Polymeric Gene Delivery: Principles and Applications

by Amiji; Mansoor M.

• ISBN13: 9780849319341
• ISBN10: 084931934X
• Format: Hardcover
• Copyright: 2004-09-29
• Publisher: CRC Press

Summary

To treat disease or correct genetic disorders using gene therapy, the most suitable vehicle must be able to deliver genes to the appropriate tissues and cells in the body in a specific as well as safe and effective manner. While viruses are the most popular vehicles to date, their disadvantages include toxicity, limited size of genes they can carry, and limited scale of industrial production.Polymeric Gene Delivery: Principles and Applications is the first comprehensive book to specifically address polymeric gene delivery systems. Uniting the expertise of international academic and industrial scientists who are working in the area of polymeric vectors for gene delivery, it is written by prominent researchers directly involved in this field. The book is divided into five sections that deal with challenges and opportunities in gene delivery and the efficient delivery of genes into somatic cells using polymeric vectors. The authors discuss using biodegradable polymers, condensing and non-condensing polymeric systems, microspheres and nanospheres, and designing specialized delivery systems based on targeting strategies.Polymeric Gene Delivery: Principles and Applications accentuates the versatility of polymeric delivery systems, including the potential for biocompatibility, the ability to design their formulation and geometry for a specific purpose, and the ease of modification to the surface of polymeric carriers. This book is an up-to-date guide for researchers in the field and those interested in entering this dynamic field.

Table of Contents

Introduction	1	(4)
Robert Langer
Part I Gene Delivery: Challenges and Opportunities
Tissue- and Cell-Specific Targeting for the Delivery of Genetic Information	5	(24)
Randall J. Mrsny
Introduction	5	(1)
Background Issues	6	(2)
General Factors to Consider	8	(5)
Viral Systems	8	(2)
Nonviral Systems	10	(2)
Immune Concerns	12	(1)
Targeting Issues	12	(1)
Targeting Cells and Tissues	13	(1)
Topical Delivery Approaches	14	(1)
Local Injection	15	(1)
Vascular Targeting	16	(1)
Cell-Based Targeting	17	(2)
Conclusions	19	(10)
References	20	(9)
Biological Barriers to Gene Transfer	29	(14)
Yasufumi Kaneda
Introduction	29	(1)
Biological Barriers	30	(7)
Step 1: Reaching Target Cells	30	(1)
Recognition by Specific Target Tissues	30	(1)
Avoidance of Nonspecific Uptake	31	(1)
Resistance to Degradation in the Systemic Circulation	32	(1)
Step 2: Crossing Over the Cell Membrane	32	(1)
Step 3: Nuclear Targeting	33	(1)
Step 4: Regulation of Gene Expression	34	(1)
Stable Retention of a Transgene	35	(1)
Regulation of Transcription	36	(1)
Conclusion	37	(6)
References	37	(6)
Cellular Uptake and Trafficking	43	(20)
Sujatha Dokka
Yon Rojanasakul
Introduction	44	(1)
Viruses: Why Are They Such Efficient Delivery Systems?	44	(1)
Cellular Barriers to DNA Delivery	45	(7)
Stability in Extracellular Compartments	45	(1)
Association of DNA with the Cell Membrane	45	(1)
Cellular Internalization	46	(1)
Proton Sponge	46	(1)
pH-Sensitive Liposomes	46	(1)
Fusogenic Peptides	47	(1)
Cytosolic Transport of DNA	47	(1)
Metabolic Instability of DNA in the Cytosol	47	(1)
Nuclear Localization of Plasmid DNA	47	(1)
Nucleocytoplasmic Transport	48	(1)
Nuclear Import of DNA	49	(1)
Attempts to Increase the Efficiency of Nuclear DNA Uptake	49	(3)
Current Approaches for Enhancing Gene Delivery	52	(3)
Viral Vectors	52	(1)
Chemical/Synthetic Nonviral Vectors	53	(1)
Liposomes	53	(1)
DNA-Protein Conjugates	53	(1)
Physical Methods of DNA Delivery	54	(1)
Electroporation	54	(1)
Direct DNA Injection and Microinjection	54	(1)
Bombardment of DNA-Coated Particles	55	(1)
Other Physical Approaches	55	(1)
Conclusions	55	(8)
References	56	(7)
Pharmacokinetics of Polymer--Plasmid DNA Complex	63	(16)
Makiya Nishikawa
