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Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro
Osteosarcoma is the most frequent malignant disease of the bone. On the basis of early clinical experience in the 1960s with H-1 protoparvovirus (H-1PV) in osteosarcoma patients, this effective oncolytic virus was selected for systematic preclinical testing on various osteosarcoma cell cultures. A p...
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Published in: | Viruses 2017-10, Vol.9 (10), p.301 |
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description | Osteosarcoma is the most frequent malignant disease of the bone. On the basis of early clinical experience in the 1960s with H-1 protoparvovirus (H-1PV) in osteosarcoma patients, this effective oncolytic virus was selected for systematic preclinical testing on various osteosarcoma cell cultures. A panel of five human osteosarcoma cell lines (CAL 72, H-OS, MG-63, SaOS-2, U-2OS) was tested. Virus oncoselectivity was confirmed by infecting non-malignant human neonatal fibroblasts and osteoblasts used as culture models of non-transformed mesenchymal cells. H-1PV was found to enter osteosarcoma cells and to induce viral DNA replication, transcription of viral genes, and translation to viral proteins. After H-1PV infection, release of infectious viral particles from osteosarcoma cells into the supernatant indicated successful viral assembly and egress. Crystal violet staining revealed progressive cytomorphological changes in all osteosarcoma cell lines. Infection of osteosarcoma cell lines with the standard H-1PV caused an arrest of the cell cycle in the G2 phase, and these lines had a limited capacity for standard H-1PV virus replication. The cytotoxicity of wild-type H-1PV virus towards osteosarcoma cells was compared in vitro with that of two variants, Del H-1PV and DM H-1PV, previously described as fitness variants displaying higher infectivity and spreading in human transformed cell lines of different origins. Surprisingly, wild-type H-1PV displayed the strongest cytostatic and cytotoxic effects in this analysis and thus seems the most promising for the next preclinical validation steps in vivo. |
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On the basis of early clinical experience in the 1960s with H-1 protoparvovirus (H-1PV) in osteosarcoma patients, this effective oncolytic virus was selected for systematic preclinical testing on various osteosarcoma cell cultures. A panel of five human osteosarcoma cell lines (CAL 72, H-OS, MG-63, SaOS-2, U-2OS) was tested. Virus oncoselectivity was confirmed by infecting non-malignant human neonatal fibroblasts and osteoblasts used as culture models of non-transformed mesenchymal cells. H-1PV was found to enter osteosarcoma cells and to induce viral DNA replication, transcription of viral genes, and translation to viral proteins. After H-1PV infection, release of infectious viral particles from osteosarcoma cells into the supernatant indicated successful viral assembly and egress. Crystal violet staining revealed progressive cytomorphological changes in all osteosarcoma cell lines. Infection of osteosarcoma cell lines with the standard H-1PV caused an arrest of the cell cycle in the G2 phase, and these lines had a limited capacity for standard H-1PV virus replication. The cytotoxicity of wild-type H-1PV virus towards osteosarcoma cells was compared in vitro with that of two variants, Del H-1PV and DM H-1PV, previously described as fitness variants displaying higher infectivity and spreading in human transformed cell lines of different origins. Surprisingly, wild-type H-1PV displayed the strongest cytostatic and cytotoxic effects in this analysis and thus seems the most promising for the next preclinical validation steps in vivo.</description><identifier>ISSN: 1999-4915</identifier><identifier>EISSN: 1999-4915</identifier><identifier>DOI: 10.3390/v9100301</identifier><identifier>PMID: 29039746</identifier><language>eng</language><publisher>Switzerland: MDPI AG</publisher><subject>Bone cancer ; Cell culture ; Cell Cycle ; Cell Death ; Cell Line ; Cell Line, Tumor ; Cell lines ; Cytotoxicity ; DNA biosynthesis ; Early experience ; Fibroblasts ; Fitness ; G2 phase ; H-1 parvovirus - physiology ; Humans ; Infectivity ; Lysis ; Mesenchyme ; Neonates ; Oncolysis ; Oncolytic Virotherapy ; oncolytic virus ; Oncolytic Viruses - physiology ; Osteoblasts ; Osteosarcoma ; Osteosarcoma - pathology ; Osteosarcoma - virology ; Osteosarcoma cells ; Parvoviruses ; protoparvovirus H-1 (H-1PV) ; Replication ; Sarcoma ; Transcription ; viral cytotoxicity ; Virus Replication</subject><ispartof>Viruses, 2017-10, Vol.9 (10), p.301</ispartof><rights>Copyright MDPI AG 2017</rights><rights>2017 by the authors. 