Ocular cell transfection with the human basic fibroblast growth factor gene delays photoreceptor cell degeneration in RCS rats
Based on the K8/JTS-1-mediated transfection technique, we developed an in vivo protocol for an efficient transfer of plasmid DNA to ocular cells. As determined with condensed plasmids containing reporter genes for either beta-galactosidase (pcDNA-lacZ) or enhanced green fluorescent protein (pREP-EGF...
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| Veröffentlicht in: | Human gene therapy 2000-09, Vol.11 (13), p.1875-1890 |
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| creator | NEUNER-JEHLE, Martin VAN DEN BERGHE, Loic ABITBOL, Marc BONNEL, Sébastien UTEZA, Yves BENMEZIANE, Farid ROUILLOT, Jean-Sébastien MARCHANT, Dominique KOBETZ, Alexandra DUFIER, Jean-Louis MENASCHE, Maurice |
| description | Based on the K8/JTS-1-mediated transfection technique, we developed an in vivo protocol for an efficient transfer of plasmid DNA to ocular cells. As determined with condensed plasmids containing reporter genes for either beta-galactosidase (pcDNA-lacZ) or enhanced green fluorescent protein (pREP-EGFP), the immortalized human retinal epithelial cells RPE D407 and human embryonic kidney 293 cells can be transfected with typical efficiencies of 11 and 19%, respectively. Unlike 293 cells, RPE D407 cells had a reduced viability on transfection with both plasmids. In vivo, subretinal injections of DNA-K8/JTS-1 complexes revealed reporter gene expression in choroidal and RPE cells of normal pink-eyed Royal College of Surgeons (RCS) rats. The validity of this transfection technique in terms of retinal cell survival in RCS rats was then examined by using pREP-hFGF2 plasmid, which encodes the human basic fibroblast growth factor isoforms (hFGF2). Subretinal injection of pREP-hFGF2-K8/JTS-1 complexes into 3-week-old dystrophic RCS rat eyes reveals a delayed photoreceptor cell degeneration 60 days postinjection. In this case, the average analyzed field points with delayed cell dystrophy represent 14 to 17% of the retinal surface as compared with 2.6 and 4% in pREP5beta and vehicle-injected eyes, respectively. Peptide-mediated in oculo transfection thus appears to be a promising technique for the treatment of retinal cell and photoreceptor degenerations. |
| doi_str_mv | 10.1089/10430340050129495 |
| format | Article |
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As determined with condensed plasmids containing reporter genes for either beta-galactosidase (pcDNA-lacZ) or enhanced green fluorescent protein (pREP-EGFP), the immortalized human retinal epithelial cells RPE D407 and human embryonic kidney 293 cells can be transfected with typical efficiencies of 11 and 19%, respectively. Unlike 293 cells, RPE D407 cells had a reduced viability on transfection with both plasmids. In vivo, subretinal injections of DNA-K8/JTS-1 complexes revealed reporter gene expression in choroidal and RPE cells of normal pink-eyed Royal College of Surgeons (RCS) rats. The validity of this transfection technique in terms of retinal cell survival in RCS rats was then examined by using pREP-hFGF2 plasmid, which encodes the human basic fibroblast growth factor isoforms (hFGF2). Subretinal injection of pREP-hFGF2-K8/JTS-1 complexes into 3-week-old dystrophic RCS rat eyes reveals a delayed photoreceptor cell degeneration 60 days postinjection. In this case, the average analyzed field points with delayed cell dystrophy represent 14 to 17% of the retinal surface as compared with 2.6 and 4% in pREP5beta and vehicle-injected eyes, respectively. Peptide-mediated in oculo transfection thus appears to be a promising technique for the treatment of retinal cell and photoreceptor degenerations.</description><identifier>ISSN: 1043-0342</identifier><identifier>EISSN: 1557-7422</identifier><identifier>DOI: 10.1089/10430340050129495</identifier><identifier>PMID: 10986560</identifier><identifier>CODEN: HGTHE3</identifier><language>eng</language><publisher>Larchmont, NY: Liebert</publisher><subject>Animals ; Base Sequence ; beta-Galactosidase - genetics ; beta-Galactosidase - metabolism ; Biological and medical sciences ; Biotechnology ; Cell Survival - genetics ; Fibroblast Growth Factor 2 - genetics ; Fibroblast Growth Factor 2 - metabolism ; Fibroblast Growth Factor 2 - pharmacology ; Fundamental and applied biological sciences. Psychology ; Gene therapy ; Genetic Therapy - methods ; Green Fluorescent Proteins ; Health. Pharmaceutical industry ; Humans ; Industrial applications and implications. Economical aspects ; Luminescent Proteins - genetics ; Luminescent