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@ARTICLE{Rogler:284075,
author = {Rogler, Teresa S and Salbaum, Katja A and Brinkop, Achim T
and Sonntag, Selina M and James, Rebecca and Shelton, Elijah
R and Thielen, Alina and Rose, Roland and Babutzka, Sabrina
and Klopstock, Thomas and Michalakis, Stylianos and Serwane,
Friedhelm},
title = {3{D} {Q}uantification of {V}iral {T}ransduction
{E}fficiency in {L}iving {H}uman {R}etinal {O}rganoids.},
journal = {Small Methods},
volume = {10},
number = {2},
issn = {2366-9608},
address = {Weinheim},
publisher = {WILEY-VCH Verlag GmbH $\&$ Co. KGaA},
reportid = {DZNE-2026-00083},
pages = {2401050},
year = {2025},
abstract = {The development of therapeutics builds on testing their
efficiency in vitro. To optimize gene therapies, for
example, fluorescent reporters expressed by treated cells
are typically utilized as readouts. Traditionally, their
global fluorescence signal has been used as an estimate of
transduction efficiency. However, analysis in individual
cells within a living 3D tissue remains a challenge. Readout
on a single-cell level can be realized via
fluorescence-based flow cytometry at the cost of tissue
dissociation and loss of spatial information. Complementary,
spatial information is accessible via immunofluorescence of
fixed samples. Both approaches impede time-dependent studies
on the delivery of the vector to the cells. Here,
quantitative 3D characterization of viral transduction
efficiencies in living retinal organoids is introduced. The
approach combines quantification of gene delivery efficiency
in space and time, leveraging human retinal organoids,
engineered adeno-associated virus (AAV) vectors, confocal
live imaging, and deep learning-based image segmentation.
The integration of these tools in an organoid imaging and
analysis pipeline allows quantitative testing of future
treatments and other gene delivery methods. It has the
potential to guide the development of therapies in
biomedical applications.},
keywords = {3D segmentation (Other) / adeno‐associated virus (Other)
/ gene therapy (Other) / retinal organoids (Other) /
transduction efficiency (Other)},
cin = {Clinical Research (Munich)},
ddc = {620},
cid = {I:(DE-2719)1111015},
pnm = {353 - Clinical and Health Care Research (POF4-353)},
pid = {G:(DE-HGF)POF4-353},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40509616},
doi = {10.1002/smtd.202401050},
url = {https://pub.dzne.de/record/284075},
}