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@ARTICLE{Altai:299009,
author = {M. Altai and Á. Nagy and P. Granit and W. Zedan and M.
Cerezo-Magaña and J. Park and K. Lückerath$^*$ and S.
Geres and M. Sydoff and D. L. J. Thorek and K. Westerlund
and D. Ulmert and A. E. Karlström},
title = {{O}ptimizing peptide nucleic acid-based pretargeting for
enhanced targeted radionuclide therapy.},
journal = {Journal of controlled release},
volume = {381},
issn = {0168-3659},
address = {New York, NY [u.a.]},
publisher = {Elsevier},
reportid = {DKFZ-2025-00419},
pages = {113551},
year = {2025},
note = {Volume 381, 10 May 2025, 113551},
abstract = {Radiolabeled targeting agents have emerged as valuable
tools for the treatment of disseminated cancer. Monoclonal
antibodies (mAbs) are widely employed as carriers for
diagnostic and therapeutic radionuclides due to their
exceptional specificity and affinity. However, their
prolonged circulatory half-life can diminish diagnostic
efficacy and increase radiation exposure to non-target
tissues in therapeutic applications, resulting in
dose-limiting toxicities. To overcome this limitation,
pretargeting technologies emerge as promising strategies to
enhance tumor-to-background ratio and reduce radiation
exposure of healthy tissues. Our previous work introduced a
pretargeting concept leveraging the specific interaction
between two peptide nucleic acid (PNA) probes, HP1 and HP2,
as the recognition mechanism. This early iteration of the
PNA-based concept showed limited efficacy when used with
mAb-based vectors. To improve its performance, we
re-engineered the primary and secondary targeting agents by
incorporating newly designed PNA-probes. As the primary
targeting agent, we functionalized trastuzumab (T), a
well-characterized human epidermal growth factor receptor 2
(HER2)-targeting IgG1 mAb, with a 9-mer PNA probe (HP9).
Both FcIII-based covalent UV-light crosslinking and
enzyme-mediated glyco-engineering click-chemistry methods
were applied to generate trastuzumab-PNA conjugates
T-FcIII-HP9 and T-gly-HP9, respectively. As a
radionuclide-carrying secondary agent, we utilized a 9-mer
complementary PNA probe, HP16, which forms a stable duplex
with HP9 as well as displaying favorable in vivo kinetics.
Biacore and flow cytometry assessment of the HP9-conjugated
trastuzumab agents demonstrated retained HER2-binding
properties. The secondary HP16 probe, labeled with either a
dye or a radionuclide, showed cell surface accumulation
contingent on the presence of HP9 on the primary
HER2-targeting agents. In vivo, T-gly-HP9 exhibited
significantly longer blood circulation half-life and
superior tumor uptake compared to T-FcIII-HP9. Further,
therapeutic dosing with [177Lu]-HP16 of trastuzumab-HP9
pretargeted HER2+ tumor models resulted in significantly
delayed disease progression and extended survival compared
to untreated subjects. Furthermore, pretargeted [177Lu]-HP16
exhibited comparable efficacy to [177Lu]-trastuzumab in both
delaying disease progression and prolonging survival. In
conclusion, the optimization of our PNA-based pretargeting
system has resulted in exceptional in vivo targeting
characteristics and therapeutic efficacy, validating the
potential of this novel approach.},
keywords = {Antibody (Other) / Peptide nucleic acid (Other) /
Pretargeting (Other) / Radiotherapy (Other) / Site-specific
labeling (Other)},
cin = {ED01},
ddc = {610},
cid = {I:(DE-He78)ED01-20160331},
pnm = {899 - ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:39986477},
doi = {DOI:10.1016/j.jconrel.2025.02.047},
url = {https://inrepo02.dkfz.de/record/299009},
}