title: Twin disulfides as opportunity for improving stability and transfection efficiency of oligoaminoethane polyplexes.
abstract: The synthesis of precise gene delivery vehicles by solid-supported chemistry is an effective way to establish structure-activity relationships and optimize existing transfection carriers. Sequence-defined cationic oligomers with different topologies were modified with twin disulfide-forming cysteine-arginine-cysteine (CRC) motifs. The influence of this motif versus single disulfide on the biophysical properties and biological performance of polyplexes was investigated, with pDNA and siRNA as nucleic acid cargoes. Clear differences between structures with isolated cysteines and CRC motifs were observed with respect to properties like nucleic acid binding, serum stability, response to reducing agents, and gene transfer/silencing. The main observed effect of the CRC motif was to increase polyplex stability. The consequences for nucleic acid delivery were less predictable and depended on oligomer topology. For some oligomers intrinsically forming stable polyplexes (i.e., already in the absence of CRC motif), this further stabilization resulted in a reduction or even loss in transfection efficiency. For PEGylated and targeted oligomers with intrinsically less stable polyplex structures, this modification led to a significant enhancement in transfection efficiency.
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Philipp M
authors.initials: PM
authors.lastname: Klein
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Müller
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Gutmann
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Kos
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Ana
authors.initials: A
authors.lastname: Krhac Levacic
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Edinger
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Miriam
authors.initials: M
authors.lastname: Höhn
authors.affiliation: Swiss Federal Institute of Technology Zurich (ETHZ), Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, Vladimir Prelog Weg 3, CH-8093 Zurich, Switzerland.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Leroux
authors.affiliation: Institut National de la Recherche Scientifique (INRS), EMT Research Centre, 1650 boul. Lionel-Boulet, Varennes, Quebec J3X 1S2, Canada.
authors.forename: Marc A
authors.initials: MA
authors.lastname: Gauthier
authors.affiliation: Ludwig-Maximilians-University (LMU) Munich, Pharmaceutical Biotechnology, Department of Pharmacy, Butenandtstrasse 5-13, D-81377 Munich, Germany; Nanosystems Initiative Munich, Schellingstraße 4, D-80799 Munich, Germany. Electronic address: ernst.wagner@cup.uni-muenchen.de.
authors.forename: Ernst
authors.initials: E
authors.lastname: Wagner
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001120:Arginine; D001692:Biological Transport; D002460:Cell Line; D003545:Cysteine; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D051379:Mice; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D011073:Polyamines / Q000737:chemistry / Q000378:metabolism*; D000071228:Polyelectrolytes; D011092:Polyethylene Glycols / Q000737:chemistry; D055550:Protein Stability; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014162:Transfection / Q000379:methods*
chemical_list: D001120:Arginine; D003545:Cysteine; D004220:Disulfides; D011073:Polyamines; D000071228:Polyelectrolytes; D011092:Polyethylene Glycols; D034741:RNA, Small Interfering
keywords: Bio-reduction; Nucleic acid delivery; Polyplex stabilization; Twin disulfide; pDNA; siRNA