Slide 1: Investigation of Early Outcomes Following AAV Gene Therapy in a Canine Hemophilia Model
Investigation of Early Outcomes Following Adeno-Associated Viral Gene Therapy in a Canine Hemophilia Model (OC 75.2) P. Batty, B. Yates, D. Hurlbut, S. Siso-Llonch, A. Menard, H. Akeefe, T. Christianson, P. Colosi, L. Harpell, S. Liu, A. Mo, A. Pender, G. Veres, A. Winterborn, C. Vitelli, S. Bunting
Slide 2: Disclosures for Paul Batty
Disclosures for Paul Batty In compliance with COI policy, ISTH requires the following disclosures to the session audience: Research Support/P.I. PB has received research funding from BioMarin, Grifols and Octapharma Employee No relevant conflicts of interest to declare Consultant No relevant conflic
Slide 3: Canine Hemophilia A
Canine Hemophilia A • Similar Genotype • Intron-22 inversion like F8 mutation • Similar Clinical Phenotype • Spontaneous bleeding events • Treated on-demand with recombinant canine FVIII • Inhibitor incidence c. 25% • Long-term follow-up >10 years
Slide 4: Study Aims & Outline
Study Aims & Outline AAV5-cFVIII Baseline 3 months 12 months 18 months Liver Biopsy Liver Biopsy Liver Biopsy Liver Biopsy MRI MRI Liver Biopsy: ALT rise ≥2 time upper-limit of normal on repeat testing • Evaluate safety & efficacy of AAV5-cFVIII in a hemophilia dog model • Evaluate hepatic outcomes
Slide 5: Cohort 1: Non-Codon Optimized AAV5-cFVIII
Cohort 1 : Non-Codon Optimized AAV5-cFVIII B-domain deleted canine F8 ITR HLP A1 A2 A3 C1 C2 PA ITR ITR = Inverted terminal repeat. HLP = Hybrid liver promoter PA = Polyadenylation sequence ID Construct Dose Age at Treatment Follow-Up Response (vg/kg) (years) (weeks) AUS2010 HLP-nco-cFVIII-SQ 8.0e13
Slide 6: No Significant FVIII Expression with AAV5-nco-cFVIII
No Significant FVIII Expression with AAV5-nco-cFVIII One Stage FVIII:C WBCT Chromogenic FVIII:C
Slide 7: No Liver Toxicity 12-18 Months Post AAV5-nco-cFVIII
No Liver Toxicity 12 - 18 Months Post AAV5-nco-cFVIII BAN2011 : 12 months post AAV5-nco-cFVIII (2e14 vg/kg) : Age 9.3 years T2 T1+Gad Liver histology – Rare small (3mm) foci of hepatic nodular hyperplasia – likely age related. No other systemic toxicities following AAV5-nco-cFVIII Otherwise unremark
Slide 8: Cohort 2: Codon-Optimized AAV5-cFVIII
Cohort 2 : Codon-Optimized AAV5-cFVIII B-domain deleted canine F8 ITR HLP A1 A2 A3 C1 C2 PA ITR ITR = Inverted terminal repeat. HLP = Hybrid liver promoter PA = Polyadenylation sequence ID Construct Dose (vg/kg) Age at Treatment Follow-Up Current OSA (years) (Weeks) FVIII:C KIN2015 HLP-co-V3-cFVIII
Slide 9: AAV5-co-cFVIII Results in cFVIII Expression and Normal WBCT
AAV5-co-cFVIII Results in cFVIII Expression and Normal WBCT One Stage FVIII:C WBCT Chromogenic FVIII:C
Slide 10: Fall in cFVIII Expression with Elevation in ALT
Fall in cFVIII Expression with Elevation in ALT KIN2015: AAV5-HLP-co-V3-cFVIII : 6.8e13 vg/kg Biopsy Biopsy • Mild post-biopsy ALT elevation at Day 84 • Second ALT elevation at Day 116 : Biopsy performed at Day 119
Slide 11: No Hepatocyte Death or Inflammatory Infiltrate
No Hepatocyte Death or Inflammatory Infiltrate KIN2015: AAV5-HLP-co-V3-cFVIII : 6.8e13 vg/kg Baseline Day 79 Day 119 Pre-dose Day 79 Day 119 Day 79 Day 119 LAL Liver (+)
Slide 12: No Hepatocyte Death or Inflammatory Infiltrate
No Hepatocyte Death or Inflammatory Infiltrate KIN2015: AAV5-HLP-co-V3-cFVIII : 6.8e13 vg/kg Baseline Day 79 Day 119 Pre-dose Day 79 Day 119 LAL Liver (+)
Slide 13: No Increase in GRP78 Expression Following AAV5-cFVIII
No Increase in GRP78 Expression Following AAV5-cFVIII KIN2015: AAV5-HLP-co-V3-cFVIII : 6.8e13 vg/kg GRP78 DAPI Pre-Dose Day 79 Day 119 Vehicle (-) Thapsigargin (+) GRP78 DAPI
Slide 14: No Increase in GRP78 in Hepatocytes Expressing cFVIII
No Increase in GRP78 in Hepatocytes Expressing cFVIII Pre-dose Day 79 Day 119 KIN2015 WT Canine Liver AAV5-HLP-co-V3-cFVIII 6.8e13 vg/kg GRP78 cFVIII DAPI Murine Liver (AAV5-cFVIII) cFVIII cFVIII DAPI merge
Slide 15: Fall in FVIII:C Correlates with Decreased cFVIII mRNA
Fall in FVIII:C Correlates with Decreased cFVIII mRNA Pre-Dose Day 79 Day 119 KIN2015: AAV5-HLP-co-V3-cFVIII : 6.8e13 vg/kg cFVIII DNA cFVIII RNA Tissue Quality Controls UBC dapB
Slide 16: Summary
Summary • Codon Optimized AAV5-cFVIII resulted in therapeutic cFVIII:C expression • No evidence of liver disease or malignancy after 12 – 18 months • First animal model description of elevated ALT & transient loss of cFVIII:C following AAV-FVIII • No hepatocyte cell death, inflammation or upregulati
Slide 17: Acknowledgements
Acknowledgements • Haemophilia colony dogs • Queen’s University • Alex Menard • Lori Harpell • Abbey Pender • David Hurlbut • BioMarin • Sylvia Fong • Bridget Yates Contact: pab7@queensu.ca @paul_batty1