Yoshinobu Takakura
Mitsuru Hashida
Introduction	63	(1)
Pharmacokinetic Consideration of Macromolecules	64	(3)
Basic Pharmacokinetic Properties of Naked Plasmid DNA	67	(2)
Cellular Uptake Mechanism	67	(1)
Tissue Distribution after Intravenous Injection	68	(1)
Pharmacokinetics of Polymer--Plasmid DNA Complex	69	(1)
Cellular Uptake	69	(1)
Interaction with Biological Components	70	(1)
Tissue Distribution	70	(1)
Pharmacokinetics of Targeted Polymer--Plasmid DNA Complex	70	(3)
Fundamentals of Targeted Delivery	70	(1)
Sugar-Mediated Targeting	71	(2)
Conclusion	73	(6)
References	73	(6)
Part II Condensing Polymeric Systems
Non-Degradable Polymers
Poly(L-Lysine) and Copolymers for Gene Delivery	79	(18)
Minhyung Lee
Sung Wan Kim
Introduction	79	(1)
Characteristics of PLL as a Gene Carrier	80	(2)
PLL Conjugates	82	(9)
PLL Conjugates for Low Cytotoxicity	82	(1)
PLL--PEG Block Copolymer	82	(1)
PEG-Grafted PLL	83	(1)
PLL-Based Biodegradable Polymer	83	(1)
PLL Conjugates for Receptor-Mediated Endocytosis	84	(1)
PLL-Asialoorosomucoid	85	(1)
PLL-Transferrin	85	(1)
Galactose-PLL and Lactose-PLL	86	(1)
PLL-Antibody	86	(1)
PLL-Folate	87	(1)
The Terplex System	87	(1)
Artery-Wall-Binding Peptide PEG-Grafted-PLL	88	(1)
PLL Conjugates for Efficient Intracellular Trafficking	89	(1)
Histidylated PLL	89	(1)
Histidylated Oligolysines	90	(1)
Poly(histidine)-Grafted PLL	90	(1)
Fusogenic Peptides	90	(1)
PLL-NLS	90	(1)
Conclusions	91	(6)
References	91	(6)
Gene Delivery Using Polyethylenimine and Copolymers	97	(10)
Manfred Ogris
Introduction	97	(1)
Polyethylenimine: Chemistry and Derivatives	98	(1)
Coupling Strategies for PEI Conjugates	98	(1)
Protein-PEI and Peptide-PEI Conjugates	98	(1)
Biophysics of Nucleic Acid-PEI Interaction	98	(1)
PEI Polyplexes for Nucleic Acid Delivery	99	(1)
Ligand-PEI Conjugates for Gene Delivery	99	(1)
Intracellular Aspects of PEI-Mediated Gene Delivery	100	(1)
In Vivo Applications	101	(1)
Local Delivery	101	(1)
Systemic Application	102	(1)
Therapeutic Gene Delivery with PEI	102	(1)
Conclusions	103	(4)
References	103	(4)
Poly(2-(dimethylamino)ethyl methacrylate)-Based Polymers for the Delivery of Genes In Vitro and In Vivo	107	(26)
F.J. Verbaan
D.J.A. Crommelin
W.E. Hennink
G. Storm
Introduction	107	(1)
In Vitro Transfection Efficiency of pDMAEMA-Based Complexes: Critical Parameters	108	(8)
Physicochemical Properties	108	(2)
Design of pDMAEMA-Based Copolymers	110	(2)
Cellular Interactions of pDMAEMA-Based Polyplexes	112	(2)
Targeting of pDMAEMA-Based Polyplexes	114	(2)
Pharmaceutical Aspects of pDMAEMA-Based Complexes	116	(9)
Preparation of Highly Concentrated pDMAEMA-Polyplexes	116	(2)
Structural Properties of Plasmid DNA	118	(2)
Stability of pDMAEMA-Polyplexes	120	(1)
Freeze-Drying and Freeze-Thawing of pDMAEMA-Polyplexes	120	(2)
Long-Term Stability	122	(3)
In Vivo Delivery of pDMAEMA-Based Complexes to Tumors	125	(8)
Intraperitoneal Administration	125	(2)
Intravenous Administration	127	(3)
References	130	(3)
Cationic Dendrimers as Gene Transfection Vectors: Dendri-Poly(amidoamines) and Dendri-Poly(propylenimines)	133	(26)
Lori A. Kubasiak
Donald A. Tomalia
Introduction	134	(3)
Dendritic Polymers: A Major New Architectural Class	134	(1)
Dendrons and Dendrimers	135	(2)
Poly(amidoamine) (PAMAM) Dendrimers	137	(13)
Synthesis	137	(1)
Structure	138	(1)
Dendrimer Shape Changes: Nanoscale Container and Scaffolding Properties	138	(1)
Nanoscale Container and Scaffolding Properties	139	(1)
Nanoscale Dimensions and Shapes Mimicking Proteins	139	(2)
Cationic (PAMAM) Dendrimer/DNA Interactions	141	(1)
Cellular Entry	142	(1)
In Vitro Transfection	142	(1)
Plasmids	142	(2)
Oligonucleotides	144	(1)
Enhanced Dendrimer-Mediated Transfection	145	(1)
In Vitro Toxicity	146	(1)
Ex Vivo Gene Delivery	147	(1)
In Vivo Gene Delivery	148	(1)
Biodistribution	148	(1)
In Vivo Toxicity	149	(1)
Poly(propylenimine) (PPI) Dendrimers	150	(2)
Synthesis	150	(1)
Structure	150	(1)
DNA--Dendrimer Interactions	151	(1)
In Vitro Transfection	151	(1)