Proteins - metabolism ; Microinjections ; Molecular Sequence Data ; Photoreceptor Cells, Vertebrate - cytology ; Photoreceptor Cells, Vertebrate - pathology ; Photoreceptor Cells, Vertebrate - physiology ; Pigment Epithelium of Eye - cytology ; Plasmids - genetics ; Rats ; Retinal Degeneration - genetics ; Transfection - methods</subject><ispartof>Human gene therapy, 2000-09, Vol.11 (13), p.1875-1890</ispartof><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c356t-3e9fe2782743c8cbb0174fe4f5c4d372584a2b6ff6fa31376ea2738f2d67300d3</citedby><cites>FETCH-LOGICAL-c356t-3e9fe2782743c8cbb0174fe4f5c4d372584a2b6ff6fa31376ea2738f2d67300d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3028,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=806281$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10986560$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>NEUNER-JEHLE, Martin</creatorcontrib><creatorcontrib>VAN DEN BERGHE, Loic</creatorcontrib><creatorcontrib>ABITBOL, Marc</creatorcontrib><creatorcontrib>BONNEL, Sébastien</creatorcontrib><creatorcontrib>UTEZA, Yves</creatorcontrib><creatorcontrib>BENMEZIANE, Farid</creatorcontrib><creatorcontrib>ROUILLOT, Jean-Sébastien</creatorcontrib><creatorcontrib>MARCHANT, Dominique</creatorcontrib><creatorcontrib>KOBETZ, Alexandra</creatorcontrib><creatorcontrib>DUFIER, Jean-Louis</creatorcontrib><creatorcontrib>MENASCHE, Maurice</creatorcontrib><title>Ocular cell transfection with the human basic fibroblast growth factor gene delays photoreceptor cell degeneration in RCS rats</title><title>Human gene therapy</title><addtitle>Hum Gene Ther</addtitle><description>Based on the K8/JTS-1-mediated transfection technique, we developed an in vivo protocol for an efficient transfer of plasmid DNA to ocular cells. As determined with condensed plasmids containing reporter genes for either beta-galactosidase (pcDNA-lacZ) or enhanced green fluorescent protein (pREP-EGFP), the immortalized human retinal epithelial cells RPE D407 and human embryonic kidney 293 cells can be transfected with typical efficiencies of 11 and 19%, respectively. Unlike 293 cells, RPE D407 cells had a reduced viability on transfection with both plasmids. In vivo, subretinal injections of DNA-K8/JTS-1 complexes revealed reporter gene expression in choroidal and RPE cells of normal pink-eyed Royal College of Surgeons (RCS) rats. The validity of this transfection technique in terms of retinal cell survival in RCS rats was then examined by using pREP-hFGF2 plasmid, which encodes the human basic fibroblast growth factor isoforms (hFGF2). Subretinal injection of pREP-hFGF2-K8/JTS-1 complexes into 3-week-old dystrophic RCS rat eyes reveals a delayed photoreceptor cell degeneration 60 days postinjection. In this case, the average analyzed field points with delayed cell dystrophy represent 14 to 17% of the retinal surface as compared with 2.6 and 4% in pREP5beta and vehicle-injected eyes, respectively. Peptide-mediated in oculo transfection thus appears to be a promising technique for the treatment of retinal cell and photoreceptor degenerations.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>beta-Galactosidase - genetics</subject><subject>beta-Galactosidase - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cell Survival - genetics</subject><subject>Fibroblast Growth Factor 2 - genetics</subject><subject>Fibroblast Growth Factor 2 - metabolism</subject><subject>Fibroblast Growth Factor 2 - pharmacology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene therapy</subject><subject>Genetic Therapy - methods</subject><subject>Green Fluorescent Proteins</subject><subject>Health. Pharmaceutical industry</subject><subject>Humans</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Luminescent Proteins - genetics</subject><subject>Luminescent Proteins - metabolism</subject><subject>Microinjections</subject><subject>Molecular Sequence Data</subject><subject>Photoreceptor Cells, Vertebrate - cytology</subject><subject>Photoreceptor Cells, Vertebrate - pathology</subject><subject>Photoreceptor Cells, Vertebrate - physiology</subject><subject>Pigment Epithelium of Eye - cytology</subject><subject>Plasmids - genetics</subject><subject>Rats</subject><subject>Retinal Degeneration - genetics</subject><subject>Transfection - methods</subject><issn>1043-0342</issn><issn>1557-7422</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtLw0AUhQdRrFZ_gBsZENxF5z3JUoovKBR8rMNkcqeNpEmdSSjd-NudtEUEF67u67uHAwehC0puKEmzW0oEJ1wQIgllmcjkATqhUupEC8YOYx_vSQTYCJ2G8EEI5VLpYzSiJEuVVOQEfc1sXxuPLdQ17rxpggPbVW2D11W3wN0C8KJfmgYXJlQWu6rwbVGb0OG5b9eRcMZ2rcdzaACXUJtNwKtFG1dgYTVctsolDIA3W-WqwS-TVxyncIaOnKkDnO_rGL0_3L9NnpLp7PF5cjdNbHTcJRwyB0ynTAtuU1sUhGrhQDhpRck1k6kwrFDOKWc45VqBYZqnjpVKc0JKPkbXO92Vbz97CF2-rMLgzDTQ9iHXjMlMMfUvSLVimjIZQboDrW9D8ODyla-Wxm9ySvIhnfxPOvHnci_eF0sof33s4ojA1R4wwZraxTxsFX64lCiWUv4NrQeX1A</recordid><startdate>20000901</startdate><enddate>20000901</enddate><creator>NEUNER-JEHLE, Martin</creator><creator>VAN DEN BERGHE, Loic</creator><creator>ABITBOL, Marc</creator><creator>BONNEL, Sébastien</creator><creator>UTEZA, Yves</creator><creator>BENMEZIANE, Farid</creator><creator>ROUILLOT, Jean-Sébastien</creator><creator>MARCHANT, Dominique</creator><creator>KOBETZ, Alexandra</creator><creator>DUFIER, Jean-Louis</creator><creator>MENASCHE, Maurice</creator><general>Liebert</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20000901</creationdate><title>Ocular cell transfection with the human basic fibroblast growth factor gene delays photoreceptor cell degeneration in RCS rats</title><author>NEUNER-JEHLE, Martin ; VAN DEN BERGHE, Loic ; ABITBOL, Marc ; BONNEL, Sébastien ; UTEZA, Yves ; BENMEZIANE, Farid ; ROUILLOT, Jean-Sébastien ; MARCHANT, Dominique ; KOBETZ, Alexandra ; DUFIER, Jean-Louis ; MENASCHE, Maurice</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c356t-3e9fe2782743c8cbb0174fe4f5c4d372584a2b6ff6fa31376ea2738f2d67300d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>beta-Galactosidase - genetics</topic><topic>beta-Galactosidase - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cell Survival - genetics</topic><topic>Fibroblast Growth Factor 2 - genetics</topic><topic>Fibroblast Growth Factor 2 - metabolism</topic><topic>Fibroblast Growth Factor 2 - pharmacology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene therapy</topic><topic>Genetic Therapy - methods</topic><topic>Green Fluorescent Proteins</topic><topic>Health. Pharmaceutical industry</topic><topic>Humans</topic><topic>Industrial applications and implications. 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As determined with condensed plasmids containing reporter genes for either beta-galactosidase (pcDNA-lacZ) or enhanced green fluorescent protein (pREP-EGFP), the immortalized human retinal epithelial cells RPE D407 and human embryonic kidney 293 cells can be transfected with typical efficiencies of 11 and 19%, respectively. Unlike 293 cells, RPE D407 cells had a reduced viability on transfection with both plasmids. In vivo, subretinal injections of DNA-K8/JTS-1 complexes revealed reporter gene expression in choroidal and RPE cells of normal pink-eyed Royal College of Surgeons (RCS) rats. The validity of this transfection technique in terms of retinal cell survival in RCS rats was then examined by using pREP-hFGF2 plasmid, which encodes the human basic fibroblast growth factor isoforms (hFGF2). Subretinal injection of pREP-hFGF2-K8/JTS-1 complexes into 3-week-old dystrophic RCS rat eyes reveals a delayed photoreceptor cell degeneration 60 days postinjection. In this case, the average analyzed field points with delayed cell dystrophy represent 14 to 17% of the retinal surface as compared with 2.6 and 4% in pREP5beta and vehicle-injected eyes, respectively. Peptide-mediated in oculo transfection thus appears to be a promising technique for the treatment of retinal cell and photoreceptor degenerations.</abstract><cop>Larchmont, NY</cop><pub>Liebert</pub><pmid>10986560</pmid><doi>10.1089/10430340050129495</doi><tpages>16</tpages></addata></record> |
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| source | Mary Ann Liebert Online Subscription; MEDLINE |
| subjects | Animals Base Sequence beta-Galactosidase - genetics beta-Galactosidase - metabolism Biological and medical sciences Biotechnology Cell Survival - genetics Fibroblast Growth Factor 2 - genetics Fibroblast Growth Factor 2 - metabolism Fibroblast Growth Factor 2 - pharmacology Fundamental and applied biological sciences. Psychology Gene therapy Genetic Therapy - methods Green Fluorescent Proteins Health. Pharmaceutical industry Humans Industrial applications and implications. Economical aspects Luminescent Proteins - genetics Luminescent Proteins - metabolism Microinjections Molecular Sequence Data Photoreceptor Cells, Vertebrate - cytology Photoreceptor Cells, Vertebrate - pathology Photoreceptor Cells, Vertebrate - physiology Pigment Epithelium of Eye - cytology Plasmids - genetics Rats Retinal Degeneration - genetics Transfection - methods |
| title | Ocular cell transfection with the human basic fibroblast growth factor gene delays photoreceptor cell degeneration in RCS rats |
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