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@ARTICLE{Cremer:829380,
author = {Cremer, Christian and Braun, Hanna and Mladenov, Radoslav
and Schenke, Lea and Cong, Xiaojing and Jost, Edgar and
Brümmendorf, Tim H. and Fischer, Rainer and Carloni, Paolo
and Barth, Stefan and Nachreiner, Thomas},
title = {{N}ovel angiogenin mutants with increased cytotoxicity
enhance the depletion of pro-inflammatory macrophages and
leukemia cells ex vivo},
journal = {Cancer immunology immunotherapy},
volume = {64},
number = {12},
issn = {1432-0851},
address = {Berlin},
publisher = {Springer},
reportid = {FZJ-2017-03093},
pages = {1575 - 1586},
year = {2015},
abstract = {Immunotoxins are fusion proteins that combine a targeting
component such as an antibody fragment or ligand with a
cytotoxic effector component that induces apoptosis in
specific cell populations displaying the corresponding
antigen or receptor. Human cytolytic fusion proteins (hCFPs)
are less immunogenic than conventional immunotoxins because
they contain human pro-apoptotic enzymes as effectors.
However, one drawback of hCFPs is that target cells can
protect themselves by expressing endogenous inhibitor
proteins. Inhibitor-resistant enzyme mutants that maintain
their cytotoxic activity are therefore promising effector
domain candidates. We recently developed potent variants of
the human ribonuclease angiogenin (Ang) that were either
more active than the wild-type enzyme or less susceptible to
inhibition because of their lower affinity for the
ribonuclease inhibitor RNH1. However, combining the
mutations was unsuccessful because although the enzyme
retained its higher activity, its susceptibility to RNH1
reverted to wild-type levels. We therefore used molecular
dynamic simulations to determine, at the atomic level, why
the affinity for RNH1 reverted, and we developed strategies
based on the introduction of further mutations to once again
reduce the affinity of Ang for RNH1 while retaining its
enhanced activity. We were able to generate a novel Ang
variant with remarkable in vitro cytotoxicity against HL-60
cells and pro-inflammatory macrophages. We also demonstrated
the pro-apoptotic potential of Ang-based hCFPs on cells
freshly isolated from leukemia patients.},
cin = {GRS Jülich ; German Research School for Simulation
Sciences / IAS-5},
ddc = {610},
cid = {I:(DE-Juel1)GRS-20100316 / I:(DE-Juel1)IAS-5-20120330},
pnm = {899 - ohne Topic (POF3-899)},
pid = {G:(DE-HGF)POF3-899},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000365176600008},
pubmed = {pmid:26472728},
doi = {10.1007/s00262-015-1763-8},
url = {https://juser.fz-juelich.de/record/829380},
}