In Vitro Toxicity	151	(1)
Targeted Delivery	152	(1)
Conclusion	152	(7)
References	153	(6)
Poly(ethylene glycol)-Conjugated Cationic Dendrimers	159	(16)
Joon Sig Choi
Tae-il Kim
Jong-sang Park
Introduction	159	(1)
Synthesis and Characterization of the Hybrid Block Copolymers	160	(4)
MPEG-PLLD and PLLD-PEG-PLLD (Figures 10.2A and 10.2B)	160	(1)
Liquid Phase Peptide Synthesis	160	(1)
1H NMR and MALDI-TOF MS	161	(1)
MPEG-PAMAM and PAMAM-PEG-PAMAM (Figures 10.2C and 10.2D)	161	(1)
Synthesis of the Block Copolymers	161	(2)
1H NMR and MALDI-TOF MS	163	(1)
MPEG-PEI and PEG-PEI (Figures 10.2E and 10.2F)	163	(1)
PEG Activation and Conjugation	163	(1)
1H NMR	163	(1)
Self-Assembly of Block Copolymers with Plasmid DNA	164	(5)
Electrophoretic Mobility Shift Assay	164	(1)
Atomic Force Microscopy (AFM)	164	(3)
DNase I Protection Assay	167	(1)
Zeta Potential Measurements	168	(1)
Evaluation of Cytotoxicity In Vitro	169	(1)
Application to In Vitro Transfection	169	(3)
Conclusion	172	(3)
Acknowledgments	172	(1)
References	172	(3)
Water Soluble Lipopolymers for Gene Delivery	175	(12)
Ram I. Mahato
Sung Wan Kim
Introduction	175	(1)
Basic Components Of Cationic Lipids	175	(1)
Why Water-Soluble Lipopolymers?	176	(1)
Synthesis and Characterization of Water-Soluble Lipopolymers	177	(1)
Water-Soluble Lipopolymers for IL-12 Gene Delivery to Tumors	178	(1)
Why Secondary Amines for Conjugation of Cholesterol to PEI?	179	(2)
Characterization of T-Shaped Water-Soluble Lipopolymers	181	(4)
Conclusion	185	(2)
Acknowledgments	185	(1)
References	185	(2)
Cyclodextrin-Containing Polymers for Gene Delivery	187	(26)
Suzie Hwang Pun
Mark E. Davis
Biodegradable Polymers
Introduction	187	(1)
Linear, Cyclodextrin-Based Polymers	188	(9)
Cyclodextrins	188	(1)
Polymer Synthesis and Nanoparticle Formulation and Application	189	(2)
Structure-Function Studies	191	(4)
pH-Sensitive Polymers	195	(2)
Polycations Containing Pendant Cyclodextrins	197	(5)
Cyclodextrin-Dendrimer Conjugates	198	(1)
Cyclodextrin-PEI Conjugates	199	(3)
Formulations for In Vivo Applications	202	(5)
Particle Modification by Adamantane Conjugates	202	(1)
Particle Stabilization	202	(1)
Surface Charge Tuning	203	(2)
Targeting	205	(2)
In Vivo Delivery	207	(1)
Biocompatibility of Tf-Particles	208	(1)
Systemic Plasmid Delivery to Tumors	208	(1)
Summary	208	(5)
References	209	(4)
Gene Delivery Using Polyimidazoles and Related Polymers	213	(14)
Sharon Wong
David Putnam
Introduction	213	(1)
The ``Proton Sponge'' Hypothesis	214	(1)
Histidine-Containing Peptides and Polymers	215	(3)
Imidazole Acetic Acid-Containing Polymers	218	(1)
Polyhistidine-Containing Polymers	219	(2)
Vinyl Imidazole-Based Polymers or Polymethacrylates	221	(1)
Future Trends	222	(5)
Acknowledgments	223	(1)
References	223	(4)
Degradable Poly(β-amino ester)s for Gene Delivery	227	(16)
David M. Lynn
Daniel G. Anderson
Akin Akinc
Robert Langer
Introduction	227	(1)
Synthesis of Poly(β-amino ester)s	228	(1)
Degradation and Cytotoxicity	229	(2)
Self-Assembly with Plasmid DNA	231	(2)
Parallel Synthesis and Screening of Poly(β-amino ester) Libraries	233	(2)
Structure-Property Relationships and Biophysical Analysis	235	(1)
Synthesis of Larger Polymer Libraries	236	(1)
pH-Responsive Poly(β-amino ester) Microspheres	237	(1)
Summary	238	(5)
References	240	(3)
Cationic Polyesters as Biodegradable Polymeric Gene Delivery Carriers	243	(12)
Yong-Beom Lim
Yan Lee
Jong-Sang Park
Introduction	243	(1)
Poly(4-hydroxy-l-proline ester) (PHP-ester)	244	(2)
Polymer Synthesis	244	(1)
Molecular Weight Distribution Determination and Degradation	244	(1)
DNA Condensation and Gene Delivery	245	(1)
Poly(α-[4-aminobutyl]-l-glycolic acid) (PAGA)	246	(2)
Polymer Synthesis and PAGA Structure	246	(1)
Degradation and DNA Condensation	247	(1)
Cytotoxicity and Transfection Efficiency	247	(1)
Hyperbranched poly(amino ester) (h-PAE)	248	(2)
Structure and Synthesis	249	(1)
Gene Delivery	249	(1)
Network poly(amino ester) (n-PAE)	250	(3)
Structure and Synthesis	250	(1)
Stability of n-PAE-DNA Polyplex	250	(1)
Gene Delivery	250	(3)
Conclusion	253	(2)
References	253	(2)
Poly(amidoamine)s for Gene Delivery	255	(24)
Paolo Ferruti
Jacopo Franchini
Introduction	256	(1)
Synthetic Aspects	256	(8)
Synthesis and Structure of PAAs	256	(1)
Polymerization Kinetics	257	(1)
Functionalization of PAAs	257	(2)
Amphoteric PAAs	259	(1)
Labeling of PAAs	260	(1)
Radioactive Labeling	260	(1)
PAAs with Amino Pendants and Fluorescent Labeling	260	(3)
PAA Macromonomers	263	(1)
Crosslinked PAAs	263	(1)
PAA-Related Polymers	263	(1)
Physico-Chemical Properties of PAAs	264	(4)
Molecular Weight, Solubility, and Solution Properties	264	(1)
Crystallinity	264	(1)
Thermal Stability	264	(1)
Degradation	264	(3)
Acid--Base Properties of PAAs	267	(1)
Biomedical Applications of PAAs	268	(11)
Applications Mostly Related to Insoluble PAA Derivatives	268	(1)
Neutralization of the Anticoagulant Activity of Heparin, Heparin-Absorbing Resins, and Heparinizable Materials	268	(1)
Soft PAA Hydrogels as Scaffolds for Tissue Engineering	269	(1)
PAAs in Polymer Therapeutics	270	(1)
PAAs as Bioactive and Biocompatible Polymers	270	(1)
Toxicity of PAAs	271	(1)
Body Distribution of PAAs	271	(1)
PAAs as Polymer Carriers for Anticancer Drugs	272	(1)
pH-Mediated Membrane Activity, DNA Complexing, and Transfection Ability of PAAs	273	(1)
PAAs as Promoters of Intracellular Trafficking of Proteins	274	(1)
PAA-Grafted Proteins and Future Trends	275	(1)
References	276	(3)
Cationic Polysaccharides for Gene Delivery	279	(22)
Tony Azzam
Abraham J. Domb
Introduction	279	(2)
Cationic Lipids	280	(1)
Cationic Polymers	280	(1)
Chitosan	281	(3)
Structure, Chemistry, Physico-Chemical Properties, and Application	281	(1)
Chitosan Derivatives	282	(1)
Deoxycholic Acid-Modified Chitosan	282	(1)
Quaternized Chitosan	283	(1)
Chitosan Modified with Hydrophilic Polymers	283	(1)
Galactosylated Chitosan	283	(1)
Chitosan Conjugated with Transferrin, KNOB, and Endosomolytic Proteins	284	(1)
DEAE-Dextran Mediated Transfecting Agent	284	(1)
Polysaccharide-Oligoamine Conjugates for Gene Delivery	285	(7)
Synthesis of Polysaccharide-Oligoamine Conjugates	286	(1)
Condensation Studies of Polysaccharide-Oligoamine Conjugates with DNA	287	(2)
In Vitro Transfection of Polysaccharide-Oligoamine Conjugates	289	(2)
In Vivo Transfection with Dextran-Spermine Vector as a Gene Carrier	291	(1)
Intramuscular Injection of pSV-LacZ Gene Complex	291	(1)
Intravenous Injection of pSV-LacZ Gene Complex	292	(1)
Conclusions	292	(9)
References	294	(7)
Gene Delivery Using Chitosan and Chitosan Derivatives	301	(12)
Gerrit Borchard
Maytal Bivas-Benita
Properties of Chitosan and Chitosan Derivatives	301	(1)
Interactions of Chitosan with Cell Membranes	302	(1)
Preparation of Chitosan-Based Gene Delivery Systems	303	(1)
Polyplexes	303	(1)
Nano- and Microparticles	304	(1)
Transfection Studies	304	(1)
DNA Vaccination	305	(8)
References	307	(6)
Part III Non-Condensing Polymeric Systems
Pluronic® Block Copolymers for Nonviral Gene Delivery	313	(16)
Alexander V. Kabanov
Srikanth Sriadibhatla
Valery Yu. Alakhov
Introduction	313	(3)
Structure and Synthesis of Pluronic® Block Copolymers	316	(1)
Self-Assembly of Pluronic® Block Copolymers	317	(1)
Pluronic® Block Copolymers as Functional Excipients	317	(1)
Pluronics® Enhance Gene Transfer with Polyplexes	318	(1)
Use of Pluronics® as Structural Components in Polyplexes	319	(1)
Pluronics® Enhance Expression in Stably Transfected Cells	320	(1)
Effect of Pluronics® on Expression of Naked DNA In Vivo	320	(2)
Toxicological Considerations	322	(1)
Conclusions	323	(6)
Acknowledgments	323	(1)
References	323	(6)
Use of Poly(N-vinyl pyrrolidone) with Noncondensed Plasmid DNA Formulations for Gene Therapy and Vaccines	329	(16)
Michael Nicolaou
Polly Chang
Mark J. Newman
Introduction	329	(1)
Development of pDNA Vaccines	330	(1)
Delivery Optimization for pDNA Vaccines and Immune-System-Based Therapeutics	331	(1)
Development of Poly(N-vinyl pyrrolidone) (PVP) as a pDNA Delivery Agent	332	(2)
Use of PVP in T-lymphocyte Epitope-Based pDNA Vaccines	334	(2)
Safety Profile of PVP Used as a Drug and Gene Therapy Formulation Excipient	336	(1)
Safety Testing of PVP Formulated EP HIV-1090 pDNA Vaccine for HIV Therapy and Prophylaxis	337	(3)
Summary and Conclusions	340	(5)
Acknowledgments	340	(1)
References	340	(5)
Use of HPMA Copolymers in Gene Delivery	345	(12)
David Oupicky
Introduction	345	(1)
Random Copolymers of HPMA with Cationic Comonomers	346	(2)
Block and Graft Copolymers of HPMA with Polycations	348	(1)
Semitelechelic HPMA Copolymers for Surface Modification of Polyplexes	349	(1)
Multivalent HPMA Copolymers for Surface Modification of Polyplexes and Adenovirus	350	(2)
Surface Modification of Polyplexes	350	(2)
Surface Modification of Adenovirus	352	(1)
Conclusions	352	(5)
References	353	(4)
Part IV Polymeric Nanospheres and Microspheres
Polymeric Nanospheres
Biodegradable Nanoparticles as a Gene Expression Vector	357	(10)
Swayam Prabha
Wenxue Ma
Vinod Labhasetwar
Introduction	357	(1)
Nanoparticles versus Microparticles: The Effect of Size	358	(1)
DNA Incorporation and the Effect of Formulation Conditions	358	(1)
Mechanism of Nanoparticle-Mediated Sustained Gene Transfection	359	(2)
Factors Affecting Nanoparticle-Mediated Gene Transfection	361	(3)
Intracellular Uptake and Endo-Lysosomal Escape	361	(1)
DNA Release	362	(2)
Applications of Nanoparticle-Mediated Gene Transfection	364	(1)
Summary	364	(3)
Acknowledgments	365	(1)
References	365	(2)
Nanoparticles Made of Poly(lactic acid) and Poly(ethylene oxide) as Carriers of Plasmid DNA	367	(20)
Noemi Csaba
Celso Perez
Alejandro Sanchez
Maria Jose Alonso
Introduction	367	(1)
Poly(ethylene oxide) and Derivatives: Physicochemical Properties and Biological Behavior	368	(2)
Poly(ethylene oxide) and Derivatives: New Approaches for Improving Drug Delivery from Nanoparticulate Carriers	370	(1)
Design of Nanoparticles Made of Polyesters, PEO, and Derivatives	371	(3)
Poloxamer/Poloxamine-Coated PLGA Nanoparticles	371	(1)
Poloxamer/Poloxamine-Containing PLGA Nanoparticles	371	(1)
Design of Nanoparticles from Copolymers of PLA and PEG	372	(2)
Nanoencapsulation and Delivery of Oligonucleotides and Plasmid DNA from Nanoparticles	374	(6)
Nanoencapsulation of Oligonucleotides and Plasmid DNA into PLA-PEG Nanoparticles	374	(2)
Nanoencapsulation of Plasmid DNA into PLGA Nanoparticles Containing Poloxamers and Poloxamines	376	(1)
Controlled Release of Plasmid DNA from Nanoparticles of PLA-PLGA and PEO and Derivatives	377	(1)
Stability of Plasmid DNA Released from Nanoparticles of PLA-PLGA and PEO and Derivatives	378	(2)
In Vivo Administration of Plasmid-Containing PLA-PEG Nanoparticles	380	(1)
Concluding Remarks	381	(6)
Acknowledgments	381	(1)
References	381	(6)
Poly(alkylcyanoacrylate) Nanoparticles for Nucleic Acid Delivery	387	(12)
Elias Fattal
Patrick Couvreur
Introduction	387	(1)
Antisense Strategy and the Need for Particulate Formulations	388	(3)
Poly(alkylcyanocrylate) Nanoparticles: Their Potential for ODN Delivery	391	(1)
Delivery of ODN by Polyalkycyanoacrylate Nanospheres	391	(3)
Association of ODN to Nanospheres	391	(1)
In Vitro Stability of ODNs Adsorbed onto Nanospheres	392	(1)
Cell Interactions with ODN-Loaded Nanospheres	393	(1)
In Vitro Pharmacological Activity of Oligonucleotide-Loaded Nanospheres	394	(1)
In Vivo Studies with Oligonucleotide Nanospheres	394	(1)
Delivery of Oligonucleotides by Poly(cyanoacrylate) Nanocapsules	394	(2)
Conclusion	396	(3)
References	396	(3)
Layer-by-Layer Nanoengineering with Polyelectrolytes for Delivery of Bioactive Materials	399	(18)
Dinesh Shenoy
Alexei Antipov
Gleb Sukhorukov
Introduction	399	(2)
LbL Engineering Principles in Design of Colloids with Tailored Architecture	401	(6)
Extension of LbL Sequential Adsorption Technique from Flat Surfaces to Colloidal Particles	401	(1)
