Nanotechnology for Nucleic Acid Delivery: Methods and Protocols

299 Pages·2013·6.1 MB·English

by  Maria Dusinska

M M B ™
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hat fi eld, Hertfordshire, AL10 9AB, UK
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Nanotechnology for Nucleic
Acid Delivery
Methods and Protocols
Edited by
Manfred Ogris
Department of Pharmacy,
Ludwig-Maximilians-University Munich, Munich, Germany
David Oupicky
Department of Pharmaceutical Sciences,
Wayne State University, Detroit, MI, USA
Editors
Manfred Ogris David Oupicky
Department of Pharmacy Department of Pharmaceutical Sciences
Ludwig-Maximilians-University Munich Wayne State University
Munich, Germany Detroit, MI, USA
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-62703-139-4 ISBN 978-1-62703-140-0 (eBook)
DOI 10.1007/978-1-62703-140-0
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2012948771
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Preface
Nanotechnology and nucleic acid-based therapies are two emerging fi elds in science whose
combination has the potential to improve quality of life for patients suffering from various
diseases that can so far be treated only in an unsatisfactory way either due to excessive side
effects, such as in the case of malignant diseases, or due to the missing availability of thera-
peutically active compounds. Nucleic acids offer the potential for highly selective treatment
of such diseases, for example the specifi c expression of therapeutically active proteins with
gene vectors, or the highly specifi c modulation of gene expression with RNA interference.
Within gene therapy approaches, the DNA delivered should act in principle as a completely
inactive prodrug, which is activated only at the target site when being fi nally translated into
the therapeutically active protein. The same applies for approaches based on small interfer-
ing RNA (siRNA), where rather short pieces of RNA have to be delivered to the appropri-
ate site in the organism.
A key issue for successful nucleic acid therapies is the availability of a suitable delivery
system. Viral vectors, e.g., recombinant adenovirus or lentiviral vectors, are highly effi cient
on the cellular level after local delivery but have to be optimized for systemic gene delivery
approaches. In order to reduce residual biological risks of insertional mutagenesis,
in fl ammation, or other adverse reactions after virus injection, numerous synthetic nucleic
acid delivery devices have been developed within the last two decades. Here, the fi eld of
nanotechnology offers a multitude of possibilities to develop nanosized delivery vectors
tailor-made for various local and systemic approaches.
As for many novel fi elds in modern medicine, safety aspects are of great concern in the
development of nanocarriers for biomedical application. In Chapter 1 , Maria Dusinska and
colleagues report on safety aspects of engineered nanomaterials, including concerns related
to biodistribution and bioavailability and cellular aspects, as well as regulatory issues.
Chapters 2 and 3 by Moein Moghimi et al. describe the application of two assays to mea-
sure cellular toxicity of polycationic nucleic acid carriers, namely lactate dehydrogenase
release as a direct measure of cell membrane disruption, and the quantifi cation of caspase
activation, which leads to apoptosis. Nanoparticle properties are known to strongly infl uence
their in vivo behavior, like biodistribution or their interaction with the body defense system.
Immunological aspects of the in vivo use of nucleic acid delivery are addressed in Chapter
4 by Tatsuhiro Ishida with a method to measure the induction of IgM by polyethylenegly-
col, a polymer often used for surface coating of nanoparticles. In Chapter 5 , Jean-Luc Coll
and colleagues present a set of methods to determine the biodistribution of fl uorescently
labeled nucleic acid carriers in the living organism utilizing near infrared imaging.
Delivery systems based on lipids or cationic polymers are widely used for nucleic acids
forming the so-called lipoplexes or polyplexes, respectively. In Chapter 6 , Nathalie Mignet
et al. describe the synthesis of charge neutral, thiourea-based lipids and the formation of
liposomal nanoparticles with plasmid DNA. Antoine Kichler and colleagues in Chapter 7
give insights on the use of cationic peptides for nucleic acid delivery, and in Chapter 8 , Rödl
et al. report on the synthesis of conjugates of the polycation polyethylenimine and cell
v
vi Preface
targeting peptides for tumor cell-directed gene delivery with polyplexes. Polycation topol-
ogy is known to affect their toxicity and transfection activity. Several synthetic methods of
controlling topology of bioreducible polycations is by Ye-Zi You et al. in Chapter 9 . Nucleic
acid-based nanoparticles are often made fresh by mixing nucleic acid and carrier molecule.
Availability of stable formulations is crucial for preclinical and clinical applications. In
Chapter 10 , Julia Kasper and colleagues present a method for freeze-drying of polyplexes.
