Slide 1: Systemic Delivery of AAV9.LAMP2B for the Treatment of Danon Disease
Systemic Delivery of AAV9.LAMP2B for the Treatment of Danon Disease: Toxicology Studies in Mice and Cynomolgus Monkeys Annahita Keravala, Ph.D. Associate Vice President, AAV Platform
Slide 2: Disclosure Information
Disclosure Information • Pavan Battiprolu, David Ricks, Simon Moore, Sanchali Kasbekar, Kinnari Patel, Jonathan D. Schwartz, Gaurav Shah, and Annahita Keravala are employees of Rocket Pharmaceuticals, Inc. • Ana-Maria Manso is a consultant for Rocket Pharma • Eric Adler is a shareholder in Rocket Ph
Slide 3: Cautionary Statement Regarding Forward-Looking Statements
Cautionary Statement Regarding Forward-Looking Statements Various statements in this presentation concerning Rocket’s future expectations, plans and prospects, including without limitation, Rocket’s expectations regarding the safety, effectiveness and timing of product candidates that Rocket may dev
Slide 4: Danon Disease (DD)
Danon Disease (DD) • Danon disease is a devastating multi-system disorder primarily affecting cardiac tissue • 95% of patients have severe cardiomyopathy and die from progressive heart failure • Heart transplant is a treatment option, which is not curative & is associated with considerable morbidity
Slide 5: AAV9 Vector Shows Consistent & Strong Cardiac Tropism Across Species
AAV9 Vector Shows Consistent & Strong Cardiac Tropism In Several Studies Across Different Species Disorder & Dose Species Results Sponsor Reference Vector LGMD2A 3E+13 vg/kg NHP 8-80-fold higher transduction in cardiac vs. Genethon Lostal et al AAV9.hCAPN3 skeletal muscle (ASGCT 2018) Non-specific 3
Slide 6: Study To Assess Efficacy of AAV9.LAMP2B (RP-A501) in LAMP2 Knockout Mouse Model
Study To Assess Efficacy of AAV9.LAMP2B (RP-A501) in LAMP2 Knockout Mouse Model of Danon Disease Intravenous injection: AAV9.LAMP2B 6
Slide 7: Dose-dependent Expression of LAMP2 and Decreased LC3-II in Heart
Dose-dependent Expression of LAMP2 and Decreased LC3-II in Heart Western Blot hLAMP2 LC3-II mLAMP2 **** **** **** 4 **** 2.0 (Normalized to GAPDH) **** (Normalized to GAPDH) *** hLAMP2 Intensity LC3-II Intensity 3 **** 1.5 2 ** 1.0 1 0.5 0 0.0 Untx Untx WT PBS 1e13 5e13 1e14 2e14 Untx WT PBS 1e13 5e
Slide 8: Restoration of Cardiac Ultrastructure and Systolic & Diastolic Function
Restoration of Cardiac Ultrastructure and Systolic & Diastolic Function Electron Microscopy Invasive Hemodynamics Cardiac Contractility Cardiac Relaxation Cardiac Contractility Cardiac Relaxation ** 10000 ** **** 0 * **** * dP/dt max (mmHg/s) dP/dt min (mmHg/s) 8000 *** **** ** -2000 6000 ** -4000 4
Slide 9: IND-Enabling Studies to Assess Safety of AAV9.LAMP2B (RP-A501)
IND-Enabling Studies to Assess Safety of AAV9.LAMP2B (RP-A501) Toxicology/Biodistribution studies were conducted: I. A GLP study performed in wild-type C57BL/6 mice • Doses: 3×1013, 1×1014, 3×1014 vg/kg • Assessment timepoints: 30, 91, and 180 days post infusion II. NHP study performed in cynomolgus
Slide 10: Toxicology Assessment of RP-A501 in Mice at Days 30, 91, and 180
Toxicology Assessment of RP-A501 in Mice at Days 30, 91, and 180 Assessment Outcome Clinical blood chemistry No relevant dose-related findings in clinical chemistry panel Hematology No relevant dose-related changes at any time-point Organ weight No adverse organ weight changes related to dosing Hist
Slide 11: Biodistribution of RP-A501 in Mice at Days 30 & 91
Biodistribution of RP-A501 in Mice at Days 30 & 91 LAMP2B mRNA ➢ Differential distribution of vector genomes (vg) was noted ➢ Highest levels of mRNA were measured in heart, which were stable between day 30 and 91 timepoints 11
Slide 12: Toxicology Assessment of RP-A501 in Non-human Primates at Day 102
Toxicology Assessment of RP-A501 in Non-human Primates at Day 102 Assessment Outcome Moribundity/mortality No clinical signs of toxicity manifesting as moribundity or mortality Electrocardiography No changes in heart rhythm, ST segment, waveform morphology, MEA, or interval measures Clinical patholo
Slide 13: Immunogenicity Assessment of RP-A501 in NHPs
Immunogenicity Assessment of RP-A501 in NHPs NAb TAb ADA AAV9 NAb IC50 Titer (Cut Point of 3.156) Titer (Cut point of 1.283) Group Animal ID Group Animal ID Group Animal ID Pre-Test Day 30 Day 90 Pre-dose Day 30 Day 90 Pre-dose Day 30 Day 90 Control 2750 0 443 162 Control 2750 <10 100 <10 Control 27
Slide 14: VCN and mRNA Assessment for RP-A501 Biodistribution in NHPs
VCN in NHPs at Day 102 Br a in WPRE mRNA Ce mRNA copies/ total RNA mg of copies per mg total RNA B r reb mRNA 1010 105 106 107 108 109 ai el n lu Br Fro m B Br ra ain ntal ai in Hi n p ➢ High levels of transduction (mRNA) were observed in the heart Oc Me p. Br . c du C ll Br ain or a ai Pa tex n Te
Slide 15: RP-A501-Mediated hLAMP2 Protein Expression in NHPs at Day 2
RP-A501-Mediated hLAMP2 Protein Expression in NHPs at Day 102 Western Blot LAMP2 GAPDH Right Left Vehicle 0.3 RP-A501 ** ng LAMP2 per mg of Total Protein *p<0.05, **p<0.01 (normalized based on BCA) LAMP2 assessment ** ** based on total protein+ 0.2 loaded on gel * 0.1 ➢ Higher levels of transgenic h
Slide 16: Key Takeaways: RP-A501 is Effective & Safe in Preclinical Studies
Key Takeaways: RP-A501 is Effective & Safe in Preclinical Studies ✓ AAV9.LAMP2B effectively transduces the heart (key target organ for DD). High VCNs, mRNA, as well as protein were noted in cardiac tissue of dosed mice and NHPs ✓ AAV9.LAMP2B confers direct benefit to cardiac function and improves ul