uniQure N.V. · NASDAQ:QURE
uniQure — HD Therapeutics Conference 2020
43 slides from uniQure's conference presentation, Feb 27, 2020
Slide 1: Translatable biomarkers in gene therapy for Huntington Disease: learnings from pre-clinical studies
Translatable biomarkers in gene therapy for Huntington Disease: learnings from pre-clinical studies Astrid Vallès-Sánchez, PhD Senior Scientist, Biomarker Lead
Slide 2: Forward-looking Statements
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
Slide 3: A pioneer in gene therapy
A pioneer in gene therapy +20 years of experience in AAV gene therapy Research & Development Development and GMP Manufacturing Amsterdam, The Netherlands Lexington (MA), USA A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 3
Slide 4: Pipeline
Pipeline A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 4
Slide 5: First AMT-130 clinical study in HD patients
First AMT-130 clinical study in HD patients (HTT-lowering gene therapy) • Phase 1/2a clinical study: • Objective is to test safety of AMT-130 • This will be tested in adult early manifest HD patients • The first study will take place in the US • Currently in phase of site activation and patient recr
Slide 6: What is AMT-130 and how do we deliver it?
What is AMT-130 and how do we deliver it? AAV5-miHTT (Company name AMT-130) One-time intracranial injection of AMT-130 into striatum (caudate nucleus and putamen) AAV5 capsid Expression Caudate nucleus cassette Putamen ITR ITR miR-451-HTT CAG promoter polyA • Replication deficient AAV5 • Targeting t
Slide 7: Proof-of-Concept of AMT-130 gene therapy: from patient cells to large animals
Proof-of-Concept of AMT-130 gene therapy From patient cells to large animals HD Rat2 Non-human primate1 4 types HD Mouse4,6 tgHD Minipig3 Cultured human neurons5 Diseased rodent models NHPs HD minipigs Efficacy Efficacy Efficacy Safety Safety Safety Distribution 1. Samaranch L et al. Gene Ther 2017;
Slide 8: Building a biomarker panel for AMT-130
Building a biomarker panel for AMT-130 PK PD Disease modification Clinical outcome Pharmacokinetics Pharmacodynamics Understanding of biological mechanism Drug exposure What the drug Effects on key Therapeutic levels does to the body aspects of disease clinical benefit • Translational PK/PD measures
Slide 9: PK and PD measures: vector DNA, miHTT, HTT
ITR ITR miR-451-HTT CAG promoter polyA PK: vector DNA miHTT PD: HTT mRNA HTT protein Downstream disease markers A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 9
Slide 10: Biomarker panel AMT-130
Biomarker panel AMT-130 Imaging biomarkers Volumetric MRI miHTT Striatal MRS HTT protein (mutant and total) NFL CSF biomarkers A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 10
Slide 11: Evaluating miHTT
Evaluating miHTT HD neurons NHP tgHD minipig miHTT CSF biomarkers A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 11
Slide 12: Measuring miHTT in extracellular vesicles
Measuring miHTT in extracellular vesicles EVs are an important source of extracellular miRNAs Sorting of miRNAs into EVs is a regulated process EVs are present in all body fluids including CSF → miHTT in CSF EVs? M. Sogorb-Gonzalez (Poster nr. 83) A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONF
Slide 13: EV-miHTT is released from AMT-130 treated HD neurons in a dose-dependent manner
EV-miHTT is released from AMT-130 treated HD neurons in a dose-dependent manner 10 7 10 6 DAY 12 miHTT molecules/ 10mL medium 10 5 ExoQuick-TC™ 10 4 10 3 LLOD 10 2 10 1 Confirmed EV identity with Western Blot, 0 105 106 107 FACS analysis, TEM and SEC EV isolation MOI AAV5-miHTT M. Sogorb-Gonzalez (P
Slide 14: Robust correlation between viral dose, cellular miHTT and secreted EV-miHTT
Robust correlation between viral dose, cellular miHTT expression and secreted EV-miHTT Viraldose Viral dose Cellular miHTT Cellular miHTT vs vs. vs vs. EV-miHTT EV-miHTT EV-miHTT EV-miHTT 1.1 1.1 EV-miHTT detection EV-miHTT detection 1.0 1.0 0.9 0.9 0.8 0.8 R2=0.8786 R2=0.9431 p=0.006 p=0.001 0.7 0.
