Immanuel P Seitz1,2, Stylianos Michalakis3, Barbara Wilhelm4, Felix F Reichel1,2, G Alex Ochakovski1,2, Eberhart Zrenner2, Marius Ueffing2, Martin Biel3, Bernd Wissinger2, Karl U Bartz-Schmidt1, Tobias Peters4, M Dominik Fischer1,2,4,5. 1. University Eye Hospital, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany. 2. Institute for Ophthalmic Research, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany. 3. Center for Integrated Protein Science Munich (CIPSM) at the Department of Pharmacy-Center for Drug Research, Ludwig-Maximilians-Universität München, Munich, Germany. 4. STZ Eyetrial at the Centre for Ophthalmology, University of Tübingen, Tübingen, Germany. 5. Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, United Kingdom.
Abstract
Purpose: To investigate shedding and biodistribution characteristics of recombinant adeno-associated virus serotype 8 (rAAV8) after single-dose subretinal or intravitreal injection in nonhuman primates (NHP, Macaca fascicularis) as a surrogate for environmental hazard and patient safety. Methods: In a study for regulatory submission, 22 NHP were divided into four cohorts receiving either single subretinal injections of vehicle or clinical grade rAAV8 (1 × 1011 or 1 × 1012 vector genomes [vg]) versus single intravitreal application of 1 × 1012 vg. Viral shedding and biodistribution were monitored in biofluids for up to 91 days, followed by necropsy and tissue harvesting of all major organs, the visual pathway, and lymphatic tissue. Quantification of vector genomes was done by quantitative (q)PCR. Results: Shedding occurred in a dose-dependent manner in all biofluids and persisted for a maximum of 7 days. Intravitreal delivery led to increased and persistent (up to 13 weeks) distribution of vector genomes in blood and draining lymphatic tissue, increased off-target deposition, and inefficient gene transfer to the retina. No vector targeting of the germ line was observed in any cohort. Conclusions: These data illustrate that subretinal application of rAAV8 leads to a more favorable biodistribution profile compared to intravitreal injections. Extraocular biodistribution is limited after subretinal delivery, while intravitreal injection leads to both greater and more persistent systemic exposure, evident in blood and lymphatic tissues. With the knowledge on the dynamics of shedding in a setting mimicking clinical application, guidelines can be developed to refine clinical trial protocols to reduce the risk for trial subjects and their environment.
Purpose: To investigate shedding and biodistribution characteristics of recombinant adeno-associated virus serotype 8 (rAAV8) after single-dose subretinal or intravitreal injection in nonhuman primates (NHP, Macaca fascicularis) as a surrogate for environmental hazard and patient safety. Methods: In a study for regulatory submission, 22 NHP were divided into four cohorts receiving either single subretinal injections of vehicle or clinical grade rAAV8 (1 × 1011 or 1 × 1012 vector genomes [vg]) versus single intravitreal application of 1 × 1012 vg. Viral shedding and biodistribution were monitored in biofluids for up to 91 days, followed by necropsy and tissue harvesting of all major organs, the visual pathway, and lymphatic tissue. Quantification of vector genomes was done by quantitative (q)PCR. Results: Shedding occurred in a dose-dependent manner in all biofluids and persisted for a maximum of 7 days. Intravitreal delivery led to increased and persistent (up to 13 weeks) distribution of vector genomes in blood and draining lymphatic tissue, increased off-target deposition, and inefficient gene transfer to the retina. No vector targeting of the germ line was observed in any cohort. Conclusions: These data illustrate that subretinal application of rAAV8 leads to a more favorable biodistribution profile compared to intravitreal injections. Extraocular biodistribution is limited after subretinal delivery, while intravitreal injection leads to both greater and more persistent systemic exposure, evident in blood and lymphatic tissues. With the knowledge on the dynamics of shedding in a setting mimicking clinical application, guidelines can be developed to refine clinical trial protocols to reduce the risk for trial subjects and their environment.
Authors: Alejandra Bosco; Sarah R Anderson; Kevin T Breen; Cesar O Romero; Michael R Steele; Vince A Chiodo; Sanford L Boye; William W Hauswirth; Stephen Tomlinson; Monica L Vetter Journal: Mol Ther Date: 2018-08-24 Impact factor: 11.454
Authors: Catherine Cukras; Henry E Wiley; Brett G Jeffrey; H Nida Sen; Amy Turriff; Yong Zeng; Camasamudram Vijayasarathy; Dario Marangoni; Lucia Ziccardi; Sten Kjellstrom; Tae Kwon Park; Suja Hiriyanna; J Fraser Wright; Peter Colosi; Zhijian Wu; Ronald A Bush; Lisa L Wei; Paul A Sieving Journal: Mol Ther Date: 2018-07-07 Impact factor: 11.454
Authors: Douglas J Ballon; Jonathan B Rosenberg; Edward K Fung; Anastasia Nikolopoulou; Paresh Kothari; Bishnu P De; Bin He; Alvin Chen; Linda A Heier; Dolan Sondhi; Stephen M Kaminsky; Paul David Mozley; John W Babich; Ronald G Crystal Journal: Hum Gene Ther Date: 2020-12 Impact factor: 5.695
Authors: Alaknanda Mishra; Camasamudram Vijayasarathy; Catherine A Cukras; Henry E Wiley; H Nida Sen; Yong Zeng; Lisa L Wei; Paul A Sieving Journal: Mol Ther Date: 2021-02-15 Impact factor: 12.910