5-Azacytidine prevents transgene methylation in vivo

Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine k...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Gene therapy 1999-04, Vol.6 (4), p.703-707
Hauptverfasser: DI IANNI, M, TERENZI, A, PERRUCCIO, K, CIURNELLI, R, LUCHERONI, F, BENEDETTI, R, MARTELLI, M. F, TABILIO, A
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 707
container_issue 4
container_start_page 703
container_title Gene therapy
container_volume 6
creator DI IANNI, M
TERENZI, A
PERRUCCIO, K
CIURNELLI, R
LUCHERONI, F
BENEDETTI, R
MARTELLI, M. F
TABILIO, A
description Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine kinase suicide gene (tk), which induces sensitivity to ganciclovir (gcv) and through an IRES sequence, the bacterial beta-galactosidase gene (lacZ) as a marker gene. About 90% of the U937 cells expressed the transgene. By injecting the transduced U937 cells in severe combined immunodeficient disease (SCID) mice, we generated a tumor which, during in vivo treatment with aza-C, maintained the high expression of lacZ and tk genes at the baseline values. LacZ-positive cells in the tumour masses after death was weak (1-2%) in the control group, while in mice treated with aza-C it was maintained at 90%. The delay in tumour onset was significantly longer when animals were treated with both aza-C and gcv (P < 0.0001) compared with animals treated with gcv or with aza-C alone. The prevention of silencing phenomena has important implications for gene therapy, because an efficient transduction associated with appropriate drug therapy, might be a powerful strategy for successful application of gene therapy protocols.
doi_str_mv 10.1038/sj.gt.3300848
format Article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_70016218</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2644615506</sourcerecordid><originalsourceid>FETCH-LOGICAL-c416t-b50b2bd6e371af08ac07ddaf29d433e968108c578243ef8ed62b7439798ebc333</originalsourceid><addsrcrecordid>eNqF0EtLAzEUBeAgiq3VpVspKO6m5jVJZinFFxTc6DpkMnfqDPOoSaZQf70pHVDcuMolfBzuPQhdErwgmKk7Xy_WYcEYxoqrIzQlXIok5YIeoynORJZIQtUEnXlfY4y5VPQUTUgcBGV0inia3H8ZuwtVUXUw3zjYQhf8PDjT-TXErxbCx64xoeq7edXNt9W2P0cnpWk8XIzvDL0_Prwtn5PV69PL8n6VWE5ESPIU5zQvBDBJTImVsVgWhSlpVnDGIBOKYGXTuBJnUCooBM0lZ5nMFOSWMTZDt4fcjes_B_BBt5W30DSmg37wWmJMBCXqX0gkS2XG94nXf2DdD66LR2gqOBckTbGIKjko63rvHZR646rWuJ0mWO9b177W66DH1qO_GlOHvIXilz7UHMHNCIy3piljubbyP04KwZhi323LiNo</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2644615506</pqid></control><display><type>article</type><title>5-Azacytidine prevents transgene methylation in vivo</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>DI IANNI, M ; TERENZI, A ; PERRUCCIO, K ; CIURNELLI, R ; LUCHERONI, F ; BENEDETTI, R ; MARTELLI, M. F ; TABILIO, A</creator><creatorcontrib>DI IANNI, M ; TERENZI, A ; PERRUCCIO, K ; CIURNELLI, R ; LUCHERONI, F ; BENEDETTI, R ; MARTELLI, M. F ; TABILIO, A</creatorcontrib><description>Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine kinase suicide gene (tk), which induces sensitivity to ganciclovir (gcv) and through an IRES sequence, the bacterial beta-galactosidase gene (lacZ) as a marker gene. About 90% of the U937 cells expressed the transgene. By injecting the transduced U937 cells in severe combined immunodeficient disease (SCID) mice, we generated a tumor which, during in vivo treatment with aza-C, maintained the high expression of lacZ and tk genes at the baseline values. LacZ-positive cells in the tumour masses after death was weak (1-2%) in the control group, while in mice treated with aza-C it was maintained at 90%. The delay in tumour onset was significantly longer when animals were treated with both aza-C and gcv (P &lt; 0.0001) compared with animals treated with gcv or with aza-C alone. The prevention of silencing phenomena has important implications for gene therapy, because an efficient transduction associated with appropriate drug therapy, might be a powerful strategy for successful application of gene therapy protocols.