Angela Pannier and Tatjana Segura (Chapter 1 1 ) report a system based on polyplexes,
which are incorporated into a proteinous layer forming a hydrogel for surface-mediated
gene delivery. Miriam Breunig and colleagues in Chapter 1 2 describe the potential of layer-
by-layer fi lms of therapeutic nucleic acids deposited on gold nanoparticles.
Disassembly of nanoparticles and subsequent release of nucleic acids is an important
step for intracellular bioavailability of the delivered nucleic acids. In Chapter 1 3 , Mao and
colleagues describe an in situ atomic force microscopy method to study the disassembly of
polyplexes under various conditions in liquid.
Biological activity of nucleic acids in local or systemic administration can be improved
by combination with a range of physical methods. Niek Sanders et al. use focused ultra-
sound for site-directed disruption of gas-fi lled microbubbles, which enhance local gene
delivery (Chapter 14 ). Dialechti Vlaskou et al. describe the synthesis and application of
magnetic nanoparticles for DNA and RNA delivery, where targeting is achieved by applying
a magnetic fi eld (Chapter 1 5 ). Subsequent Chapter 1 6 by Yoshihisa Namiki describes the
synthesis of “lipomag,” a magnetic, lipidic DNA and RNA delivery system.
Chapters 1 7 – 19 present nanosized delivery systems and tissue-specifi c delivery strate-
gies. In Chapter 1 7 , Mark Zabel describes the synthesis and in vivo application of lipid- and
peptide-based nanoparticles for siRNA delivery into the brain, which readily cross the
blood–brain barrier after systemic injection. After systemic delivery, positively charged lipo-
plexes or polyplexes often end up in the fi rst vascular bed encountered, namely the lung
leading to transfection if lung tissue. Merkel and colleagues (Chapter 1 8 ) present a detailed
protocol to analyze cell types in lung transfected after transgene delivery with the help of
fl ow cytometry. Delivering nucleic acids to the liver can be achieved either by systemic
administration or by direct infusion into the portal vein. Mao and colleagues present an
alternative approach by transferring nanoparticulate systems via retrograde intrabiliary infu-
sion (Chapter 1 9 ).
Taken together, this book covers the area of nanoparticulate delivery of nucleic acids in
terms of biosafety, particle synthesis, and also its application in cell culture and after differ-
ent routes of administration in vivo.
Munich, Germany Manfred Ogris
Detroit, MI, USA David Oupicky
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Toxicological Aspects for Nanomaterial in Humans. . . . . . . . . . . . . . . . . . . . . 1
Maria Dusinska, Zuzana Magdolenova, and Lise Marie Fjellsbø
2 Lactate Dehydrogenase Assay for Assessment of Polycation Cytotoxicity . . . . . 13
Ladan Parhamifar, Helene Andersen, and S. Moein Moghimi
3 Combined Fluorimetric Caspase 3/7 Assay and Bradford Protein
Determination for Assessment of Polycation-Mediated Cytotoxicity . . . . . . . . 23
Anna K. Larsen, Arnaldur Hall, Henrik Lundsgart, and S. Moein Moghimi
4 Anti-PEG IgM Production via a PEGylated Nano-Carrier System
for Nucleic Acid Delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Masako Ichihara, Naoto Moriyoshi, Amr S. Abu Lila, Tatsuhiro Ishida,
and Hiroshi Kiwada
5 Near-Infrared Optical Imaging of Nucleic Acid Nanocarriers In Vivo . . . . . . . 49
Claire Rome, Julien Gravier, Marie Morille, Gilles Divita,
Anne-Laure Bolcato-Bellemin, Véronique Josserand, and Jean-Luc Coll
6 Lipids for Nucleic Acid Delivery: Synthesis and Particle Formation . . . . . . . . . 67
Michel Bessodes and Nathalie Mignet
7 Histidine-Rich Cationic Amphipathic Peptides for Plasmid DNA
and siRNA Delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Antoine Kichler, A. James Mason, Arnaud Marquette,
and Burkhard Bechinger
8 Synthesis of Polyethylenimine-Based Nanocarriers for Systemic Tumor
Targeting of Nucleic Acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Wolfgang Rödl, David Schaffert, Ernst Wagner, and Manfred Ogris
9 Synthesis of Bioreducible Polycations with Controlled Topologies. . . . . . . . . . 121
Ye-Zi You, Jun-Jie Yan, Zhi-Qiang Yu, and David Oupicky
