Prolonged transgene expression with lentiviral vectors in the aqueous humor outflow pathway of nonhuman primates.
We injected lentiviral vectors into the eyes of live nonhuman primates to assess potential for glaucoma gene therapy. Anterior chambers of five cynomolgus monkeys were injected with green fluorescent protein (GFP)-encoding feline immunodeficiency viral vectors. The monkeys were monitored for in vivo transgene expression and clinical parameters. Their eyes were harvested 2-15 months postinjection for tissue analyses. All seven eyes injected with 1.0-2.0 x 10(8) transducing units (TU) showed substantial GFP fluorescence in the trabecular meshwork (TM), which was observable even by goniophotographic monitoring for up to 15 months. Only the lowest dose (0.03 x 10(8) TU) failed to result in TM fluorescence detectable in vivo, and five of the eight vector-injected eyes continued to display substantial GFP expression when enucleated eyes were examined at 2, 7, or 15 months postinjection. Some transduced cells were also detected in the iris and ciliary body. Mild, transient postinjection inflammatory responses exceeding that induced by a control saline injection were observed, but vectors did not raise intraocular pressure and were well tolerated. The results demonstrate the first lentiviral vector transduction of the nonhuman primate aqueous humor outflow pathway and support application of the system to human glaucoma gene therapy.
Barraza
Román A
RA
Department of Molecular Medicine, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA.
Rasmussen
Carol A
CA
Loewen
Nils
N
Cameron
J Douglas
JD
Gabelt
B'Ann T
BT
Teo
Wu-Lin
WL
Kaufman
Paul L
PL
Poeschla
Eric M
EM
D000818:Animals; D001082:Aqueous Humor / Q000166:cytology / Q000378:metabolism*; D002924:Ciliary Body / Q000166:cytology / Q000378:metabolism; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D005902:Glaucoma, Open-Angle / Q000628:therapy; D049452:Green Fluorescent Proteins; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000235:genetics*; D007267:Injections; D007498:Iris / Q000166:cytology / Q000378:metabolism; D008252:Macaca fascicularis; D008297:Male; D011994:Recombinant Proteins / Q000096:biosynthesis; D013997:Time Factors; D014129:Trabecular Meshwork / Q000166:cytology / Q000378:metabolism; D019076:Transgenes / Q000502:physiology*
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins