Electroporation in Veterinary Oncology Practice: Electrochemotherapy and Gene Electrotransfer for Immunotherapy

Foreword
Acknowledgements
Contents
Part I: Technology and Science of Electroporation: Fundamentals
	Electroporation: Technology and Science
		1 Introduction
		2 Science: Cells and Tissue Electroporation
			2.1 Electroporation at the Single-Cell Level
			2.2 Electrodelivery in Tissues
		3 Technology
			3.1 Pulses Generators/Pulse Parameters
			3.2 Standard Operating Procedures
		4 New Development/Perspectives
			4.1 Immunotherapy Strategy to Stimulate Immune System
		5 Conclusions
		References
	Electrodes and Electric Field Distribution in Clinical Practice
		1 Introduction
		2 Gross Biological Effects Observed in Electroporation
			2.1 Reversible Electroporation
			2.2 Irreversible Electroporation
			2.3 Thermal Effects
			2.4 Blood Flow Modifying Effects of Pulsed Electric Fields
			2.5 Nerve and Muscle Stimulation
			2.6 Blood-Brain Barrier Disruption
			2.7 pH Changes
		3 Tissue Properties and Computation of Electric Field
			3.1 Physiological State of Tissue
			3.2 Tissue Anisotropy
			3.3 Tissue Perfusion
			3.4 Nonhomogenous Tissue
			3.5 Dynamic Tissue Conductivity
			3.6 Numerical Computation of Electric Field Distribution in Tissue
		4 Electrode Designs and Configurations
			4.1 Noninvasive Electrodes
			4.2 Needle Electrode Arrays (Fixed Geometry)
			4.3 Single Needle Electrodes (Variable Geometry)
			4.4 Finger/Cavity Electrodes
		5 Pulse Parameters (Treatment Protocols)
			5.1 Electrochemotherapy
			5.2 Irreversible Electroporation Ablation
			5.3 Gene Electrotransfer
			5.4 Additional Considerations Regarding Pulse Protocols
		6 Numerical Prediction of Treatment Outcome (Treatment Planning)
			6.1 Cumulative Coverage of Target Tissue
			6.2 The Basics of Patient-Specific Treatment Planning
		7 Conclusion
		References
Part II: Therapeutic Applications with Electroporation in Veterinary Oncology Practice
	Electrochemotherapy in Veterinary Oncology
		1 History of Electrochemotherapy
		2 Electrochemotherapy in Veterinary Oncology
			2.1 Electrochemotherapy for the Treatment of Cutaneous and Subcutaneous Tumors Dogs
				2.1.1 Epithelial Tumors
					Perianal Tumors
					Mammary Adenocarcinoma
					Cutaneous Squamous Cell Carcinoma
					Other Epithelial Tumors
				2.1.2 Mesenchymal Tumors
				2.1.3 Round Cell Tumors
					Mast Cell Tumors
					Other Round Cell Tumors
			2.2 Electrochemotherapy for the Treatment of Oral Tumors in Dogs
				2.2.1 Oral Malignant Melanoma
				2.2.2 Oral Squamous Cell Carcinoma
				2.2.3 Other Oral Tumors
			2.3 Electrochemotherapy for the Treatment of Miscellaneous Tumors in Dogs
			2.4 Electrochemotherapy for the Treatment of Cutaneous and Subcutaneous Tumors in Cats
				2.4.1 Epithelial Tumors
					Squamous Cell Carcinomas
					Mammary Adenocarcinoma
					Other Epithelial Tumors
				2.4.2 Mesenchymal Tumors
					Soft Tissue Sarcomas
				2.4.3 Round Cell Tumors
			2.5 Electrochemotherapy for the Treatment of Oral Tumors in Cats
			2.6 Electrochemotherapy for the Treatment of Miscellaneous Tumors in Cats
			2.7 Procedure
				2.7.1 Patient Selection
					History
					Physical Examination and Diagnostic Work-Up
				2.7.2 Treatment Procedure
					Patient Preparation
					Working Environment and Safety
					Electrode Selection
					Drug Administration
					Electroporation
				2.7.3 Follow-Up Examinations and Retreatments
			2.8 Electrochemotherapy for the Treatment of Sarcoids in Horses
				2.8.1 Procedure
					Patient Selection
					Treatment Procedure
					Patient Preparation
					Working Environment and Safety
					Electrode Selection
					Drug Administration
					Electroporation
					Follow-Up and Retreatments
			2.9 Electrochemotherapy for the Treatment of Tumors in Exotic Pets
		References
	Treating Mast Cell Tumors with Electrochemotherapy
		1 Canine Cutaneous Mast Cell Tumors
			1.1 General Information
			1.2 Clinical Presentation
			1.3 Prognostic Factors
			1.4 Staging and Diagnostic Workup
			1.5 Treatment
				1.5.1 Neoadjuvant Electrochemotherapy (Sole, Pre-surgery)
				1.5.2 Adjuvant Electrochemotherapy (Post-and Intraoperative)
				1.5.3 Gene-Electrotransfer Therapy
		2 Feline Cutaneous Mast Cell Tumor
		Bibliography
	Electrochemotherapy for the Treatment of Transitional Cell Carcinoma in Dogs
		1 Classical Treatment of Tumors of the Bladder and Urethra in Dogs
		2 Electrochemotherapy for the Treatment of Tumors in the Bladder and Urethra
		3 ECT Protocol for the Treatment of Bladder Tumors
		4 ECT Procedure for the Treatment of Bladder Tumors
		5 Follow-Up of Patients Who Underwent ECT for the Treatment of Bladder Tumors
		6 Treatment with COX Inhibitors in Patients Undergoing ECT for the Treatment of Bladder Tumors
		7 ECT Outcomes for Bladder Tumors
		References
	Calcium Electroporation in Veterinary Medicine
		1 Introduction
		2 Calcium Electroporation and How It Is Carried Out
			2.1 Preparation
			2.2 Treatment
				2.2.1 Muscle Contractions
			2.3 Follow-Up
		3 Calcium Electroporation: Background
			3.1 Mechanisms of Action
			3.2 Effects Across Histologies
			3.3 Effects on Normal Versus Malignant Cells
				3.3.1 Mechanisms of Action: Normal and Malignant Cells
			3.4 Calcium Doses and Pulse Application
				3.4.1 Injection Volume, Calcium Concentration, and Calcium Solution
				3.4.2 Generators, Electrodes, and Pulsing Parameters
			3.5 Systemic Effect
		4 Human Clinical Trials
		5 Veterinary Studies with Calcium Electroporation
			5.1 Equine Studies
			5.2 Smaller Domestic Animals: Case Reports
		6 Perspectives
		7 Conclusion
		References
	Irreversible Electroporation Applications
		1 Irreversible Electroporation Theory and Techniques
		2 Treatment Planning
		3 Limitations of IRE and H-FIRE
		4 Clinical Applications of IRE and H-FIRE in Companion Animals with Naturally Occurring Cancers
			4.1 Superficial Soft Tissue Tumors
			4.2 Liver Cancer
			4.3 Brain Cancer
		5 Clinical Applications of IRE and H-FIRE in Preclinical Normal Animal Models or Experimentally Induced Tumors
			5.1 Pancreatic Cancer
			5.2 Prostate Cancer
			5.3 Mammary Cancer
		6 Summary
		References
	Electrochemotherapy as a Multi-Modality Component of Cancer Treatment: Combinations with Surgery, Cryosurgery, Radiation Thera...
