Adenoviral Vectors for Gene Therapy - D. Curiel, J. Douglas (AP, 2002) WW

673 Pages·2002·43.53 MB·English

ADENOVIRAL VECTORS
FOR GENE THERAPY

ese

a aad “ “
DAVID T. CURIEL | JOANNE T. DOUGLAS
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Contents

Contributors rail

Preface

CHA

TERI

Adenovirus Structure
Phoebe L. Stewart

|. Introduction 1
|. Molecular Composition 2
|. Structure of the Intact Virion 3

Structure of the Capsid Components
A. Hexon, Polypeptide I 4
B. Penton Base, Polypeptide IIL
C. Fiber, Polypeptide IV 8
D. Polypeptide Ila 10

E. Polypeptide VI 10

F. Polypeptide VIII n

G. Polypeptide IX n

Material
V. Core Structure 12

VI. Adenovirus Protease 2
VIL. Summary 13
References “4

CHAPTER 2

Biology of Adenovirus Cell Entry
Glen R. Nemerow
|. Pathway of Adenovirus Cell Entry 9
|. Cell Receptors Involved in Attachment 2
A, CAR 2
B. Other Adenovirus Receptors 23
Ill. Adenovirus Internalization Receptors 24
A. Role of av Integrins as Coreceptors 24
B. Structural Features of Penton Base-av Integrin
Association 26
C. Signaling Events Associated with Adenovirus
Internalization 26
IV. Virus-Mediated Endosome Disruption and Uncoating 29
‘A. Role of Penton Base and av Integrins 29
B. Role of the Adenovirus Cysteine Protease 30

V. Beyond the Endasome: Tracking of Viel Copsics nd Inport of
Viral DNA into the Nucleus

A. Intracytoplasmic Transport ny Viral Capsids 30
B. Docking at the Nuclear Pore and Translocation of Viral
DNA 31
VI. Conclusions 31
References 32

CHA

Adenovirus Replication

Jared D. Evans and Patrick Hearing
|. Introduction 39
. Classification 40
Ill, Genome Organization 40
IV. Virus Infection a2
V. Early Gene Expression 42
Vi. Early Region 1A (E1A) 43
Vil. Early Region 18 (E1B) 46
Vill. Early Region 2 (E2) a7
IX. Early Region 3 (E3) 49
X. Early Region 4 (E4) 52
XI, Viral DNA Replication 55
Xil. VA RNA Genes 59
XIll. Late Gene Expression and Virus Assembly 39
XIV. Vector Design 62
XV. Conclusion 63
References 64

CHAPTER 4

Adenoviral Vector Construction I: Mammalian Systems

Philip Ng and Frank L. Graham
|. Introduction n
A. Adenovirus Biology n
B. Adenovirus Vectors 73
C. Early Methods of Constructing Recombinant
Adenoviruses 73

Il. The Two-Plasmid Rescue System 75
A. Development of the Two-Plasmid Rescue System
B. Refinements to the Two-Plasmid Rescue Method
C, The Ad Genomic Plasmid 82
D. The Shuttle Plasmid 82

Il. Protocols for the Two-Plasmid Rescue System 85
A. Preparation of Plasmid DNA 85
B. Cell Culture 388
C. Cotransfection 89

7
79

D. Analysis of Recombinant Vectors and Preparation of Working

Vector Stocks 93
Contents

E, Titration of Adenovirus 94
Preparation of High-Titer Viral Stocks (Crude
Lysate) 95

G. Purification of Adenovirus by CsCl Banding 97

H. Characterization of Adenoviral Vector
Preparations. = 99.

I. Alternative Procedures to Expedite Vector
Production 101
References 101

CHAPTERS

Adenoviral Vector Construction II: Bacterial Systems
M. Lusky, E. Degryse, M. Mehtali, and €. Chartier
|. Introduction 105
I. Generation of Ad: Traditional Approaches 106
Ill. Generation of Ad: Bacterial Systems 107

IV. Homologous Recombination in E. coli 108

V. Homologous Recombination with Linear Ad Vector Genome
Plasmids 109

VI. Homologous Recombination with Circular Ad Vector Genome
Plasmids 6

VII. Ad Vector Construction by Transposon-Mediated
Recombination ne

Vill. Ad Vector Construction by in Vitro Ligation ng
IX. Conclusion 121
References 123

CHAPTER 6

Propagation of Adenoviral Vectors: Use of PER.C6 Cells
W. W. Nichols, R. Lardenoije, B. J. Ledwith, K. Brouwer, S. Manam, R. Vogels,
D. Kaslow, D. Zuidgeest, A. J. Bett, L. Chen, M. van der Kaaden,
5. M. Galloway, R. B. Hill, S. V. Machotka, C. A. Anderson, J. Lewis,
D. Martinez, J. Lebron, C. Russo, D. Valerio, and A. Bout
|. Introduction 129
Contents

M

mM.