Slide 15: Can EV-miHTT be detected in CSF?
Can EV-miHTT be detected in CSF? • Assessment in NHP study, MRI-guided CED of AMT-130 (caudate and putamen) Striatum Thalamus, Peripheral Cortex Midbrain organs Low dose 10 7 High dose (molecules/μg RNA) 10 6 miHTT 10 5 10 4 10 3 Sp r en y K n id s ita e St be Th tum e ra e ve ne u b rie lob po lob
Slide 16: EV-miHTT is stably expressed in NHP CSF after intrastriatal delivery
EV-miHTT is stably expressed in NHP CSF up to 6 months after intrastriatal delivery of AMT-130 CSF Vector DNA CSF miHTT molecules 10 11 10 6 10 5 (molecules /mL) 10 9 Vector DNA CSF miHTT 10 4 (gc/ml) 10 7 10 3 10 2 10 5 10 1 LOD 10 3 10 0 0 4 8 12 16 20 24 0 4 8 12 16 20 24 † Weeks Post-dose Weeks
Slide 17: CSF miHTT beyond months post-administration?
CSF miHTT beyond 6 months post-administration? • Stable miHTT expression in CSF of tgHD minipigs: up to 2 years (ongoing study) 10 6 Stable expression 10 5 (molecules/mL CSF) CSF miHTT 10 4 10 3 10 2 10 1 o o o e o o o o o os 3m 6m 9m m m m m m 12 15 18 21 24 d e- pr C. Vendrell-Tornero A. Vallès, M
Slide 18: CSF miHTT as PK measure
CSF miHTT as PK measure HD neurons NHP tgHD minipig miHTT EV-miHTT can reliably be measured in CSF and used as a translational measure of gene CSF biomarkers therapy persistence A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 18
Slide 19: Evaluating CSF HTT protein and CSF NFL
Evaluating CSF HTT protein and CSF NFL tgHD minipig HD mice HTT protein (mutant and total) NFL CSF biomarkers A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 19
Slide 20: Long-term study in tgHD minipigs
Long-term study in tgHD minipigs Ubiquitous expression of human mutant HTT (548aa N-term fragment) Good model to assess biodistribution and target engagement in large brain tgHD minipig model: PIGMOD Center and CHDI A. Vallès, M. Evers et al. (submitted) A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTI
Slide 21: Assessment of AMT-130 biodistribution and efficacy
Assessment of AMT-130 biodistribution and efficacy Extensive brain dissection and bioanalysis sagittal view 6 months interim sacrifice → n=54 punches (alternate slices) 12 months interim sacrifice → n=170 punches (all slices) Bioanalysis • DNA isolation → vector DNA (Q-PCR) • RNA isolation → miHTT (
Slide 22: Vector DNA distribution in tgHD minipig brains shows extensive coverage
vector DNA distribution in tgHD minipig brains shows extensive brain coverage comparable at 6 and 12 months A Cortex Striatum 6 months B C 10 8 12 months 10 7 Cortex (12 months) 10 7 Spinal cord (12 months) 10 7 VG copies / μg DNA VG copies / μg DNA VG copies / g DNA 10 6 10 6 10 6 10 5 10 5 10 5 1
Slide 23: Strong long-term lowering of mutant HTT protein in tgHD minipig brain
Strong long-term lowering of mutant HTT protein in the tgHD minipig brain, comparable at 6 and 12 months A. Vallès, M. Evers et al. (submitted) A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 23
Slide 24: Mutant HTT protein in CSF is lowered, but does not reflect brain lowering
Mutant HTT protein in CSF is also lowered, but does not accurately reflect mutant HTT lowering in brain tissue Increase in CSF mHTT levels with age Control (naive) 400 rAAV5-miHTT CSF mtHTT (fM) 300 200 injection 100 0 3 6 9 12 15 18 tgHD minipig age (months) AMT-130 datapoints from 3-12 months post