Core-Shell Compositions for Drug Delivery Applications	402	(2)
Fabrication of Hollow Capsules by the LbL Approach	404	(1)
Scope and Methods for Loading of Bioactive Materials into Preformed Capsules	405	(1)
Utilization of Donnan Equilibrium	405	(1)
Capturing of Particulate Material, Polymeric Catalyst, or Enzyme	405	(1)
Preparing the Double-Walled Capsules	406	(1)
Extension of LbL Concept for Gene Delivery	407	(2)
DNA Loading by Direct Surface Modification of the Compacted DNA Complex	407	(1)
DNA Encapsulation as Shell Component	408	(1)
Scope for Active Loading of DNA into Preformed Polyelectrolyte Hollow Capsules	409	(1)
Conclusions and Outlook	409	(8)
Acknowledgments	410	(1)
References	410	(7)
Ex Vivo and In Vivo Adenovirus-Mediated Gene Delivery into Refractory Cells via Nanoparticle Hydrogel Formulation	417	(12)
Ales Prokop
Gianluca Carlesso
Jeffrey M. Davidson
Polymer-Driven Adenoviral Gene Delivery	417	(2)
Gene Transfer into Dendritic Cells	419	(3)
Gene Transfer into Pancreatic Cells	422	(1)
In Vivo Gene Delivery	422	(7)
References	423	(6)
Protein Nanoparticles for Gene Delivery	429	(22)
Goldie Kaul
Mansoor Amiji
Introduction	430	(2)
Description of Gene Therapy	430	(1)
Viral versus Non-Viral Vectors	430	(1)
Nanoparticle Systems for Gene Delivery	431	(1)
Protein Nanoparticles	431	(1)
Collagen	432	(4)
Collagen as a Biomaterial	432	(1)
Collagen Nanoparticles for Gene Delivery	433	(1)
Preparation of Nanoparticles	433	(1)
Other Collagen-Based Gene Delivery Systems	434	(1)
Collagen Films and Matrices	434	(1)
Collagen Shields for Gene Delivery	434	(1)
Collagen Sponges	435	(1)
Collagen Hydrogels	435	(1)
Collagen Pellets	435	(1)
Gelatin	436	(2)
Gelatin as a Biomaterial	436	(1)
Gelatin Nanoparticles for Gene Delivery	436	(1)
Gelatin-DNA Coacervates	436	(1)
PEG-Modified Gelatin Nanoparticles	437	(1)
Albumin	438	(2)
Albumin as a Biomaterial	438	(1)
Albumin Nanoparticles for Gene Delivery	439	(1)
Preparation of Albumin Nanoparticles	439	(1)
Additional Examples of Protein Nanoparticles	440	(3)
Lipoproteins	440	(1)
Epidermal Growth Factor	441	(1)
Terplex System	442	(1)
Conclusion	443	(8)
References	443	(8)
Polymeric Microspheres
Gene Delivery Using Poly(lactide-co-glycolide) Microspheres	451	(16)
Mary Lynne Hedley
Introduction	451	(1)
Advantages of PLG Microparticles for Gene Delivery	452	(2)
Protection of DNA	452	(1)
Biocompatibility and Biodegradability	452	(1)
Cell Targeting	453	(1)
Generation of DNA/PLG Microparticle Formulations	454	(3)
Encapsulation of Gene Expression Vectors	454	(1)
Double Emulsion Technique	454	(1)
Spray Drying Technique	455	(1)
DNA-Coated PLG Particles	456	(1)
Particle Internalization and Gene Expression	457	(2)
Immune Activation	459	(4)
Immune Responses Elicited By PLG-Plasmid Formulations	460	(1)
Delivery Systems to Elicit Immune Responses with PLG-Plasmid Particle Formulations	460	(1)
Mucosal Delivery	460	(1)
Parenteral Delivery	461	(1)
Adjuvants for Microparticle-Based Immunization	461	(1)
Nonhuman Primates and Clinical Studies	462	(1)
Summary	463	(4)
References	464	(3)
Polyanhydride Microspheres for Gene Delivery	467	(8)
Yong S. Jong
Camilla A. Santos
Edith Mathiowitz
Introduction	467	(1)
Polyanhydrides	467	(1)
Bioadhesion with Poly(fumaric-co-sebacic anhydride)	468	(1)
Phase Inversion Nanoencapsulation	468	(1)
Improving Particle Uptake for Gene Delivery	469	(1)
Properties of Plasmid DNA	470	(1)
In Vitro Transfection with P(FA:SA) PIN	470	(1)
In Vivo Gene Delivery with P(FA:SA) PIN	471	(1)
Summary	471	(4)
References	472	(3)
Microspheres Formulated from Native Hyaluronan for Applications in Gene Therapy	475	(14)
Yang H. Yun
Weiliam Chen
Introduction	475	(1)
Microspheres as Gene Delivery Vectors	475	(1)
Microspheres Prepared from Hyaluronan	476	(1)
Preparation and Characterization of Hyaluronan Microspheres	476	(5)
Cross-linking of HA Microspheres	477	(1)
HA Microspheres and Size Distribution	478	(1)
Release Kinetics of DNA from HA-DNA Microspheres	478	(2)