</description><identifier>ISSN: 0969-7128</identifier><identifier>EISSN: 1476-5462</identifier><identifier>DOI: 10.1038/sj.gt.3300848</identifier><identifier>PMID: 10476232</identifier><language>eng</language><publisher>Basingstoke: Nature Publishing Group</publisher><subject>5-azacytidine ; Animals ; Antiviral Agents - administration &amp; dosage ; Azacitidine - therapeutic use ; Azacytidine ; Biological and medical sciences ; Biotechnology ; DNA Methylation - drug effects ; Drug therapy ; Fundamental and applied biological sciences. Psychology ; Ganciclovir ; Ganciclovir - administration &amp; dosage ; Gene Expression - drug effects ; Gene therapy ; Genetic Therapy - methods ; Genetic Vectors - administration &amp; dosage ; Genetic Vectors - genetics ; Health. Pharmaceutical industry ; Humans ; Immunodeficiency ; Industrial applications and implications. Economical aspects ; Lac Operon - genetics ; Mice ; Mice, SCID ; Retroviridae - genetics ; Severe combined immunodeficiency ; Suicide ; Suicide genes ; Thymidine ; Thymidine kinase ; Thymidine Kinase - genetics ; Transgenes ; Tumors ; U937 Cells ; β-Galactosidase</subject><ispartof>Gene therapy, 1999-04, Vol.6 (4), p.703-707</ispartof><rights>1999 INIST-CNRS</rights><rights>Macmillan Publishers Limited 1999.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c416t-b50b2bd6e371af08ac07ddaf29d433e968108c578243ef8ed62b7439798ebc333</citedby><cites>FETCH-LOGICAL-c416t-b50b2bd6e371af08ac07ddaf29d433e968108c578243ef8ed62b7439798ebc333</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=1766338$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10476232$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>DI IANNI, M</creatorcontrib><creatorcontrib>TERENZI, A</creatorcontrib><creatorcontrib>PERRUCCIO, K</creatorcontrib><creatorcontrib>CIURNELLI, R</creatorcontrib><creatorcontrib>LUCHERONI, F</creatorcontrib><creatorcontrib>BENEDETTI, R</creatorcontrib><creatorcontrib>MARTELLI, M. F</creatorcontrib><creatorcontrib>TABILIO, A</creatorcontrib><title>5-Azacytidine prevents transgene methylation in vivo</title><title>Gene therapy</title><addtitle>Gene Ther</addtitle><description>Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine kinase suicide gene (tk), which induces sensitivity to ganciclovir (gcv) and through an IRES sequence, the bacterial beta-galactosidase gene (lacZ) as a marker gene. About 90% of the U937 cells expressed the transgene. By injecting the transduced U937 cells in severe combined immunodeficient disease (SCID) mice, we generated a tumor which, during in vivo treatment with aza-C, maintained the high expression of lacZ and tk genes at the baseline values. LacZ-positive cells in the tumour masses after death was weak (1-2%) in the control group, while in mice treated with aza-C it was maintained at 90%. The delay in tumour onset was significantly longer when animals were treated with both aza-C and gcv (P &lt; 0.0001) compared with animals treated with gcv or with aza-C alone. The prevention of silencing phenomena has important implications for gene therapy, because an efficient transduction associated with appropriate drug therapy, might be a powerful strategy for successful application of gene therapy protocols.