Peter M Thulé1,2, Adam G Campbell1, Dingwu Jia1, Yulin Lin1, Shou You3, Sara Paveglio4, Darin E Olson1,2, Miroslaw Kozlowski5. 1. Section Endocrinology and Metabolism, Atlanta VA Medical Center, Decatur, GA, USA. 2. Division of Endocrinology, Metabolism, & Lipids, Emory University School of Medicine, Emory University, Decatur, GA, USA. 3. Department of Endocrinology, Second Xiangya Hospital, Central South University, Changsha, China. 4. UConn Health Center, Farmington, CT, USA. 5. Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Abstract
BACKGROUND: Insulin self-administration is burdensome and can produce dangerous hypoglycemia. Insulin gene therapy may improve and simplify the treatment of diabetes mellitus. In rats, metabolically responsive hepatic insulin gene therapy (HIGT) delivered by adenovirus normalizes random blood sugars but with a limited duration. To prolong glycemic control, we delivered a metabolically regulated insulin transgene by adeno-associated virus (AAV). METHODS: We administered increasing doses of self-complementary (SC), pseudotyped AAV8 expressing the (GlRE)3 BP1-2xfur insulin transgene to streptozotocin-diabetic CD-1 mice, and monitored blood sugar and body weight. We also compared responses to intraperitoneal glucose and chow withdrawal, assessed for viral genomes in liver by Southern blotting, and measured hepatic glycogen. RESULTS: Glucose lowering required the combination of SC genomes and AAV capsid pseudotyping. HIGT controlled glycemia in diabetic mice (DM) for > 1 year. However, glycemic responses were variable. Approximately 30% of mice succumbed to hypoglycemia, and approximately 30% of mice again became hyperglycemic. During an intraperitoneal glucose tolerance test, blood sugars declined to normal within 180 min in HIGT-treated DM compared to 90 min in control mice. Hypoglycemia was common among HIGT-treated mice during a 24-h fast. However, HIGT mice lost less weight than either diabetic or nondiabetic controls as a result of increased water intake. HIGT treatment reduced the hepatic glycogen content of fed mice. CONCLUSIONS: Our studies demonstrate the possibility for long-term glycemic correction following AAV-mediated HIGT in mice. However, the dose-response relationship is irregular, and metabolic responsiveness may be less than that observed in rats.
BACKGROUND: Insulin self-administration is burdensome and can produce dangerous hypoglycemia. Insulin gene therapy may improve and simplify the treatment of diabetes mellitus. In rats, metabolically responsive hepatic insulin gene therapy (HIGT) delivered by adenovirus normalizes random blood sugars but with a limited duration. To prolong glycemic control, we delivered a metabolically regulated insulin transgene by adeno-associated virus (AAV). METHODS: We administered increasing doses of self-complementary (SC), pseudotyped AAV8 expressing the (GlRE)3 BP1-2xfur insulin transgene to streptozotocin-diabetic CD-1 mice, and monitored blood sugar and body weight. We also compared responses to intraperitoneal glucose and chow withdrawal, assessed for viral genomes in liver by Southern blotting, and measured hepatic glycogen. RESULTS:Glucose lowering required the combination of SC genomes and AAV capsid pseudotyping. HIGT controlled glycemia in diabeticmice (DM) for > 1 year. However, glycemic responses were variable. Approximately 30% of mice succumbed to hypoglycemia, and approximately 30% of mice again became hyperglycemic. During an intraperitoneal glucose tolerance test, blood sugars declined to normal within 180 min in HIGT-treated DM compared to 90 min in control mice. Hypoglycemia was common among HIGT-treated mice during a 24-h fast. However, HIGT mice lost less weight than either diabetic or nondiabetic controls as a result of increased water intake. HIGT treatment reduced the hepatic glycogen content of fed mice. CONCLUSIONS: Our studies demonstrate the possibility for long-term glycemic correction following AAV-mediated HIGT in mice. However, the dose-response relationship is irregular, and metabolic responsiveness may be less than that observed in rats.
Authors: Petra Baum; Severin Koj; Nora Klöting; Matthias Blüher; Joseph Classen; Sabine Paeschke; Martin Gericke; Klaus V Toyka; Marcin Nowicki; Joanna Kosacka Journal: Int J Mol Sci Date: 2021-02-04 Impact factor: 5.923
Authors: Asha Recino; Shu Uin Gan; Kian Chuan Sia; Yvonne Sawyer; Jenny Trendell; Richard Kay; Fiona M Gribble; Frank Reimann; Rob Foale; Maria Notaridou; Nick Holmes; Andrew Lever; Kok Onn Lee; Amit Nathwani; Anne Cooke; Roy Calne; Maja Wallberg Journal: Gene Ther Date: 2018-12-04 Impact factor: 5.250