Gel Electrophoresis Analysis of DNA Released from HA-DNA Microspheres	480	(1)
In Vitro Cellular Transfection	481	(2)
In Vivo Transfection of Rat Hind Limb Muscles	483	(1)
Conclusions	484	(5)
Acknowledgments	485	(1)
References	485	(4)
Part V Specialized Delivery Systems
Genetically Engineered Protein-Based Polymers: Potential in Gene Delivery	489	(20)
Zaki Megeed
Hamidreza Ghandehari
Introduction	489	(1)
Genetically Engineered Protein-Based Polymers	490	(2)
Synthesis and Characterization of Genetically Engineered Polymers	490	(2)
Biomedical Applications of Genetically Engineered Polymers	492	(2)
Biocompatibility and Biodegradation	492	(1)
Applications of Genetically Engineered Polymers	493	(1)
Gene Delivery Applications for Genetically Engineered Polymers	494	(8)
Controlled Gene Delivery from Silk-Elastinlike Hydrogels	495	(4)
Controlled Release of Adenovirus from Silk-Elastinlike Hydrogels	499	(2)
Gene Delivery from Derivatized Recombinant Polymers	501	(1)
Future Potential of Genetically Engineered Polymers in Gene Delivery	502	(1)
Conclusion and Future Direction	503	(6)
Acknowledgments	503	(1)
References	503	(6)
Glycopolymer Tools for Studying Targeted Nonviral Gene Delivery	509	(14)
Kevin G. Rice
Ji-Seon Kim
Dijie Liu
Introduction	509	(1)
Gene Targeting to C-Type Lectins	510	(2)
Glycosylated Polylysine, PEI, and Peptides	512	(4)
Glycosylated Lipoplexes	516	(1)
Conclusions	516	(7)
Acknowledgment	517	(1)
References	517	(6)
Targeted Gene Delivery via the Folate Receptor	523	(14)
Shih-Jiuan Chiu
Robert J. Lee
Introduction	523	(1)
Gene Therapy for Cancer	524	(1)
The Folate Receptor as a Tumor Marker	524	(1)
Targeted Drug Delivery via the FR	524	(2)
Targeted Gene Delivery via the FR	526	(6)
FR-Targeted Adenoviral Vectors	526	(1)
FR-Targeted Polyplexes	527	(1)
Folate-Poly(L-lysine) (PLL)	527	(1)
Folate-Polyethylenimine (PEI)	528	(1)
Folate-pDMAEMA	529	(1)
FR-Targeted Lipoplexes	529	(1)
FR-Targeted Lipopolyplexes	530	(1)
LPDI	530	(1)
LPDII	531	(1)
Conclusions	532	(5)
References	532	(5)
Transferrin Receptor-Targeted Gene Delivery Systems	537	(20)
Ralf Kircheis
Ernst Wagner
Introduction	538	(1)
Intrinsic Properties of Polycations	538	(4)
DNA Condensation and Electrostatic Interaction with Cell Membrane	538	(1)
Endosomal Release	539	(1)
Polylysine	539	(1)
Polyethylenimine	540	(1)
Migration through Cytoplasm, Nuclear Entry, and Complex Disassembly	541	(1)
Receptor-Mediated Transfection	542	(2)
Transferrin-PEI-DNA Complexes	543	(1)
Specific Ligand-Receptor-Mediated Uptake versus Nonspecific Electrostatic Uptake	543	(1)
Application In Vivo	544	(6)
Systemic Application	546	(1)
Systemically Delivered Polylysine-DNA Complexes	546	(1)
Systemically Delivered PEI-DNA Complexes	547	(1)
Shielding of Polycation-DNA Complexes	547	(1)
PEG Shielding of Polyplexes	547	(1)
Ligand Density	547	(1)
Poly-N-(2-hydroxypropyl)methacrylamide (pHPMA) Shielding	548	(1)
Poloxamer Shielding	548	(1)
Targeted Gene Expression with Shielded Polyplexes	549	(1)
Future Directions	550	(1)
Conclusions	550	(7)
References	551	(6)
Gene Delivery to the Lungs	557	(16)
Berma M. Kinsey
Charles L. Densmore
Frank M. Orson
Introduction	557	(1)
Gene Delivery to the Lung	557	(1)
Polycations	558	(1)
Intravenous Gene Delivery	558	(5)
Introduction	558	(1)
Intravenous Gene Delivery by Polyethylenimine	559	(1)
Ternary Complexes with Polyethylenimine	560	(1)
Polyethylenimine Conjugated to Other Molecules	560	(2)
Gene Delivery by Macroaggregated Albumin--PEI Particles	562	(1)
Cancer Gene Therapy by Intravenous Gene Delivery	562	(1)
Delivery of Genes to the Airways	563	(5)
Introduction	563	(1)
Fluid Instillation of Polyplexes into the Airway	563	(2)
Fluid Phase Airway Gene Delivery with Polyethylenimine-Polyethylene Glycol	565	(1)
Fluid Phase Airway Gene Delivery by Ternary PEI-TAT Peptide-DNA Complexes	565	(1)
Cancer Gene Therapy by Fluid Phase Airway Gene Delivery	565	(1)
Aerosol Gene Delivery to the Lungs by Polyethylenimine	565	(2)
Cancer Gene Therapy by Aerosol Gene Delivery	567	(1)
