Investor and Analyst Breakfast

Transcription

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2 Investor and Analyst Breakfast American Society for Gene & Cell Therapy Annual Meeting Washington, D.C. May 12, 2017

3 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, potential, predict, project, should, "will, would and similar expressions. Forward-looking statements are based on management's beliefs and assumptions and on information available to management only as of the date of this press release. These forward-looking statements include, but are not limited to, statements regarding the development of our gene therapies, the success of our collaborations, and the risk of cessation, delay or lack of success of any of our ongoing or planned clinical studies and/or development of our product candidates. Our actual results could differ materially from those anticipated in these forward-looking statements for many reasons, including, without limitation, risks associated with collaboration arrangements, our and our collaborators clinical development activities, regulatory oversight, product commercialization and intellectual property claims, as well as the risks, uncertainties and other factors described under the heading Risk Factors in uniqure s 2016 Annual Report on Form 10-K filed with the Securities and Exchange Commission on March 15, Given these risks, uncertainties and other factors, you should not place undue reliance on these forward-looking statements, and we assume no obligation to update these forward-looking statements, even if new information becomes available in the future.

4 Welcome Matt Kapusta Chief Executive Officer Scientific Overview Harald Petry, Ph.D. Chief Scientific Officer AAV5-miHTT gene therapy for Huntington s Disease Neutralizing antibodies on efficacy of AAV delivery Repeated gene delivery in NHP with AAV5 through immune adsorption Detection of AAV vector DNA and transgene RNA in liver tissue by FISH Questions and Discussion Group Pavlina Konstantinova, Ph.D. Director, New Therapeutic Target Discovery Harald Petry, Ph.D. Chief Scientific Officer Valerie Sier-Ferreira, Ph.D. Head of Immunology Valerie Sier-Ferreira, Ph.D. Head of Immunology M A Y 1 2,

5 Circulating Anti-AAV5 Neutralizing Antibody Titers up to 1:1031 Do Not Affect Liver Transduction Efficacy of AAV5 Vectors in Non-Human Primates (Poster 198). Successful Repeated Hepatic Gene Delivery in Non-Human Primates Achieved with AAV5 by Use of Immune Adsorption (Poster 395). AAV5-miHTT Gene Therapy Demonstrates Broad Vector Distribution and Strong Mutant Huntingtin Lowering in a Huntington s Disease Minipig Model. (oral presentation) Detection of AAV Vector DNA and Transgene RNA in Liver Tissue by Fluorescent In Situ Hybridization (Poster 567). Novel AAV Vector Reservoirs: peripheral Blood Cells and Hematopoietic Progenitors. (collaborator presentation) M A Y 1 2,

6 AAV5-miHTT gene therapy for Huntington s Disease Pavlina Konstantinova, Ph.D. Director, New Therapeutic Target Discovery

7 Early HD control Neurodegenerative disorder Autosomal dominantly inherited Prevalence: 1:10,000-30,000 Age of onset around midlife 40 CAG repeat HTT DNA Prolonged CAG repeat exon 1 HTT mrna Sagittal MRI Symptoms: Motor problems/chorea Expanded polyglutamine (polyq) tract Cognitive decline Psychiatric disturbances Protein aggregation Genetic testing available Only palliative treatment Neuronal degeneration TRACK-HD M A Y 1 2,

8 Early HD control Neurodegenerative disorder Autosomal dominantly inherited Prevalence: 1:10,000-30,000 Age of onset around midlife 40 CAG repeat HTT DNA Prolonged CAG repeat exon 1 HTT mrna Sagittal MRI Symptoms: Motor problems/chorea Expanded polyglutamine (polyq) tract Cognitive decline Psychiatric disturbances Protein aggregation Genetic testing available Only palliative treatment Neuronal degeneration TRACK-HD M A Y 1 2,

9 function (%) Motor diagnosis 100 Premanifest AMT-130 Manifest slowdown disease progression presymptomatic prodromal early moderate advanced 45 Age 65 Adapted from Ross et al., Nat. Rev. Neurol. 10, (2014) M A Y 1 2,

10 ITR CAG promotor mihtt-451 polya ITR AAV5-miHTT (AMT-130): Replication deficient Adeno-associated virus, serotype 5 Designed to deliver engineered mihtt Reduction of huntingtin expression Low potential off-target effects M A Y 1 2,

11 H T T p r o t e i n l e v e l ( % ) AAV5-miHTT Striatum Cortex gc/mouse AAV5-miHTT gc/mouse AAV5-miHTT S t r i a t u m C o r t e x H T T p r o t e i n l e v e l ( % ) * * * * P B S + 5 % S u c r o s e 5. 2 x x g c / m o u s e A A V 5 - m i H T T 1. 3 x * * Control AAV5-GFP injection m u t a n t w i l d - t y p e m u t a n t w i l d - t y p e m u t a n t w i l d - t y p e H T T a l l e l e m u t a n t w i l d - t y p e m u t a n t w i l d - t y p e m u t a n t w i l d - t y p e m u t a n t w i l d - t y p e H T T a l l e l e m u t a n t w i l d - t y p e Miniarikova et al., Molecular Therapy NA 2016, Samaranch et al., Gene Therapy 2017 M A Y 1 2,

12 H T T a g g r e g a t e s D A R P P l e s i o n ( m m 3 ) Suppression of mutant huntingtin aggregation Prevention of neuronal dysfunction PBS+ 5%sucrose AAV5-miHTT PBS+ 5%sucrose AAV5-miHTT * * * * * 2. 0 * * * * * * P B S x g c 6. 5 x g c 0. 0 P B S x g c 6. 5 x g c 5 % s u c r o s e A A V 5 - G F P A A V 5 - m i H T T 5 % S u c r o s e A A V 5 - G F P A A V 5 - m i H T T Miniarikova et al., Gene Therapy, accepted M A Y 1 2,