Slide 25: Mutant HTT protein in CSF is lowered, but does not reflect brain lowering
Mutant HTT protein in CSF is also lowered, but does not accurately reflect mutant HTT lowering in brain tissue Brain mHTT protein lowering Increase in CSF mHTT levels with age 40% 45% → Prefrontal cortex Control (naive) 80% 75% → Caudate 85% 70% → Putamen rAAV5-miHTT CSF mHTT (% from control) 400 12
Slide 26: Stable mutant HTT protein lowering in CSF of tgHD minipigs
Long-term in-life monitoring demonstrates stable mutant HTT protein lowering in CSF of tgHD minipigs • Longitudinal data collected up to two years • Stable CSF mutant HTT protein lowering of on average 25-30% CSF mHTT (% from control) 125 100 75 30% 50 50% 70% 25 0 pre-dose 6m 12m 18m 24m A. Vallès,
Slide 27: Evaluation of CSF NFL: no phenotype in young tgHD minipigs
Evaluation of CSF NFL: no phenotype in (young) tgHD minipigs 1250 1250 WT CSF NF-L (pg/mL) CSF NF-L (pg/mL) 1000 1000 tgHD 750 750 500 500 250 250 0 0 WT tgHD 20 30 40 age (months) • No differences in NFL levels in WT versus tgHD minipigs (up to 4 years of age) • NFL phenotypic correction after HTT
Slide 28: CSF NFL transiently elevated after dosing, returning to baseline
CSF NFL is transiently elevated after dosing, returning to baseline and remaining stable throughout study period 50000 10000 WT Control (naive) 40000 8000 rAAV5-miHTT CSF NF-L (pg/mL) CSF NF-L (pg/mL) 30000 6000 20000 4000 10000 2000 0 0 3m 6m 9m m se T 3m 6m 9m m se T d d W 14 28 W 12 12 do do e- e
Slide 29: In-life phase allows for long-term CSF biomarker monitoring
In-life phase allows for long-term CSF biomarker monitoring miHTT mHTT protein NFL 10 6 125 80000 CSF mHTT (% from control) Control (naive) 10 5 100 rAAV5-miHTT (molecules/mL CSF) CSF NF-L (pg/mL) 60000 CSF miHTT 10 4 75 30% 40000 10 3 50 50% 70% 20000 10 2 25 10 1 0 0 3m 6m 9m e m m m m m 28d d 3m
Slide 30: What about HTT exon1?
What about HTT exon1? (CAG)n 1 2 3 4 67 HTT RNA mis-splicing 67 HTT mRNA (CAG)n (CAG)n 1 (AAAA)n 1 67 (AAAA)n Highly pathogenic Exon 1 HTT protein Full-length HTT protein Sathasivam K et al. Proc Natl Acad Sci U S A 2013; 110:2366 | Gipson TA et al. RNA Biol 2013; 10:1647 | Neueder A et al. Sci Rep
Slide 31: Full-length and exon HTT mRNA lowering in striatum and cortex
Full-length and exon 1 HTT mRNA lowering in striatum and cortex after treatment with AAV5-miHTT HD mice (homQ175-FDN) Southwell AL et al. Hum Mol Genet 2016; 25:3654 Full length HTT in StriatumFull length HTT inHTT Full-length Striatum in Cortex 1.5 1.5 1.5 VEH Relative expression Relative expressio
Slide 32: CSF HTT protein and CSF NFL as efficacy measures
CSF HTT protein and CSF NFL as efficacy measures tgHD minipig HD mice HTT protein (mutant and total) NFL Stable CSF mHTT protein lowering up to two years post-injection However: CSF mHTT underestimates level of lowering in deep brain regions NFL confirmed as safety readout Need of (protein) translat
Slide 33: Evaluating striatal MRS
Evaluating striatal MRS Imaging biomarkers Volumetric MRI Striatal MRS HD mice A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 33
Slide 34: Magnetic resonance spectroscopy (MRS)
