Slide 1: AAV9.LAMP-2B Reverses Metabolic and Physiologic Multiorgan Dysfunction in a Murine Model of Danon Disease
AAV9. LAMP-2B Reverses Metabolic and Physiologic Multiorgan Dysfunction in a Murine Model of Danon Disease Ana Maria Manso, Emily Gault, Sherin Hashem, Bradley Nelson, Elizza Villarruel, Pavan Battiprolu, Annahita Keravala, Yusu Gu, Nancy Dalton, Kirk Hammond, Kirk Peterson, Eric Adler UCSD School o
Slide 2: Disclosure Information
Disclosure Information • Ana Maria Manso is a consultant for Rocket Pharmaceuticals. • Pavan Battiprolu and Annahita Keravala are employees of Rocket Pharmaceuticals • Eric Adler is a shareholder of Rocket Pharmaceuticals.
Slide 3: Danon Disease: Clinical Manifestations
Danon Disease: Clinical Manifestations • Lethal X-linked disorder caused by mutations in the Lysosomal Associated Membrane Protein 2 (LAMP-2) gene, a lysosomal protein critical for autophagy. • Cardiomyopathy, skeletal myopathy, and intellectual disability. • Other less prevalent symptoms include re
Slide 4: Autophagy
Autophagy Danon Disease LAMP-2 LAMP-2 LAMP-2 LC3 II LC3 II LC3 II LC3 II LC3 II LC3 II LC3 II LC3II LC3 II Impaired autophagosome-lysosome fusion impairs autophagy flux in Danon disease
Slide 5: LAMP-2 isoforms
LAMP-2 isoforms Chi et al., 2018 LAMP-2B is the predominant isoform expressed in cardiomyocytes and has been postulated to be critical in the pathogenesis of Danon disease.
Slide 6: Aim: Evaluate efficacy of AAV9-LAMP-2B gene therapy in a mouse model of Danon disease
Aim: Evaluate the efficacy of gene therapy with adeno- associated viral 9 (AAV9-LAMP-2B) vector in a mouse model of Danon disease.
Slide 7: AAV9-LAMP-2B treatment in LAMP-2 KO mice study
AAV9-LAMP-2B treatment in LAMP-2 KO mice study 12 weeks √ 24 weeks √
Slide 8: Dose-dependent LAMP-2B expression and autophagic flux in heart tissue
Administration of AAV9.LAMP2B showed dose-dependent expression of mRNA and human LAMP-2B protein in heart tissue from LAMP2 KO mice together with an improvement in autophagic flux (LC3 II levels) 24 weeks post-injection LAMP-2 LC3 II LC3 II LC3 II LC3 II LC3 II LC3 II LC3II *P<0.05, **P<0.01, ***P<0
Slide 9: Improvement in accumulation of autophagic structures in heart
AAV9.LAMP-2B administration was associated with an improvement in the accumulation of autophagic structures in hearts of LAMP-2 KO mice.
Slide 10: Cardiac contractility and relaxation improved dose-dependently
Cardiac contractility and relaxation were also improved in a dose-dependent manner in the AAV9.LAMP2B treated LAMP2 KO mice compared to PBS controls 24 weeks post-injection Cardiac Contractility Cardiac Relaxation ** P<0.01, ***P<0.001, ****P<0.001 vs WT # P<0.05, ##P<0.01, ###P<0.001 vs PBS
Slide 11: Dose-dependent LAMP-2B expression and autophagic flux in liver
Administration of AAV9.LAMP-2B showed dose-dependent expression of mRNA and human LAMP-2B protein in liver tissue from LAMP-2 KO mice together with an improvement in autophagic flux (LC3 II levels) # ** P<0.05 vs PBS P<0.01, vs WT
Slide 12: Improvement in autophagic structures in liver
AAV9.LAMP2B administration was associated with an improvement in the accumulation of autophagic structures in livers of LAMP-2 KO mice.
Slide 13: Serum ALP and ALT levels in LAMP-2 KO mice
AAV9.LAMP-2B administration was associated with an improvement in the serum levels of ALP and ALT in LAMP-2 KO mice. Alkaline phosphatase Alanine aminotransferase ** P<0.01, ****P<0.001 vs WT # P<0.05, ##P<0.01, ####P<0.0001 vs PBS
Slide 14: Dose-dependent LAMP-2B expression in skeletal muscle
Administration of AAV9.LAMP-2B showed dose-dependent expression of human LAMP-2B protein in skeletal muscle tissue from LAMP-2 KO mice together with an improvement in autophagic flux (LC3 II levels) * P<0.05 ,**P<0.01 vs WT #P<0.05, ##P<0.01 vs PBS
Slide 15: Improvement in autophagic structures in skeletal muscle
AAV9.LAMP-2B administration was associated with an improvement in the accumulation of autophagic structures in skeletal muscle of LAMP-2 KO mice.
Slide 16: Summary
Summary • Administration of AAV9.LAMP-2B showed dose-dependent expression of human LAMP-2B transcript and protein in heart, liver and skeletal muscle. • AAV9.LAMP-2B administration was associated with an improvement in autophagic flux (LC3 II levels) as well as in the accumulation of autophagic stru
Slide 17: Acknowledgments
Acknowledgments Dr. Eric Adler Emily Gault Elizza Villarruel Yusu Gu Paul Bushway Angel Soto-Hermida Chao Chen Sherin Hashem Bradly Nelson Michela Brambatti Pavan Battiprolu Annahita Keravala Jonathan Schwartz Gaurav Shah H. S. Lopez Family Foundation founded by Czarina and Humberto S. Lopez