Transfection efficiency and toxicity following delivery of naked plasmid DNA and cationic lipid–DNA complexes to ovine lung segments
We defined, using a novel large animal model system, the acute pathologic response to localized pulmonary administration of either naked plasmid DNA (pDNA) or cationic lipid–pDNA complexes (pDNA:GL67) and related such responses to concomitant indicators of transfection efficiency, namely levels of c...
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| Veröffentlicht in: | Molecular therapy 2003-10, Vol.8 (4), p.646-653 |
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| creator | Emerson, Michael Renwick, Louise Tate, Stephen Rhind, Susan Milne, Elspeth Painter (née Alsop), Hazel Boyd, A.Christopher McLachlan, Gerry Griesenbach, Uta Cheng, Seng H Gill, Deborah R Hyde, Stephen C Baker, Alison Alton, Eric W Porteous, David J Collie, D.David S |
| description | We defined, using a novel large animal model system, the acute pathologic response to localized pulmonary administration of either naked plasmid DNA (pDNA) or cationic lipid–pDNA complexes (pDNA:GL67) and related such responses to concomitant indicators of transfection efficiency, namely levels of chloramphenicol acetyl transferase (CAT) protein and mRNA in specific lung tissue compartments. We instilled doses of 0.2, 1, and 5 mg pDNA to spatially distinct lung segments in six anesthetized sheep and doses of 0.2, 1, and 5 mg pDNA:GL67 to a further six sheep. Twenty-four hours after gene delivery the sheep were euthanized and necropsy examination with sampling of relevant tissues was carried out. Levels of plasmid-derived CAT-specific mRNA and CAT protein in samples derived from segments treated with either pDNA or pDNA:GL67 increased in relation to the administered dose. Levels of mRNA and protein expression were greater for pDNA:GL67 than for pDNA alone. A significant correlation was observed between mRNA and protein expression in samples derived from airways treated with pDNA:GL67. Histopathological changes following administration of both pDNA and pDNA:GL67 were characterized by a neutrophilic inflammation predominantly oriented on airways. The severity of the inflammatory response appeared to correlate with the administered dose of DNA and was generally more severe for pDNA:GL67. |
| doi_str_mv | 10.1016/S1525-0016(03)00233-8 |
| format | Article |
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We instilled doses of 0.2, 1, and 5 mg pDNA to spatially distinct lung segments in six anesthetized sheep and doses of 0.2, 1, and 5 mg pDNA:GL67 to a further six sheep. Twenty-four hours after gene delivery the sheep were euthanized and necropsy examination with sampling of relevant tissues was carried out. Levels of plasmid-derived CAT-specific mRNA and CAT protein in samples derived from segments treated with either pDNA or pDNA:GL67 increased in relation to the administered dose. Levels of mRNA and protein expression were greater for pDNA:GL67 than for pDNA alone. A significant correlation was observed between mRNA and protein expression in samples derived from airways treated with pDNA:GL67. Histopathological changes following administration of both pDNA and pDNA:GL67 were characterized by a neutrophilic inflammation predominantly oriented on airways. The severity of the inflammatory response appeared to correlate with the administered dose of DNA and was generally more severe for pDNA:GL67.</description><identifier>ISSN: 1525-0016</identifier><identifier>EISSN: 1525-0024</identifier><identifier>DOI: 10.1016/S1525-0016(03)00233-8</identifier><identifier>PMID: 14529838</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; cationic lipid ; chloramphenicol acetyltransferase ; Chloramphenicol O-Acetyltransferase - genetics ; Chloramphenicol O-Acetyltransferase - metabolism ; Clinical trials ; Cystic fibrosis ; domestic ; Drug dosages ; Gene therapy ; gene transfer ; genetic vectors ; Lipids ; Lipids - pharmacology ; Lipids - toxicity ; lung ; Lung - drug effects ; Lung - pathology ; messenger ; Plasmids - pharmacology ; Plasmids - toxicity ; Protein expression ; Proteins ; RNA ; Sheep ; Sheep - genetics ; Sheep - metabolism ; Toxicity ; toxicology ; Transfection ; Veterinary medicine</subject><ispartof>Molecular therapy, 2003-10, Vol.8 (4), p.646-653</ispartof><rights>2003 The American Society of Gene Therapy</rights><rights>Copyright Nature Publishing Group Oct 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c462t-79e47936118f4e6036e44eb945e4fc0621f78fdb420e27efc7f70a688d3bda3</citedby><cites>FETCH-LOGICAL-c462t-79e47936118f4e6036e44eb945e4fc0621f78fdb420e27efc7f70a688d3bda3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.proquest.com/docview/1792822156?