</description><subject>5-azacytidine</subject><subject>Animals</subject><subject>Antiviral Agents - administration &amp; dosage</subject><subject>Azacitidine - therapeutic use</subject><subject>Azacytidine</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>DNA Methylation - drug effects</subject><subject>Drug therapy</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Ganciclovir</subject><subject>Ganciclovir - administration &amp; dosage</subject><subject>Gene Expression - drug effects</subject><subject>Gene therapy</subject><subject>Genetic Therapy - methods</subject><subject>Genetic Vectors - administration &amp; dosage</subject><subject>Genetic Vectors - genetics</subject><subject>Health. Pharmaceutical industry</subject><subject>Humans</subject><subject>Immunodeficiency</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Lac Operon - genetics</subject><subject>Mice</subject><subject>Mice, SCID</subject><subject>Retroviridae - genetics</subject><subject>Severe combined immunodeficiency</subject><subject>Suicide</subject><subject>Suicide genes</subject><subject>Thymidine</subject><subject>Thymidine kinase</subject><subject>Thymidine Kinase - genetics</subject><subject>Transgenes</subject><subject>Tumors</subject><subject>U937 Cells</subject><subject>β-Galactosidase</subject><issn>0969-7128</issn><issn>1476-5462</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0EtLAzEUBeAgiq3VpVspKO6m5jVJZinFFxTc6DpkMnfqDPOoSaZQf70pHVDcuMolfBzuPQhdErwgmKk7Xy_WYcEYxoqrIzQlXIok5YIeoynORJZIQtUEnXlfY4y5VPQUTUgcBGV0inia3H8ZuwtVUXUw3zjYQhf8PDjT-TXErxbCx64xoeq7edXNt9W2P0cnpWk8XIzvDL0_Prwtn5PV69PL8n6VWE5ESPIU5zQvBDBJTImVsVgWhSlpVnDGIBOKYGXTuBJnUCooBM0lZ5nMFOSWMTZDt4fcjes_B_BBt5W30DSmg37wWmJMBCXqX0gkS2XG94nXf2DdD66LR2gqOBckTbGIKjko63rvHZR646rWuJ0mWO9b177W66DH1qO_GlOHvIXilz7UHMHNCIy3piljubbyP04KwZhi323LiNo</recordid><startdate>19990401</startdate><enddate>19990401</enddate><creator>DI IANNI, M</creator><creator>TERENZI, A</creator><creator>PERRUCCIO, K</creator><creator>CIURNELLI, R</creator><creator>LUCHERONI, F</creator><creator>BENEDETTI, R</creator><creator>MARTELLI, M. F</creator><creator>TABILIO, A</creator><general>Nature Publishing Group</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>P64</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>19990401</creationdate><title>5-Azacytidine prevents transgene methylation in vivo</title><author>DI IANNI, M ; TERENZI, A ; PERRUCCIO, K ; CIURNELLI, R ; LUCHERONI, F ; BENEDETTI, R ; MARTELLI, M. F ; TABILIO, A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c416t-b50b2bd6e371af08ac07ddaf29d433e968108c578243ef8ed62b7439798ebc333</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>5-azacytidine</topic><topic>Animals</topic><topic>Antiviral Agents - administration &amp; dosage</topic><topic>Azacitidine - therapeutic use</topic><topic>Azacytidine</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>DNA Methylation - drug effects</topic><topic>Drug therapy</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Ganciclovir</topic><topic>Ganciclovir - administration &amp; dosage</topic><topic>Gene Expression - drug effects</topic><topic>Gene therapy</topic><topic>Genetic Therapy - methods</topic><topic>Genetic Vectors - administration &amp; dosage</topic><topic>Genetic Vectors - genetics</topic><topic>Health. Pharmaceutical industry</topic><topic>Humans</topic><topic>Immunodeficiency</topic><topic>Industrial applications and implications. Economical aspects</topic><topic>Lac Operon - genetics</topic><topic>Mice</topic><topic>Mice, SCID</topic><topic>Retroviridae - genetics</topic><topic>Severe combined immunodeficiency</topic><topic>Suicide</topic><topic>Suicide genes</topic><topic>Thymidine</topic><topic>Thymidine kinase</topic><topic>Thymidine Kinase - genetics</topic><topic>Transgenes</topic><topic>Tumors</topic><topic>U937 Cells</topic><topic>β-Galactosidase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>DI