Magnetic resonance spectroscopy (MRS) • Allows noninvasive and in vivo exploration of the molecular composition of tissue • Brain metabolites in target regions • Safety and efficacy measure Reproducible alterations of striatal MRS in HD: N-acetyl aspartate (NAA), neuronal integrity marker Myo-in
Slide 35: Striatal MRS metabolite concentrations in Q175-FDN mice reflect HD patients
Striatal MRS metabolite concentrations in Q175-FDN mice reflect findings in HD patients BR Leavitt (Poster nr. 87) Southwell AL et al. Hum Mol Genet 2016; 25:3654 | Leavitt BR et al. (in preparation) A GLOBAL LEADER IN GENE THERAPY HD THERAPEUTICS CONFERENCE – FEBRUARY 27, 2020 | 35
Slide 36: Striatal MRS metabolite concentrations in Q175-FDN mice reflect HD patients
Striatal MRS metabolite concentrations in Q175-FDN mice reflect findings in HD patients Can MRS profile be reversed by AAV5- miHTT mediated HTT lowering? BR Leavitt (Poster nr. 87) Southwell AL et al. Hum Mol Genet 2016; 25:3654 | Leavitt BR et al. (in preparation) A GLOBAL LEADER IN GENE THERAPY HD
Slide 37: Reversal of striatal MRS metabolites in homQ175FDN mice
Reversal of striatal MRS metabolites in homQ175FDN mice suggests potential to reverse HD neuropathology Neuronal integrity Glyosis 6.0 Q175FDN 7 Q175FDN Mean concentration Mean concentration 5.5 *** 6 * tNAA (mM) mI (mM) 5.0 5 4.5 4 4.0 3 WT VEH VEH LOW HIGH WT VEH VEH LOW HIGH AMT-130 AMT-130 Rever
Slide 38: MRS correction is reflected in striatal transcriptomic profile
MRS correction is reflected in striatal transcriptomic profile Heatmap clustering of top40 DE genes Neuronal integrity 6.0 Q175FDN Mean concentration 5.5 *** tNAA (mM) 5.0 4.5 4.0 WT VEH VEH LOW HIGH AMT-130 BR Leavitt (Poster nr. 87) SB Thomson, BR Leavitt et al. (in preparation) A GLOBAL LEADER IN
Slide 39: Evaluating striatal MRS
Evaluating striatal MRS Imaging biomarkers Volumetric MRI Striatal MRS Striatal MRS shows great potential to detect efficacy signals Reversal of HD-like MRS profile by HTT lowering is reflected by changes towards normalization in striatal transcriptomic signature A GLOBAL LEADER IN GENE THERAPY HD T
Slide 40: Biomarker panel to assess efficacy of AMT-130 treatment
Biomarker panel to assess efficacy of AMT-130 treatment Expectations of the response of each biomarker set based on preclinical data miHTT Biofluid HD Imaging expression biomarkers biomarkers HTT protein in CSF Striatal MRS (metabolite profile) EV-miHTT and free miHTT in CSF NFL in CSF and plasma Vo
Slide 41: AMT-130-01 Phase 1/2a study: Key endpoints
AMT-130-01 Phase 1/2a study: Key endpoints Key primary endpoints (SAFETY) Key secondary endpoints • Adverse events • Levels of vector DNA and miRNA expression in CSF over time • Clinical parameters • Laboratory parameters in blood Key exploratory endpoints and CSF • HTT protein levels in CSF • Chang
Slide 42: Thank you
THANK YOU!! Institute of Animal Physiology and Genetics (Czech Republic) Bozena Bohuslavova | Jiri Klima | Zdenka Ellederova | Jan Motlik CHDI (USA) Doug Macdonald | David Howland Amsterdam UMC (The Netherlands) Rienk Nieuwland IRBM (Italy) Roberta Pintauro | Valentina Fodale | Alberto Bresciani Uni
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