13 1. PBS + 5%sucrose 2. 1x10 13 gc AAV5-miHTT 3. 3x10 13 gc AAV5-miHTT 4. 1x10 13 gc AAV5-GFP: thalamus striatum * * striatum thalamus * * * * * * GFP * * GFP Evers MM, ASGCT 2017 presentation 536 M A Y 1 2,

14 V e c t o r g e n o m e c o p i e s p e r g D N A M a t u r e m i H T T m o l e c u l e s / c e l l P B S + 5 % S u c r o s e 1 x g c A A V 5 - m i H T T vector DNA 3 x g c A A V 5 - m i H T T microrna L L O D P u t a m e n C a u d a t e T h a l a m u s C o r t e x 0. 1 P u t a m e n C a u d a t e T h a l a m u s C o r t e x M A Y 1 2,

15 h H T T m R N A e x p r e s s i o n ( % ) ( n o r m a l i z e d b y s s G A P D H, r e l a t i v e t o c o n t r o l ) m u t a n t h u n t i n g t i n p r o t e i n ( % ) ( r e l a t i v e t o P B S + 5 % s u c r o s e ) m u t a n t H T T p r o t e i n ( % ) ( r e l a t i v e t o P B S + 5 % s u c r o s e ) mutant HTT mrna mutant huntingtin protein * * * * * * * * * * * * s C o r t e x P B S + 5 % S u c r o s e 1 x g c A A V 5 - m i H T T 3 x g c A A V 5 - m i H T T * 5 0 * * P u t a m e n C a u d a t e T h a l a m u s C o r t e x P B S + 5 % S u c r o s e 1 x g c A A V 5 - m i H T T 3 x g c A A V 5 - m i H T T * * * * * * * 0 0 P u t a m e n C a u d a t e T h a l a m u s C o r t e x P u t a m e n C a u d a t e T h a l a m u s C o r t e x P B S + 5 % S u c r o s e 1 x g c A A V 5 - m i H T T 3 x g c A A V 5 - m i H T T P B S + 5 % S u c r o s e 1 x g c A A V 5 - m i H T T 3 x g c A A V 5 - m i H T T M A Y 1 2,

16 Dose-dependent reduction of HTT in HD rodent and tghd minipig models translates in therapeutic benefit. Widespread vector distribution upon (MRI-guided) CED delivery in NHP and tghd minipigs supported selection of striatum as the target brain structure. Long-term expression, tolerability and efficacy supports further clinical development of HTT-lowering gene therapy for HD with AMT-130. M A Y 1 2,

17 Effective liver-directed gene delivery, despite the presence of neutralizing antibodies in non-human primates Harald Petry, Ph.D. Chief Scientific Officer M A Y 1 2,

18 Impact of neutralizing antibodies (NAB) directed against AAV5 on efficacy of liver directed gene delivery in non-human primates (NHP) Approach: NAB impact on AAV5 transduction was tested in 14 NHP Sera of those 14 NHP all had pre-existing anti-aav5 NAB titers ranging from 1:57 to 1:1031 Those 14 NHP were injected intravenously with increasing doses of AAV5-hFIX: 5e11 gc/kg (n=3) 5e12 gc/kg (n=5) 2.5e13 gc/kg (n=3) 9.3e13 gc/kg (n=3) Transduction efficiency was assessed by measuring: Circulating FIX protein levels in plasma 7 days after vector infusion. Vector DNA in the liver 6 months after vector infusion (post mortem). M A Y 1 2,

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21 Demonstration that successful AAV5-based liver-directed gene delivery can be achieved in NHP, despite the presence anti-aav NAB titers up to at least 1:1031. Poses question whether patients with pre-existing anti-aav5 antibodies could benefit from AAV5-based gene therapy. M A Y 1 2,

22 Successful repeated hepatic gene delivery in non-human primates achieved with AAV5 by use of immune adsorption Valerie Sier-Ferreira, Ph.D. Head of Immunology M A Y 1 2,

23 Background: Presence of circulating neutralizing antibodies (NABs) against AAV vector capsids impair transduction of the target cells and therapeutic efficacy. Goal: To overcome anti-aav pre-existing antibodies due to exposure to wild type AAV o To increase the number of patients eligible for the therapy To overcome anti-aav antibodies raised after exposure to AAV therapy o To facilitate re-administration of AAV gene therapy Approach: Immuno-adsorption procedure M A Y 1 2,

24 Experimental set-up proof of concept in NHPs Mean Reduction levels NABs by immuno-adsorption: 11 times M A Y 1 2,

25 SEAP and hfix transgenes expression o Proteins levels o mrnas levels M A Y 1 2,

26 Data demonstrate that the use of an immune adsorption procedure enables successful re-administration of an AAV5-based gene transfer in NHPs. M A Y 1 2,

27 Fluorescent in situ hybridization (FISH): A powerful method to determine DNA/RNA distribution following AAV-based gene delivery M A Y 1 2,

28 Why FISH and IHC? To determine distribution To determine the cell specificity To quantify on the level of a cell Following liver targeted AAV gene delivery haat-gfp vector DNA/RNA is visualized in green (FISH), Albumin RNA in purple (FISH), GS protein in blue (IHC) M A Y 1 2,

29 FISH Image analysis (confocal microscopy, HALO program) IHC M A Y 1 2,

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31 The combination of FISH and IHC permit to assess the physiological transduction profile of AAV in the liver which is a valuable tool to further optimize AAV-targeting. To develop AAV-based gene therapies with increased efficiency and selectivity. M A Y 1 2,

32 Closing Remarks and Discussion M A Y 1 2,

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