September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
rAAV8 biodistribution and shedding after subretinal injection in non-human primates
Author Affiliations & Notes
  • Immanuel Philipp Seitz
    University Eye Hospital Tübingen, Tübingen, Germany
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • M. Dominik Fischer
    University Eye Hospital Tübingen, Tübingen, Germany
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Stylianos Michalakis
    Center for Integrated Protein Science Munich (CIPSM), Departement of Pharmacology, Ludwig-Maximilians-Universität München, Munich, Germany
  • Barbara Wilhelm
    STZ eyetrial, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Nadine Kahle
    STZ eyetrial, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Eberhart Zrenner
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Marius Ueffing
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Karl Ulrich Bartz-Schmidt
    University Eye Hospital Tübingen, Tübingen, Germany
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Martin Biel
    Center for Integrated Protein Science Munich (CIPSM), Departement of Pharmacology, Ludwig-Maximilians-Universität München, Munich, Germany
  • Bernd Wissinger
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Tobias Peters
    Institute for Ophthalmic Research, University of Tübingen, Centre for Ophthalmology, Tübingen, Germany
  • Footnotes
    Commercial Relationships   Immanuel Philipp Seitz, None; M. Dominik Fischer, None; Stylianos Michalakis, None; Barbara Wilhelm, None; Nadine Kahle, None; Eberhart Zrenner, None; Marius Ueffing, None; Karl Ulrich Bartz-Schmidt, None; Martin Biel, None; Bernd Wissinger, None; Tobias Peters, None
  • Footnotes
    Support  Tistou & Charlotte Kerstan Stiftung
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4025. doi:
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      Immanuel Philipp Seitz, M. Dominik Fischer, Stylianos Michalakis, Barbara Wilhelm, Nadine Kahle, Eberhart Zrenner, Marius Ueffing, Karl Ulrich Bartz-Schmidt, Martin Biel, Bernd Wissinger, Tobias Peters; rAAV8 biodistribution and shedding after subretinal injection in non-human primates. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4025.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : The aim of this study was to analyse virus distribution and shedding after a single subretinal administration of clinical grade recombinant adeno-associated virus (rAAV8) in non-human primates. This is important for environmental risk assessment in retinal gene therapy trials.

Methods : 18 non-human primates (Macacca fascicularis) underwent 23G pars plana vitrectomy and subretinal injection in three cohorts (high dose: 1x1012 vector genomes [vg], low dose: 1x1011 vg, or vehicle only). Four additional animals received intravitreal injections to mimic via falsa biodistribution. Tissues samples were harvested at necropsy (day 91) from the treated eye, draining lymph nodes, salivary gland and spleen, optic nerve, brain and spinal cord, heart, lung, liver, adrenal glands and gonads. Blood, urine, lacrimal and nasal swabs were harvested from each animal prior to dosing and 1, 2 and 3 days and 1, 4 and 13 weeks after application of the vector for DNA extraction and quantification of vector genomes by qPCR.

Results : Dose dependent rAAV8 DNA was found in the treated retina. Persistence of rAAV8 in other tissues was most notable after intravitreal injection (via falsa control). Shedding was found in all bio fluids. The highest concentrations were found in lacrimal fluid of the high dose group. DNA was not detected in the germ line tissues and apart from sporadic signals detected in a small number of animals in the liver and lung, all remaining tissues were negative. Concerning the biofluids at day 31 only blood samples showed remaining virus DNA.

Conclusions : These data are relevant for clinical retinal gene therapy trials, where trial subjects, investigators and regulators alike are interested to identify environmental risks associated with application of genetically modified organisms. While shedding into biofluids seems to occur in a dose dependent manner, transduction of off-target organs seems minimal.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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