10 Lyophilization of Synthetic Gene Carriers. . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Julia Christina Kasper, Sarah Küchler, and Wolfgang Friess
11 Surface-and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles. . . . . 149
Angela K. Pannier and Tatiana Segura
12 Layer-by-Layer Assembled Gold Nanoparticles for the Delivery
of Nucleic Acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
Eva-Christina Wurster, Asmaa Elbakry, Achim Göpferich,
and Miriam Breunig
vii
viii Contents
13 In Situ AFM Analysis Investigating Disassembly of DNA Nanoparticles
and Nano-Films . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Yi Zou, Lei Wan, Jenifer Blacklock, David Oupicky, and Guangzhao Mao
14 Enhancing Nucleic Acid Delivery with Ultrasound and Microbubbles. . . . . . . 195
Steven K. Cool, Bart Geers, Ine Lentacker, Stefaan C. De Smedt,
and Niek N. Sanders
15 Magnetic and Acoustically Active Microbubbles Loaded with Nucleic Acids
for Gene Delivery. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
Dialechti Vlaskou, Christian Plank, and Olga Mykhaylyk
16 Synthesis of Lipidic Magnetic Nanoparticles for Nucleic Acid Delivery . . . . . . 243
Yoshihisa Namiki
17 Lipopeptide Delivery of siRNA to the Central Nervous System. . . . . . . . . . . . 251
Mark D. Zabel
18 Flow Cytometry-Based Cell Type-Specifi c Assessment of Target Regulation
by Pulmonary siRNA Delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 263
Olivia M. Merkel, Leigh M. Marsh, Holger Garn, and Thomas Kissel
19 Liver-Targeted Gene Delivery Through Retrograde Intrabiliary Infusion. . . . . 275
Xuan Jiang, Yong Ren, John-Michael Williford, Zhiping Li,
and Hai-Quan Mao
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
Contributors
AMR S. ABU LILA (cid:129) Department of Pharmaceutics and Industrial Pharmacy,
Faculty of Pharmacy, Zagazig University, Zagazig , Egypt
HELENE ANDERSEN (cid:129) Centre for Pharmaceutical Nanotechnology , University of Copenha-
gen , Copenhagen Ø, Denmark
BURKHARD BECHINGER (cid:129) Institut de Chimie, Université de Strasbourg/CNRS UMR 7177,
Strasbourg , France
MICHEL BESSODES (cid:129) Unité de Pharmacologie Chimique et Génétique, CNRS, UMR 8151,
Paris , France; Inserm, U 1022, Paris , France; Faculté des Sciences Pharmaceutiques
et Biologiques, Université Paris Descartes, Paris , France; ENSCP , Paris , France
JENIFER BLACKLOCK (cid:129) Materials Reliability Division, National Institute of Standards
and Technology , Boulder , CO , USA
ANNE-LAURE BOLCATO-BELLEMIN (cid:129) Polyplus-transfection, Illkirch, France
MIRIAM BREUNIG (cid:129) Department of Pharmaceutical Technology , University of Regensburg,
Regensburg, Germany
JEAN-LUC COLL (cid:129) INSERM U823, Institut Albert Bonniot, Grenoble, France;
Université Joseph Fourier, Grenoble, France
STEVEN K. COOL (cid:129) Laboratory for Gene Therapy, Faculty of Veterinary Medicine,
Ghent University, Merelbeke, Belgium;
Ghent Research group on Nanomedicines, Laboratory of General Biochemistry
and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University,
Ghent , Belgium
STEFAAN C. DE SMEDT (cid:129) Ghent Research group on Nanomedicines, Laboratory of
General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences,
Ghent University, Ghent , Belgium
GILLES DIVITA (cid:129) CNRS UMR5237, Montpellier, France
MARIA DUSINSKA (cid:129) NILU, CEE, Health Effects Group, Kjeller , Norway
ASMAA ELBAKRY (cid:129) Department of Pharmaceutics, Faculty of Pharmacy,
Al-Azhar University, Cairo , Egypt
LISE MARIE FJELLSBØ (cid:129) NILU, CEE, Health Effects Group, Kjeller , Norway
WOLFGANG FRIESS (cid:129) Department of Pharmacy, Pharmaceutical Technology
and Biopharmaceutics, Ludwig-Maximilians-Universität, Munich, Germany
HOLGER GARN (cid:129) Institute of Laboratory Medicine and Pathobiochemistry,
Molecular Diagnostics, Philipps-Univeristät Marburg, Marburg , Germany;
Sterna biologicals GmbH & Co. KG, Marburg , Germany
BART GEERS (cid:129) Ghent Research group on Nanomedicines, Laboratory of General
Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences,
Ghent University, Ghent , Belgium
ACHIM GÖPFERICH (cid:129) Department of Pharmaceutical Technology , University of Regensburg,
Regensburg, Germany
ix