TY - JOUR
T1 - Stable retinal gene expression in nonhuman primates via subretinal injection of SIVagm-based lentiviral vectors
AU - Ikeda, Yasuhiro
AU - Yonemitsu, Yoshikazu
AU - Miyazaki, Masanori
AU - Kohno, Ri Ichiro
AU - Murakami, Yusuke
AU - Murata, Toshinori
AU - Tabata, Toshiaki
AU - Ueda, Yasuji
AU - Ono, Fumiko
AU - Suzuki, Toshimichi
AU - Ageyama, Naohide
AU - Terao, Keiji
AU - Hasegawa, Mamoru
AU - Sueishi, Katsuo
AU - Ishibashi, Tatsuro
PY - 2009/6/1
Y1 - 2009/6/1
N2 - Gene therapy may hold promise as a therapeutic approach for the treatment of intractable ocular diseases, including retinitis pigmentosa (RP). Gene transfer vectors that are able to show long-lasting transgene expression in vivo are highly desirable to treat RP; however, there is a dearth of information regarding long-term transgene expression in the eyes of large animals. We previously reported that the simian immunodeficiency virus from African green monkeys (SIVagm)-based lentiviral vector showed efficient, stable, and safe retinal gene transfer, resulting in significant prevention of retinal degeneration by gene transfer of a neurotrophic factor, human pigment epithelium-derived factor (hPEDF), in rodents. Before applying this strategy in a clinical setting, we here assessed the long-lasting transgene expression of our third-generation SIVagm-based lentiviral vectors in the retinal tissue of nonhuman primates. Approximately 20-50μl of SIV-EGFP (enhanced green fluorescent protein) or SIV-hPEDF was injected into the subretinal space via a glass capillary tube. To detect EGFP expression in the retina, we used a fluorescence fundus camera at various time points after gene transfer. Human PEDF expression was assessed by immunohistochemical analysis, Western blot assay, and enzyme-linked immunosorbent assay. The retinas demonstrated frequent EGFP expression that was preserved for at least 4 years without significant decline. The expression of hPEDF was stable, and occurred mainly in the retinal pigment epithelium. The secreted protein was detected in vitreous and aqueous humor. We thus propose that SIVagm-mediated stable gene transfer might be significantly useful for ocular gene transfer in a clinical setting.
AB - Gene therapy may hold promise as a therapeutic approach for the treatment of intractable ocular diseases, including retinitis pigmentosa (RP). Gene transfer vectors that are able to show long-lasting transgene expression in vivo are highly desirable to treat RP; however, there is a dearth of information regarding long-term transgene expression in the eyes of large animals. We previously reported that the simian immunodeficiency virus from African green monkeys (SIVagm)-based lentiviral vector showed efficient, stable, and safe retinal gene transfer, resulting in significant prevention of retinal degeneration by gene transfer of a neurotrophic factor, human pigment epithelium-derived factor (hPEDF), in rodents. Before applying this strategy in a clinical setting, we here assessed the long-lasting transgene expression of our third-generation SIVagm-based lentiviral vectors in the retinal tissue of nonhuman primates. Approximately 20-50μl of SIV-EGFP (enhanced green fluorescent protein) or SIV-hPEDF was injected into the subretinal space via a glass capillary tube. To detect EGFP expression in the retina, we used a fluorescence fundus camera at various time points after gene transfer. Human PEDF expression was assessed by immunohistochemical analysis, Western blot assay, and enzyme-linked immunosorbent assay. The retinas demonstrated frequent EGFP expression that was preserved for at least 4 years without significant decline. The expression of hPEDF was stable, and occurred mainly in the retinal pigment epithelium. The secreted protein was detected in vitreous and aqueous humor. We thus propose that SIVagm-mediated stable gene transfer might be significantly useful for ocular gene transfer in a clinical setting.
UR - http://www.scopus.com/inward/record.url?scp=68849083844&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=68849083844&partnerID=8YFLogxK
U2 - 10.1089/hum.2009.009
DO - 10.1089/hum.2009.009
M3 - Article
C2 - 19257835
AN - SCOPUS:68849083844
VL - 20
SP - 573
EP - 579
JO - Human Gene Therapy
JF - Human Gene Therapy
SN - 1043-0342
IS - 6
ER -