IANNI, M</creatorcontrib><creatorcontrib>TERENZI, A</creatorcontrib><creatorcontrib>PERRUCCIO, K</creatorcontrib><creatorcontrib>CIURNELLI, R</creatorcontrib><creatorcontrib>LUCHERONI, F</creatorcontrib><creatorcontrib>BENEDETTI, R</creatorcontrib><creatorcontrib>MARTELLI, M. F</creatorcontrib><creatorcontrib>TABILIO, A</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium &amp; Calcified Tissue Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Gene therapy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>DI IANNI, M</au><au>TERENZI, A</au><au>PERRUCCIO, K</au><au>CIURNELLI, R</au><au>LUCHERONI, F</au><au>BENEDETTI, R</au><au>MARTELLI, M. F</au><au>TABILIO, A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>5-Azacytidine prevents transgene methylation in vivo</atitle><jtitle>Gene therapy</jtitle><addtitle>Gene Ther</addtitle><date>1999-04-01</date><risdate>1999</risdate><volume>6</volume><issue>4</issue><spage>703</spage><epage>707</epage><pages>703-707</pages><issn>0969-7128</issn><eissn>1476-5462</eissn><abstract>Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine kinase suicide gene (tk), which induces sensitivity to ganciclovir (gcv) and through an IRES sequence, the bacterial beta-galactosidase gene (lacZ) as a marker gene. About 90% of the U937 cells expressed the transgene. By injecting the transduced U937 cells in severe combined immunodeficient disease (SCID) mice, we generated a tumor which, during in vivo treatment with aza-C, maintained the high expression of lacZ and tk genes at the baseline values. LacZ-positive cells in the tumour masses after death was weak (1-2%) in the control group, while in mice treated with aza-C it was maintained at 90%. The delay in tumour onset was significantly longer when animals were treated with both aza-C and gcv (P &lt; 0.0001) compared with animals treated with gcv or with aza-C alone. The prevention of silencing phenomena has important implications for gene therapy, because an efficient transduction associated with appropriate drug therapy, might be a powerful strategy for successful application of gene therapy protocols.</abstract><cop>Basingstoke</cop><pub>Nature Publishing Group</pub><pmid>10476232</pmid><doi>10.1038/sj.gt.3300848</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0969-7128
ispartof Gene therapy, 1999-04, Vol.6 (4), p.703-707
issn 0969-7128
1476-5462
language eng
recordid cdi_proquest_miscellaneous_70016218
source MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection
subjects 5-azacytidine
Animals
Antiviral Agents - administration & dosage
Azacitidine - therapeutic use
Azacytidine
Biological and medical sciences
Biotechnology
DNA Methylation - drug effects
Drug therapy
Fundamental and applied biological sciences. Psychology
Ganciclovir
Ganciclovir - administration & dosage
Gene Expression - drug effects
Gene therapy
Genetic Therapy - methods
Genetic Vectors - administration & dosage
Genetic Vectors - genetics
Health. Pharmaceutical industry
Humans
Immunodeficiency
Industrial applications and implications. Economical aspects
Lac Operon - genetics
Mice
Mice, SCID
Retroviridae - genetics
Severe combined immunodeficiency
Suicide
Suicide genes
Thymidine
Thymidine kinase
Thymidine Kinase - genetics
Transgenes
Tumors
U937 Cells
β-Galactosidase
title 5-Azacytidine prevents transgene methylation in vivo
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T00%3A40%3A34IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=5-Azacytidine%20prevents%20transgene%20methylation%20in%20vivo&rft.jtitle=Gene%20therapy&rft.au=DI%20IANNI,%20M&rft.date=1999-04-01&rft.volume=6&rft.issue=4&rft.spage=703&rft.epage=707&rft.pages=703-707&rft.issn=0969-7128&rft.eissn=1476-5462&rft_id=info:doi/10.1038/sj.gt.3300848&rft_dat=%3Cproquest_cross%3E2644615506%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2644615506&rft_id=info:pmid/10476232&rfr_iscdi=true