uniQure N.V. · NASDAQ:QURE
uniQure — ASGCT 2019
20 slides from uniQure's conference presentation, Aug 13, 2019
Slide 1: Development of an AAV5 gene therapy for Fabry disease
Development of an AAV5 gene therapy for Fabry disease Ying Poi Liu, PhD uniQure Biopharma B.V. Amsterdam, The Netherlands
Slide 2: Forward-looking statements
This presentation contains forward-looking statements. All statements other than statements of historical fact are forward-looking statements, which are often indicated by terms such as “anticipate,” “believe,” “could,” “estimate,” “expect,” “goal,” “intend,” “look forward to,” “may,” “plan,” “poten
Slide 3: Fabry disease: a lysosomal storage disease
Fabry disease: a lysosomal storage disease - X-linked genetic disorder - Deficiency of α-galactosidase A (α-Gal A or GLA) - Females also suffer from Fabry and severity depends on X-inactivation despite having GLA activity in the plasma - Systemic accumulation of substrate; Gb3 and LysoGb3 in plasma,
Slide 4: What is cross correction and why is it important?
What is cross correction and why is it important? Disadvantages of ERT: • Poor cross correction of GLA • Heterozygous females are also symptomatic • Thus, unaffected cells produce GLA but uptake into lysosomes via the Mannose 6- phosphate receptor is poor • In MPS II, there are asymptomatic carriers
Slide 5: uniQure's approach: modified NAGA
uniQure’s approach: modified NAGA Licenced from Prof. Sakuraba, Tokyo Novel Approach • Expression of modified NAGA (modNAGA) using AAV5 vector (constant supply) • ModNAGA has GLA activity and is able to reduce LysoGb3 accumulation More stable in blood and low pH modNAGA is Expression of active in th
Slide 6: Two approaches: liver specific or constitutive promoter
Two approaches: liver specific or constitutive promoter AAV5 ModNAGA L1 Liver produces and secretes protein can ModNAGA be taken up into target organs C1 Constitutive protein NAGA expressoin from target L1 organs or coNAGA SV40pA C1 spNAGA GLA signal peptide DELIVERING GENE THERAPY TO PATIENTS Fabry
Slide 7: Studies to show proof of concept of modNAGA in vitro and in vivo
Studies to show proof of concept of modNAGA in vitro and in vivo In vitro, cells Wt mice Fabry mice GLA activity GLA activity GLA activity M6P-receptor (plasma and target organs) LysoGb3 mediated uptake (plasma and target organs) DELIVERING GENE THERAPY TO PATIENTS Fabry program | 7
Slide 8: Expression of modNAGA results in GLA activity in cells and cellular uptake via M6P-receptor
Expression of modNAGA results in GLA activity in cells and cellular uptake is mediated via M6P-receptor Plasmid transfection modNAGA GLA activity in GLA activity in +/- M6P expression cells supernatant no M6P no M6P 2000 M6P 125 M6P 1750 KDa M 100 More NAGA in cell GLA activity (nmol/h/mL) GLA activ
Slide 9: Glycosylation pattern of in vitro expressed modNAGA
Glycosylation pattern of in vitro expressed modNAGA indicates presence of high mannose oligosacharides PNGaseF modNAGA • Cleaves high mannose, hybrid and complex oligosacharides KDa M - PNGase endoH EndoH: • Is unable to cleave N-linked complex type glycans 50 Sup 37 50 37 Cells EndoH PNGaseF In vit
Slide 10: GLA activity and reduction of LysoGb3 in Fabry mice plasma