Genetic Immunization by Aerosol Gene Delivery	567	(1)
Concluding Remarks	568	(5)
References	568	(5)
Cutaneous Gene Delivery	573	(16)
James C. Birchall
Introduction	573	(1)
Disease Targets for Cutaneous Gene Therapy	573	(3)
The Skin as a Barrier to Gene Delivery	576	(1)
Physical Methods for Enhancing Gene Delivery to Skin	577	(4)
Formulation Strategies for Enhancing Gene Delivery to Skin	581	(1)
Polymer-Mediated Gene Delivery to Skin	582	(1)
Targeting Gene Expression in the Skin	583	(1)
Summary	584	(5)
Acknowledgments	584	(1)
References	584	(5)
Enhancement of Wound Repair by Sustained Gene Transfer via Hyaluronan Matrices	589	(22)
Angela P. Kim
Daniel M. Checkla
Don Wen
Philip Dehazya
Weiliam Chen
Introduction	589	(2)
Materials and Methods	591	(3)
Plasmid DNA Gene Constructs	591	(1)
DNA--HA Matrix Formulations	591	(1)
Matrix Morphology Assessment	591	(1)
Assessment of DNA Release Kinetics	592	(1)
Gel Electrophoresis Analyses	592	(1)
Transfections Using Release Samples with DNA Encoding β-Galactosidase	592	(1)
Transfections Using Release Samples with DNA Encoding PDGF-ββ-FLAG	593	(1)
In Vivo Wound Healing Studies	593	(1)
Results and Discussion	594	(13)
Matrix Morphology	594	(1)
Assessment of DNA Release Kinetics	595	(2)
Gel Electrophoresis Analyses	597	(3)
β-Galactosidase Release Sample Transfections	600	(3)
PDGF-ββ-FLAG Release Sample Transfections	603	(4)
In Vivo Wound Healing	607	(1)
Conclusions	607	(4)
Acknowledgments	609	(1)
References	609	(2)
Gene Delivery from Tissue Engineering Matrices	611	(14)
Zain Bengali
Christopher B. Rives
Lonnie D. Shea
Introduction	611	(1)
Matrix-Based Release	612	(4)
Nonviral Delivery	613	(1)
Hydrogels	613	(1)
Synthetic Polymers	614	(1)
Viral Delivery	615	(1)
Solid Phase Delivery	616	(2)
Viral Vector Delivery	617	(1)
Nonviral DNA Delivery	618	(1)
Future Directions and Challenges	618	(7)
References	619	(6)
Gene Therapy Stents for In-Stent Restenosis	625	(34)
Ilia Fishbein
Itay Perlstein
Robert Levy
Delivering Gene Therapy with a Stent	626	(1)
In-Stent Restenosis: The Scope of the Problem	626	(3)
ISR Mechanisms: Platelet Aggregation, Thrombosis, and Thrombus Organization	628	(1)
ISR Mechanisms: SMC Proliferation, Migration, and Matrix Synthesis	628	(1)
Treatment of In-Stent Restenosis	628	(1)
Stent Composition and Design	629	(1)
Coated Stents	629	(1)
Drug-Eluting Stents	630	(1)
Molecular Therapeutics for In-Stent Restenosis: Critical Issues	630	(3)
Plasmid-Mediated Gene Transfer	631	(1)
Replication-Defective Adenovirus-Mediated Gene Transfer	631	(1)
Adeno-Associated-Virus-Mediated Gene Transfer	632	(1)
Retrovirus- and Lentivirus-Mediated Gene Transfer	632	(1)
Antisense-Based Strategies	632	(1)
Cell-Mediated Gene Therapy	633	(1)
Gene Therapy Molecular Targets in Restenosis	633	(4)
Antiproliferative Gene Therapy	633	(3)
Antimigration Gene Therapy	636	(1)
Antithrombotic Gene Therapy	636	(1)
Gene Therapy for Re-Endothelization	636	(1)
Nitric-Oxide-Related Genes: Pleiotropic Therapeutic Effects	637	(1)
Gene Delivery Stents	637	(22)
Plasmid Delivery from a Stent: Bulk Immobilization	638	(1)
Denatured Collagen-Coated Stents Demonstrate Facilitated Plasmid Transfection	639	(3)
Other DNA Delivery Stents	642	(1)
Adenovirus-Mediated Gene Transfer from Stents: The Tethering Strategy	643	(1)
Stent-Tethered Ad Using Antiknob Antibodies	644	(1)
Transduction Using Ad-Delivery Stents in the Pig Coronary Model	644	(15)
Gene Delivery Using BioMEMS	659	(12)
Krishnendu Roy
Introduction	659	(1)
Drug and Gene Delivery Systems	660	(1)
Barriers in Gene Delivery	661	(1)
BioMEMS and Drug Delivery: Current Research	661	(6)
Microfabricated Microneedles	661	(2)
Integrated Delivery Systems	663	(2)
Microchip-Based Delivery Systems	665	(1)
Delivery from Immunoisolated Genetically Modified Cells	666	(1)
Microfabricated Particles for Oral Drug Delivery	667	(1)
Biocompatibility of MEMS Materials	667	(1)
Potential of MEMS in Gene Delivery Applications	668	(3)
References	668	(3)
Index	671