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,64385,64389,72469</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14529838$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Emerson, Michael</creatorcontrib><creatorcontrib>Renwick, Louise</creatorcontrib><creatorcontrib>Tate, Stephen</creatorcontrib><creatorcontrib>Rhind, Susan</creatorcontrib><creatorcontrib>Milne, Elspeth</creatorcontrib><creatorcontrib>Painter (née Alsop), Hazel</creatorcontrib><creatorcontrib>Boyd, A.Christopher</creatorcontrib><creatorcontrib>McLachlan, Gerry</creatorcontrib><creatorcontrib>Griesenbach, Uta</creatorcontrib><creatorcontrib>Cheng, Seng H</creatorcontrib><creatorcontrib>Gill, Deborah R</creatorcontrib><creatorcontrib>Hyde, Stephen C</creatorcontrib><creatorcontrib>Baker, Alison</creatorcontrib><creatorcontrib>Alton, Eric W</creatorcontrib><creatorcontrib>Porteous, David J</creatorcontrib><creatorcontrib>Collie, D.David S</creatorcontrib><title>Transfection efficiency and toxicity following delivery of naked plasmid DNA and cationic lipid–DNA complexes to ovine lung segments</title><title>Molecular therapy</title><addtitle>Mol Ther</addtitle><description>We defined, using a novel large animal model system, the acute pathologic response to localized pulmonary administration of either naked plasmid DNA (pDNA) or cationic lipid–pDNA complexes (pDNA:GL67) and related such responses to concomitant indicators of transfection efficiency, namely levels of chloramphenicol acetyl transferase (CAT) protein and mRNA in specific lung tissue compartments. We instilled doses of 0.2, 1, and 5 mg pDNA to spatially distinct lung segments in six anesthetized sheep and doses of 0.2, 1, and 5 mg pDNA:GL67 to a further six sheep. Twenty-four hours after gene delivery the sheep were euthanized and necropsy examination with sampling of relevant tissues was carried out. Levels of plasmid-derived CAT-specific mRNA and CAT protein in samples derived from segments treated with either pDNA or pDNA:GL67 increased in relation to the administered dose. Levels of mRNA and protein expression were greater for pDNA:GL67 than for pDNA alone. A significant correlation was observed between mRNA and protein expression in samples derived from airways treated with pDNA:GL67. Histopathological changes following administration of both pDNA and pDNA:GL67 were characterized by a neutrophilic inflammation predominantly oriented on airways. The severity of the inflammatory response appeared to correlate with the administered dose of DNA and was generally more severe for pDNA:GL67.</description><subject>Animals</subject><subject>cationic lipid</subject><subject>chloramphenicol acetyltransferase</subject><subject>Chloramphenicol O-Acetyltransferase - genetics</subject><subject>Chloramphenicol O-Acetyltransferase - metabolism</subject><subject>Clinical trials</subject><subject>Cystic fibrosis</subject><subject>domestic</subject><subject>Drug dosages</subject><subject>Gene therapy</subject><subject>gene transfer</subject><subject>genetic vectors</subject><subject>Lipids</subject><subject>Lipids - pharmacology</subject><subject>Lipids - toxicity</subject><subject>lung</subject><subject>Lung - drug effects</subject><subject>Lung - pathology</subject><subject>messenger</subject><subject>Plasmids - pharmacology</subject><subject>Plasmids - toxicity</subject><subject>Protein expression</subject><subject>Proteins</subject><subject>RNA</subject><subject>Sheep</subject><subject>Sheep - genetics</subject><subject>Sheep - metabolism</subject><subject>Toxicity</subject><subject>toxicology</subject><subject>Transfection</subject><subject>Veterinary 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Michael ; Renwick, Louise ; Tate, Stephen ; Rhind, Susan ; Milne, Elspeth ; Painter (née Alsop), Hazel ; Boyd, A.Christopher ; McLachlan, Gerry ; Griesenbach, Uta ; Cheng, Seng H ; Gill, Deborah R ; Hyde, Stephen C ; Baker, Alison ; Alton, Eric W ; Porteous, David J ; Collie, D.David S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c462t-79e47936118f4e6036e44eb945e4fc0621f78fdb420e27efc7f70a688d3bda3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>cationic lipid</topic><topic>chloramphenicol acetyltransferase</topic><topic>Chloramphenicol O-Acetyltransferase - genetics</topic><topic>Chloramphenicol O-Acetyltransferase - metabolism</topic><topic>Clinical trials</topic><topic>Cystic fibrosis</topic><topic>domestic</topic><topic>Drug dosages</topic><topic>Gene therapy</topic><topic>gene transfer</topic><topic>genetic vectors</topic><topic>Lipids</topic><topic>Lipids - pharmacology</topic><topic>Lipids - toxicity</topic><topic>lung</topic><topic>Lung - drug effects</topic><topic>Lung - pathology</topic><topic>messenger</topic><topic>Plasmids - pharmacology</topic><topic>Plasmids - toxicity</topic><topic>Protein expression</topic><topic>Proteins</topic><topic>RNA</topic><topic>Sheep</topic><topic>Sheep - genetics</topic><topic>Sheep - metabolism</topic><topic>Toxicity</topic><topic>toxicology</topic><topic>Transfection</topic><topic>Veterinary medicine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Emerson, Michael</creatorcontrib><creatorcontrib>Renwick, Louise</creatorcontrib><creatorcontrib>Tate, Stephen</creatorcontrib><creatorcontrib>Rhind, Susan</creatorcontrib><creatorcontrib>Milne, Elspeth</creatorcontrib><creatorcontrib>Painter (née Alsop), Hazel</creatorcontrib><creatorcontrib>Boyd, A.Christopher</creatorcontrib><creatorcontrib>McLachlan, Gerry</creatorcontrib><creatorcontrib>Griesenbach, Uta</creatorcontrib><creatorcontrib>Cheng, Seng H</creatorcontrib><creatorcontrib>Gill, Deborah R</creatorcontrib><creatorcontrib>Hyde, Stephen C</creatorcontrib><creatorcontrib>Baker, Alison</creatorcontrib><creatorcontrib>Alton, Eric W</creatorcontrib><creatorcontrib>Porteous, David J</creatorcontrib><creatorcontrib>Collie, D.David S</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni 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Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><jtitle>Molecular therapy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Emerson, Michael</au><au>Renwick, Louise</au><au>Tate, Stephen</au><au>Rhind, Susan</au><au>Milne, Elspeth</au><au>Painter (née Alsop), Hazel</au><au>Boyd, A.Christopher</au><au>McLachlan, Gerry</au><au>Griesenbach, Uta</au><au>Cheng, Seng H</au><au>Gill, Deborah R</au><au>Hyde, Stephen C</au><au>Baker, Alison</au><au>Alton, Eric W</au><au>Porteous, David J</au><au>Collie, D.David S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transfection efficiency and toxicity following delivery of naked plasmid DNA and cationic lipid–DNA complexes to ovine lung segments</atitle><jtitle>Molecular therapy</jtitle><addtitle>Mol Ther</addtitle><date>2003-10-01</date><risdate>2003</risdate><volume>8</volume><issue>4</issue><spage>646</spage><epage>653</epage><pages>646-653</pages><issn>1525-0016</issn><eissn>1525-0024</eissn><abstract>We defined, using a novel large animal model system, the acute pathologic response to localized pulmonary administration of either naked plasmid DNA (pDNA) or cationic lipid–pDNA complexes (pDNA:GL67) and related such responses to concomitant indicators of transfection efficiency, namely levels of chloramphenicol acetyl transferase (CAT) protein and mRNA in specific lung tissue compartments. We instilled doses of 0.2, 1, and 5 mg pDNA to spatially distinct lung segments in six anesthetized sheep and doses of 0.2, 1, and 5 mg pDNA:GL67 to a further six sheep. Twenty-four hours after gene delivery the sheep were euthanized and necropsy examination with sampling of relevant tissues was carried out. Levels of plasmid-derived CAT-specific mRNA and CAT protein in samples derived from segments treated with either pDNA or pDNA:GL67 increased in relation to the administered dose. Levels of mRNA and protein expression were greater for pDNA:GL67 than for pDNA alone. A significant correlation was observed between mRNA and protein expression in samples derived from airways treated with pDNA:GL67. Histopathological changes following administration of both pDNA and pDNA:GL67 were characterized by a neutrophilic inflammation predominantly oriented on airways. The severity of the inflammatory response appeared to correlate with the administered dose of DNA and was generally more severe for pDNA:GL67.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>14529838</pmid><doi>10.1016/S1525-0016(03)00233-8</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals cationic lipid chloramphenicol acetyltransferase Chloramphenicol O-Acetyltransferase - genetics Chloramphenicol O-Acetyltransferase - metabolism Clinical trials Cystic fibrosis domestic Drug dosages Gene therapy gene transfer genetic vectors Lipids Lipids - pharmacology Lipids - toxicity lung Lung - drug effects Lung - pathology messenger Plasmids - pharmacology Plasmids - toxicity Protein expression Proteins RNA Sheep Sheep - genetics Sheep - metabolism Toxicity toxicology Transfection Veterinary medicine |
| title | Transfection efficiency and toxicity following delivery of naked plasmid DNA and cationic lipid–DNA complexes to ovine lung segments |
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