GLA activity and reduction of LysoGb3 in Fabry (GLA-KO) mice plasma following AAV5-modNAGA injection Collaboration with Hans Aerts, Leiden GLA KO mice (n=4) IV injected with and Carlie de Vries, Amsterdam 5e13 gc/kg AAV5-modNAGA /sac. 0 2 4 6 8 10 12 wks AAV-vector GLA-activity LysoGb3 levels 2 to 1
Slide 11: LysoGb3 reduction in target organs in GLA-KO mice
LysoGb3 reduction in target organs in GLA-KO mice upon AAV-injection Collaboration with Hans Aerts, Leiden GLA-KO mice IV injected with 5e13 gc/kg AAV5-modNAGA lysoGb3 levels in organs AAV-vector liver kidney heart 60 15 6 lysoGb3 (pmol/mg protein) lysoGb3 (pmol/mg protein) lysoGb3 (pmol/mg protein)
Slide 12: ModNAGA poses a low predicted immunogenicity risk versus endogenous NAGA
ModNAGA poses a low predicted immunogenicity risk versus endogenous NAGA Collaboration with Abzena and Pro-immune Immunogenicity evaluation of modNAGA • Step 1 – In silico • Step 2 – In vitro • Algorithms to screen potential T-cell epitopes • 2 MHC class I peptides tested to common • Identify linear
Slide 13: Conclusions and future plans
Conclusions and future plans Conclusions Future plans • AAV5-NAGA results in therapeutic GLA activity and Lyso-Gb3 • Determine GLA activity and Lyso-Gb3 reduction in target reduction in plasma and target organs at already 12wks organs of AAV-injected Fabry mice (> 6 mo) • Plasma GLA activity is not
Slide 14: Acknowledgements
Acknowledgements • Chi-Lin Kuo • Linda Tan • Maria J. Ferraz • Aurelia Gondrand • Johannes M.F.G. Aerts • Dhivina Gagoomal • Carlie J.M. de Vries • Emily Mallet • Roelof Ottenhoff • H. Fogg • Jan Aten • Histoshi Sakuraba • Juan Manuel Iglesias • Ryoko Tsukahara • Michael Roberts Research Immunology
Slide 15: uniQure
uniQure
Slide 16: GLA and NAGA protein expression
GLA and NAGA protein expression GLA NAGA DELIVERING GENE THERAPY TO PATIENTS F a b r y p r o g r a m | 16
Slide 17: GLA-activity in plasma of wild type mice injected with AAV5-mod-NAGA
GLA-activity in plasma of wild type mice injected with AAV5-mod-NAGA Wild type mice IV injected with /sac. 5e13 gc/kg AAV5-modNAGA 0 2 4 7 wks AAV-vector Vector DNA GLA-activity liver plasma 10 7 120 GLA-activity (nmol/h/mL) Vehicle Genome copies /g DNA Vehicle 10 6 90 AAV5-C7-NAGA AAV5-L1-NAGA 10
Slide 18: GLA-activity in the liver and target organs of wild type animals
GLA-activity in the liver and target organs of wild type animals injected with AAV5-modNAGA Wild type mice IV injected with 5e13 gc/kg AAV5-modNAGA GLA-activity in organs AAV-vector liver kidney heart 1500 120 10 Vehicle Vehicle Vehicle (nmol/h/mg protein) (nmol/h/mg protein) (nmol/h/mg protein) AAV
Slide 19: GLA activity in liver of Fabry mice upon AAV5-modNAGA injection
GLA activity in liver of Fabry (GLA-KO) mice upon AAV5- modNAGA injection GLA-KO mice IV injected with 5e13 gc/kg AAV5-modNAGA GLA-activity AAV-vector liver 2500 GLA-activity (nmol/h/mL) 2000 Wt Vehicle Fabry Vehicle 1500 AAV5-L1-NAGA 1000 AAV5-L1-coNAGA 500 AAV5-C1-NAGA 90 AAV5-C1-coNAGA 60 30 0 12
Slide 20: Vector DNA and NAGA mRNA expression in target organs of Fabry mice
Vector DNA and NAGA mRNA expression in target organs of Fabry (GLA-KO) mice upon AAV5-modNAGA injection liver kidney heart 10 7 10 7 10 7 Genome copies /g DNA Genome copies /g DNA Genome copies /g DNA Wt Vehicle Wt Vehicle Wt Vehicle 10 6 10 6 Fabry Vehicle 10 6 Fabry Vehicle Fabry Vehicle 10 5 A