2017</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c466t-65270bbb5060887d0399ca27360d12948447da4df672fff7e43f66c3a09496fc3</citedby><cites>FETCH-LOGICAL-c466t-65270bbb5060887d0399ca27360d12948447da4df672fff7e43f66c3a09496fc3</cites><orcidid>0000-0001-9572-2476 ; 0000-0002-6598-8369 ; 0000-0002-2233-3346</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/1965583815/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/1965583815?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793,75126</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29039746$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Geiss, Carsten</creatorcontrib><creatorcontrib>Kis, Zoltán</creatorcontrib><creatorcontrib>Leuchs, Barbara</creatorcontrib><creatorcontrib>Frank-Stöhr, Monika</creatorcontrib><creatorcontrib>Schlehofer, Jörg R</creatorcontrib><creatorcontrib>Rommelaere, Jean</creatorcontrib><creatorcontrib>Dinsart, Christiane</creatorcontrib><creatorcontrib>Lacroix, Jeannine</creatorcontrib><title>Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro</title><title>Viruses</title><addtitle>Viruses</addtitle><description>Osteosarcoma is the most frequent malignant disease of the bone. On the basis of early clinical experience in the 1960s with H-1 protoparvovirus (H-1PV) in osteosarcoma patients, this effective oncolytic virus was selected for systematic preclinical testing on various osteosarcoma cell cultures. A panel of five human osteosarcoma cell lines (CAL 72, H-OS, MG-63, SaOS-2, U-2OS) was tested. Virus oncoselectivity was confirmed by infecting non-malignant human neonatal fibroblasts and osteoblasts used as culture models of non-transformed mesenchymal cells. H-1PV was found to enter osteosarcoma cells and to induce viral DNA replication, transcription of viral genes, and translation to viral proteins. After H-1PV infection, release of infectious viral particles from osteosarcoma cells into the supernatant indicated successful viral assembly and egress. Crystal violet staining revealed progressive cytomorphological changes in all osteosarcoma cell lines. Infection of osteosarcoma cell lines with the standard H-1PV caused an arrest of the cell cycle in the G2 phase, and these lines had a limited capacity for standard H-1PV virus replication. The cytotoxicity of wild-type H-1PV virus towards osteosarcoma cells was compared in vitro with that of two variants, Del H-1PV and DM H-1PV, previously described as fitness variants displaying higher infectivity and spreading in human transformed cell lines of different origins. 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On the basis of early clinical experience in the 1960s with H-1 protoparvovirus (H-1PV) in osteosarcoma patients, this effective oncolytic virus was selected for systematic preclinical testing on various osteosarcoma cell cultures. A panel of five human osteosarcoma cell lines (CAL 72, H-OS, MG-63, SaOS-2, U-2OS) was tested. Virus oncoselectivity was confirmed by infecting non-malignant human neonatal fibroblasts and osteoblasts used as culture models of non-transformed mesenchymal cells. H-1PV was found to enter osteosarcoma cells and to induce viral DNA replication, transcription of viral genes, and translation to viral proteins. After H-1PV infection, release of infectious viral particles from osteosarcoma cells into the supernatant indicated successful viral assembly and egress. Crystal violet staining revealed progressive cytomorphological changes in all osteosarcoma cell lines. Infection of osteosarcoma cell lines with the standard H-1PV caused an arrest of the cell cycle in the G2 phase, and these lines had a limited capacity for standard H-1PV virus replication. The cytotoxicity of wild-type H-1PV virus towards osteosarcoma cells was compared in vitro with that of two variants, Del H-1PV and DM H-1PV, previously described as fitness variants displaying higher infectivity and spreading in human transformed cell lines of different origins. Surprisingly, wild-type H-1PV displayed the strongest cytostatic and cytotoxic effects in this analysis and thus seems the most promising for the next preclinical validation steps in vivo.</abstract><cop>Switzerland</cop><pub>MDPI AG</pub><pmid>29039746</pmid><doi>10.3390/v9100301</doi><orcidid>https://orcid.org/0000-0001-9572-2476</orcidid><orcidid>https://orcid.org/0000-0002-6598-8369</orcidid><orcidid>https://orcid.org/0000-0002-2233-3346</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Bone cancer Cell culture Cell Cycle Cell Death Cell Line Cell Line, Tumor Cell lines Cytotoxicity DNA biosynthesis Early experience Fibroblasts Fitness G2 phase H-1 parvovirus - physiology Humans Infectivity Lysis Mesenchyme Neonates Oncolysis Oncolytic Virotherapy oncolytic virus Oncolytic Viruses - physiology Osteoblasts Osteosarcoma Osteosarcoma - pathology Osteosarcoma - virology Osteosarcoma cells Parvoviruses protoparvovirus H-1 (H-1PV) Replication Sarcoma Transcription viral cytotoxicity Virus Replication |
title | Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro |
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