A. Scope of the Chapter 129
B. Adenoviruses 130
C. Adenovirus Replication 131

Cells Expressing E1 of Adenovirus. 134
A. Transformation of Cells by El of

Adenovirus 134
B. El-Expressing Cell Lines for Adenoviral Vector
Production 135

PER.C6 Prevents RCA during Vector Production 136

A. RCA 136

B. PER.C6: Absence of Sequence Overlap Eliminates RCA
Generation 137

Occurrence 139

C. Frequency of R

IV. Production of Adenoviral Vectors 141

A. Vector Stability 141
B. The Production Process 143

C. Yields of Adenoviral Vectors 145

D. Scale of Adenoviral Vector Production 145

V. Safely Considerations of PER.C6 146

A. QC Testing of PER.C6 Cells for Use in the Manufacture of
Biologicals and Vaccines 146

B, Tumorigenicity 150

C. Prion-Related Issues 156

D. Genetic Characterization of PER.C6 Cells 157

VI. Conclusions 159

CHA

References 160

PTERZ

Purification of Adenovirus
Paul Shabram, Gary Vellekamp and Carl Scandella

|. Introduction 167
A. The Physical Characteristics of the Adenovirus Particle in
Solution 167
B, Features of the Milieu 170
. Summary of Characteristics 173
Il. Recovery and Purification of Adenoviral Particles 173
A. Harvest Methods W733
B. Lysis Methods 175
C. Clarification 179
D. Purification 181
E. Buffer Exchange 189

Il. Analytical Methods for Process Development and Process
Tracking 190

A. Plaque-Forming Titer Assays 190

B, Adenovirus 96-Well Titer Plate Assay 191
C. Flow Cytometry 194
D.

|. Particle Concentration Determination by Ultraviolet
Absorbance 194

E, Analytical Reverse-Phase HPLC 195
F. Analytical Anion-Exchange HPLC 196
Formulation and Stability 196
Conclusions 200
References 200

<2

CHAPTER 8

Targeted Adenoviral Vectors I: Transductional Targeting
Victor Krasnykh and Joanne T. Douglas
|. Introduction 205
Il. The Pathway of Adenoviral Infection 206
Ill. Strategies and Considerations 207
IV. Conjugate-Based Targeting 209

A. Bispecific Chemical Conjugates 209

B. Bispecific Recombinant Fusion Proteins 213
C. Bispecific Peptides 215

D. Polymer-Mediated Coupling of Ligands to Ad Capsid

Proteins 215
E. Biotinylated Ad/Avidin Bridge/Biotinylated
Ligand 216
V. Genetic Targeting 216
A. Ad-Targeting Strategies Involving Genetic Manipulations of

the Fiber Protein 217
B. Ad-Targeting Strategies Involving Genetic Manipulations of
the Hexon and Penton Base Proteins 231

VI. Transductionally Targeted Ad Vectors for Clinical Gene Therapy
Applications 232

VII. Conclusion 235
References 236

CHAPTER 9

Targeted Adenoviral Vectors II: Transcriptional Targeting
Sudhanshu P. Raikwar, Chinghai H. Koo, and Thomas A. Gardner
|. Introduction: Rationale of Transcriptional Targeting 207
II. Regulation of Transcription in Eukaryotes 248
Molecular Organization of DNA 248
‘The Central Dogma 248
‘Transcription 249
Mechanism of Transcription 251
. Structural Motifs 253

Regulation of Adenoviral DNA Transcription
Process 256

Il. Approaches of Transcriptional Regulation 256
A. Prior Rationale Universal Promoters 256
B. Current Rationale of Tissue-Specific Promoters 257
C. MNICA9 Promoter 268
D. Inducible Transcription 269
IV. Enhanced Control of Transgene Expression 271
A. Safety Improvements an
B. Potency Concerns 272
V. Future Directions 272
A. Enhancement of Weak But Specific Promoters. 272

mmo Oe >
B. Improving Specificity with Multiple Promoter

Segments 274
C. Tumor-Specific Oncolysis 274
D. Combined Targeting Approaches 274
Vi. Summary 274
References 275

CHAPTER 10

Development of Attenuated Replication Competent
Adenoviruses (ARCAs) for the Treatment of Prostate Cancer
Daniel R. Henderson and De-Chao Yu

Introduction 287
ARCAs for Prostate Cancer: CV706 and
CVv787 290

A. Adenovirus: Gene Expression and
Regulation 290

B. Tissue Specificity of ARCA 291

C. Antitumoral Efficacy of ARCA 295

D. Mechanism for Cell-Killing of ARCA 296

. Synergy of ARCA and Conventional Therapy 298

A. Synergy of CV706 and Irradiation 299

B. Synergy of CV787 and Chemotherapy 302
Toxicity of Intravenously Administered ARCAs in the Absence or
Presence of Docetaxel 305

Effects of Preexisting Adenovirus Antibody on Latina Activity and
Immunoapheresis for Human Therapy

A. Preexisting Adenovirus Antibodies rane it yeeale Toxicity

and Antitumor Activity 309

B. SIAPA: Screening and Immunoapheresis of Preexistent
Antibody for Monitoring and Removing Preexistent AdS
Antibodies from Blood 313

Clinical Development of CV706 and CV787 314

A. CV706 Phase I/II Trial for Locally Recurrent Prostate
Cancer 314

B. Factors Impacting Clinical Efficacy and Safety 317