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@ARTICLE{Vlkner:155698,
      author       = {Völkner, Manuela and Pavlou, Marina and Büning, Hildegard
                      and Michalakis, Stylianos and Karl, Mike Oliver},
      title        = {{O}ptimized {A}deno-{A}ssociated {V}irus {V}ectors for
                      {E}fficient {T}ransduction of {H}uman {R}etinal
                      {O}rganoids.},
      journal      = {Human gene therapy},
      volume       = {32},
      number       = {13-14},
      issn         = {1557-7422},
      address      = {New York, NY},
      publisher    = {Liebert},
      reportid     = {DZNE-2021-00866},
      pages        = {694 - 706},
      year         = {2021},
      abstract     = {The most widely used vectors for gene delivery in the
                      retina are recombinant adeno-associated virus (rAAV)
                      vectors. They have proven to be safe and effective in
                      retinal gene therapy studies aimed to treat inherited
                      retinal dystrophies, although with various limitations in
                      transduction efficiency. Novel variants with modified capsid
                      sequences have been engineered to improve transduction and
                      overcome limitations of naturally occurring variants.
                      Although preclinical evaluation of rAAV vectors based on
                      such novel capsids is mostly done in animal models, the use
                      of human induced pluripotent stem cell (hiPSC)-derived
                      organoids offers an accessible and abundant human testing
                      platform for rAAV evaluation. In this study, we tested the
                      novel capsids, AAV9.GL and AAV9.NN, for their tropism and
                      transduction efficiency in hiPSC-derived human retinal
                      organoids (HROs) with all major neuronal and glial cell
                      types in a laminated structure. These variants are based on
                      the AAV9 capsid and were engineered to display specific
                      surface-exposed peptide sequences, previously shown to
                      improve the retinal transduction properties in the context
                      of AAV2. To this end, HROs were transduced with increasing
                      concentrations of rAAV9, rAAV9.GL, or rAAV9.NN carrying a
                      self-complementary genome with a cytomegalovirus-enhanced
                      green fluorescent protein (eGFP) cassette and were monitored
                      for eGFP expression. The rAAV vectors transduced HROs in a
                      dose-dependent manner, with rAAV9.NN achieving the highest
                      efficiency and fastest onset kinetics, leading to detectable
                      eGFP signals in photoreceptors, some interneurons, and
                      Müller glia already at 2 days post-transduction. The
                      potency-enhancing effect of the NN peptide insert was
                      replicated when using the corresponding AAV2-based version
                      (rAAV2.NN). Taken together, we report the application of an
                      HRO system for screening novel AAV vectors and introduce
                      novel vector candidates with enhanced transduction
                      efficiency for human retinal cells.},
      keywords     = {Animals / Dependovirus: genetics / Gene Transfer Techniques
                      / Genetic Vectors: genetics / Green Fluorescent Proteins:
                      genetics / Humans / Induced Pluripotent Stem Cells /
                      Organoids / Retina / Transduction, Genetic /
                      adeno-associated virus (Other) / engineered AAV vector
                      (Other) / human retinal organoids (Other)},
      cin          = {AG Karl},
      ddc          = {610},
      cid          = {I:(DE-2719)1710004},
      pnm          = {352 - Disease Mechanisms (POF4-352)},
      pid          = {G:(DE-HGF)POF4-352},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:33752467},
      doi          = {10.1089/hum.2020.321},
      url          = {https://pub.dzne.de/record/155698},
}