    Int J Clin Exp Pathol 2018;11(11):5450-5458 www.ijcep.com /ISSN:1936-2625/IJCEP0084102 Original Article Impact of next-generation sequencing (NGS) for primary endocrine resistance in breast cancer patients Ruoyang Li1*, Tiantian Tang1*, Tianli Hui1, Zhenchuan Song1, Fugen Li2, Jingyu Li2, Jiajia Xu2 1Breast Center, Fourth Hospital of Hebei Medical University, Shijiazhuang, China; 2Institute of Precision Medicine, 3D Medicines Inc., Shanghai, China. *Equal contributors. Received August 15, 2018; Accepted September 22, 2018; Epub November 1, 2018; Published November 15, 2018 Abstract: Multiple mechanisms have been detected to account for the acquired resistance to endocrine therapies in breast cancer. In this study we retrospectively studied the mechanism of primary endocrine resistance in estrogen receptor positive (ER+) breast cancer patients by next-generation sequencing (NGS). Tumor specimens and matched blood samples were obtained from 24 ER+ breast cancer patients. Fifteen of them displayed endocrine resistance, including recurrence and/or metastases within 24 months from the beginning of endocrine therapy, and 9 pa- tients remained sensitive to endocrine therapy for more than 5 years. Genomic DNA of tumor tissue was extracted from formalin-fixed paraffin-embedded (FFPE) tumor tissue blocks. Genomic DNA of normal tissue was extracted from peripheral blood mononuclear cells (PBMC). Sequencing libraries for each sample were prepared, followed by target capturing for 372 genes that are frequently rearranged in cancers. Massive parallel
    [Show full text]
  • Pallister-Killian Syndrome Groups Worldwide Who Are Studying the Different Aspects of This Syndrome
    Inform Network Support Rare Chromosome Disorder Support Group G1, The Stables, Station Road West, Oxted, Surrey RH8 9EE, United Kingdom Tel: +44(0)1883 723356 [email protected] I www.rarechromo.org Join Unique for family links, information and support. Unique is a charity without government funding, existing entirely on donations and grants. If you can, please make a donation via our website at Pallister-Killian www.rarechromo.org Please help us to help you! Websites, Facebook groups http://pkskids.net syndrome http://pkskids.ning.com www.pks.org.au www.pksitalia.org www.facebook.com/pages/Pallister-Killian-Syndrome-PKS www.facebook.com/pages/Sindrome-Pallister-Killian (in Spanish) Unique mentions other organisations’ message boards and websites to help families looking for information. This does not imply that we endorse their content or have any responsibility for it. This information guide is not a substitute for personal medical advice. Families should consult a medically qualified clinician in all matters relating to genetic diagnosis, management and health. Information on genetic changes is a very fast-moving field and while the information in this guide is believed to be the best available at the time of publication, some facts may later change. Unique does its best to keep abreast of changing information and to review its published guides as needed. This booklet was compiled by Unique and reviewed by Dr Michel Vekemans, Department of Genetics, Hopital Necker Enfants Malades, Paris, France 2004 and by Unique’s chief medical advisor, Professor Maj Hultén, Professor of Reproductive Genetics, University of Warwick, UK. 2005 (PM).
    [Show full text]