		1 Combination with Surgery
			1.1 Excision of the Tumor and Treatment of the Site
			1.2 Flap Reconstruction and ECT
			1.3 Scar Tissue Treatment
		2 Combination with Chemotherapy
		3 Combination with Radiotherapy
		4 Combination with Cryosurgery
		References
Part III: Gene-Electrotransfer and Immunotherapy Applications with Electroporation
	Gene Electrotransfer
		1 Introduction
		2 Mechanism of Gene Electrotransfer
		3 Preclinical Use of GET
			3.1 Preclinical Development for Oncology Applications
			3.2 Preclinical Studies of IL-12 GET in the B16.F10 Mouse Model
		4 Clinical Development for Oncology Applications
			4.1 IL-12 Gene Electrotransfer (GET) Clinical Trials
		5 GET Veterinary Clinical Trials
		6 Summary
		References
	Immunotherapy Applications (Telomerase and HER2) with Gene Electrotransfer
		1 Cancer Vaccines and GET
		2 Genetic Cancer Vaccines and Tumor-Associated Antigens
		3 Preclinical Studies
		4 DNA-GET in Human Clinical Trials
		5 DNA-GET in Veterinary Clinical Trials
		6 Conclusions
		References
	Interleukin-12 Gene Electrotransfer in Veterinary Oncology
		1 Introduction
			1.1 Antigen Presenting Cells
			1.2 Cytotoxic Immune Cells
			1.3 Interleukin-12
		2 IL-12 Gene Electrotransfer
			2.1 IL-12 Gene Electrotransfer Studies on Mice
			2.2 IL-12 Gene Electrotransfer Studies in Companion Dogs with Cancer
				2.2.1 Effectiveness of IL-12 gene Electrotransfer ALONE or in Combination with ECT
				2.2.2 Immune Response after IL-12 Gene Electrotransfer Alone or in Combination with ECT
				2.2.3 Safety of IL-12 Gene Electrotransfer Alone or in Combination with ECT
				2.2.4 Health-Related Quality of Life
				2.2.5 Plasmid Shedding
		3 Future Directions and Conclusions
			3.1 Optimization of Plasmid DNA
			3.2 Course of Treatment
		References
	Canine Melanoma and Osteosarcoma Immunotherapy by Means of In Vivo DNA Electroporation
		1 Canine Malignant Melanoma and Osteosarcoma: An Overview
		2 Immunotherapeutic Approaches in Canine Malignant Melanoma and Osteosarcoma Oncology
			2.1 Nonspecific Immunotherapy
			2.2 Checkpoint Inhibitors
			2.3 Monoclonal Antibodies Against Tumor-Associated Antigens
			2.4 CAR-T Cells
			2.5 Vaccines
		3 DNA Vaccines
		4 Application of Electroporation to DNA Vaccination
		5 Case Study: DNA Vaccination with In Vivo Electroporation in Canine Oral Malignant Melanoma and Osteosarcoma
			5.1 CSPG4
			5.2 CSPG4 in canine OMM and OSA
			5.3 CSPG4 Electrovaccination in Canine OMM and OSA
		6 Conclusions
		References
Part IV: Evolution of the Field and New Applications with Electroporation-Based Treatments: Outlook
	Electrodes for Unique Anatomical Access in Electroporation
		1 Introduction
		2 The Electrodes
		3 Single Needle Electrode
		4 The Curved Electrode
			4.1 Finger Electrode
		5 SN-HFIRE Electrode
		6 Brain Electrode
		7 Endo-Ve
		8 Pulsed-Field Ablation System for Cardiac Ablation
			8.1 Placing independent needles
		References
	New Electrodes and Treatment Planning for Deep-Seated and Intraluminal Localized Tumors
		1 Background
		2 ECT in Preclinical Studies of Deep Tumors
		3 The Expandable and Deployable Electrodes
			3.1 Our Experience on the Pig Model with Expandable and Deployable Electrodes
		4 Bipolar Electrodes
			4.1 Our Experience on the Pig Model with Bipolar Electrodes
		5 Treatment Planning
		6 Conclusions
		References
	Advancing Electroporation Systems
		1 Background
		2 Concepts of Electroporation Systems
		3 Generator Structure
			3.1 Type of Signal Generators (Rebersek and Miklavcic 2010)
			3.2 Design of a Generator: The Genedrive Electroporation System
		4 Non-Tumor Electroporation Applications
			4.1 Cardiac Ablation
			4.2 Electrosclerotherapy of Vascular Malformations
			4.3 Gene Therapy
		5 Conclusions
		References