Functional Enhancement of Human iPSC-derived Cardiomyocytes Enabling Assessment of Inotropic Compounds and Improved Prediction of Compound Risk Yama Abassi, PhD Vice President of Global Strategic Business Development and Grow th ACEA Biosciences Blake Anson, PhD Global Business Director Toxicology and Safety Pharmacology September 25, 2017
Outline – Cellular Dynamics and iCell Cardiomyocytes • CDI Company Overview • iCell Cardiomyocytes Understand your model Relative to the ‘ideal’ Current utility and advantages October 2, 2017 2
The Power of IPSC technology Basic research Drug Discovery Cell Therapy Bringing relevant human biology and diversity to basic research, drug discovery, and therapy October 2, 2017 3
Company Overview • Cellular Dynamics International (CDI) is a leader in production and application of human iPS cells and iPS cell-derived cell types • Acquired by FUJIFILM (4/2015); International presence Headquartered in Madison, WI (additional site in Novato, CA) Application / Distribution sites in Japan, South Korea, and Tilburg Local Sales and FAS support • Currently employs ~175 total staff w/ >900yrs cumulative stem cell and differentiation experience • >100 patents (owned or licensed) • Portfolio includes off the shelf products, as well as, custom cell production and assay services. October 2, 2017 4
CDI: Life Sciences and Regenerative Medicine 2 Business Divisions o Life Sciences o Cellular Therapeutics Life Sciences Division serves 4 major market areas o Basic and Translational Sciences o Safety Pharmacology & Toxicity o Drug Discovery and Bioengineering o Specialty Markets Cellular Therapeutics Division has 2 focal areas o Internal cell therapy programs o Ocular, Cardiac, Neurodegenerative, and Oncology o Contract Development and Manufacturing partnerships An unyielding commitment to consistent and robust iPSC-based solutions for current and future research and therapeutic applications October 2, 2017 5
Life Science Research: Current Product Portfolio iCell iCell iCell iCell iCell DopaNeurons Motor Neurons Astrocytes Neurons GlutaNeurons iCell iCell iCell iCell Cardiac iCell Cardiomyocytes Hepatoblasts Hepatocytes Progenitor Cells Macrophages iCell iCell iCell MSCs Skeletal Myoblasts Endothelial Cells HPCs New Products in development: Early access availability iCell RPEs October 2, 2017 6
Human iPSC Models Functional recapitulation Cardiomyocytes Neurons Neurite outgrowth / retraction Electrical activity Ca 2+ handling Synaptogenesis / pruning Ion channel and synaptic activity Contractility October 2, 2017 7
iCell Cardiomyocytes; Contextual Relevance Functional and Structural toxicity Structural Toxicity Functional Toxicity - 1 o effect is on electrical/mechanical function 1 o effect is on general cellular processes Electrical Viability Ion channels, Action Potentials, GPCRs Lipid accumulation Mitochondrial function Cell Signaling • Ca 2+ signaling (EC coupling) Oxidative stress Bioenergetics • Biochemical etc….. Mechanical Contractility Contextual relevance enables both functional and structural mechanistic toxicity testing Processes are linked, thus downstream biology is a phenotypic biomarker for upstream activity October 2, 2017 8
iPSC-Cardiomyocytes moving from novelty to mainstream Pubmed results for stem+cell+cardiomyocytes+toxicity 350 Total Publications 300 Contemporary model with great interest 250 • Exponential increase in publications 200 Regulatory evaluation 150 • CiPA, JiCSA, CSA-Hi 100 Not entirely free of debate 50 • Focus here will be functionality, utility, 0 and advantages w/rspct to current models Publication year >300 publications on toxicity >5900 publications stem+cell+cardiomyocytes October 2, 2017 9
Differences Between iPSC and Adult Cardiomyocytes What is the impact of some of these differences - What is the limit of utility? Example procedures to ‘mature’ cardiomyocytes past these differences Denning et al., 2016 October 2, 2017 10
Cardiomyocyte Electrophysiology Differences between native ventricular myocytes and iPSC cardiomyocyte action potentials Differing ion channel / current stoichiometry iPSCs primarily show: • Decreased I Na • Decreased I K1 • Increased I funny • Mixture of cellular subtypes Spontaneously beating iPSC-cardiomyocytes with depolarized MDP October 2, 2017 11
iCell Cardiomyocytes and iCell Cardiomyocytes 2 Overview Disease modeling /Target ID / Toxicity Testing Human Cardiomyocytes Screening Functional and Structural Toxicity Greater predictivity Hypertrophy MOA Identification Dilated Cardiomyopathy Diabetic Cardiomyopathy Ischemia/reperfusion Guo 2011, 2013 Regenerative Medicine C. Scott Tox Sci 2014 >95% pure Regulatory Interactions Cardiac Patch / Catheter Normal human biology Delivery Predictive human reagent Gold Standard ~100 publications > 90% Top Pharma Used by International Regulatory Agencies October 2, 2017 12
iCell Cardiomyocytes Characterization Electrophysiology Ionic Currents Spontaneous Action Potentials I Na I Ca-L I to I Kr G a s – b 1 G a q – a 1 I funny I K1 G a i – m2 Carbachol Isoproterenol Phenylephrine Frequency Ma, et al, Am. J. Physiol., 2011 Concentration ( m M) Drug Control iCell Cardiomyocytes possess the appropriate ion channels, action potentials, and GPCR pathways expected of a relevant human cardiomyocyte model October 2, 2017 CONFIDENTIAL 13
Value Proposition Physiologically Appropriate Arrhythmia Triggers Adult Canine Purkinje Fiber APs iCell Cardiomyocyte APs -1.87±0.26 mV/mV -2.28±0.11 mV/mV January et al, Circ Res , 65:570+, 1988 Ma et al, AJP:H&C , 301:H2006+, 2011 iCell Cardiomyocytes show physiologically relevant proarrhythmic triggers October 2, 2017 CONFIDENTIAL 14
Toxicity Testing Predictivity Screens Proarrhythmia screening in 96 wells Larger screens with quantitative analytics provides greater predictivity • >120 compounds • ~equal positives and negatives • beat rate, atypical beats, irregularity Cardiomyocyte activity generates rhythmic deflections of the impedance baseline Guo et al., 2013 > 90% -- QT prolongation > 80% -- Proarrhythmia Guo et al., 2011 iCell Cardiomyocytes provide a more Easily implemented higher throughput predictive tool for detecting proarrhythmia proarrhythmia screening October 2, 2017 15
iPSC Cardiomyocytes Contractility iPSC Cardiomyocytes Adult Cardiomyocytes iPSC versus adult cardiomyocytes • Isotropic myofilament arrangement • Isotropic cell alignment • Negative force frequency relationship • Limited effects of pre-load (Frank-Starling) • Difficult to directly translate positive inotropy October 2, 2017 16
Measuring Contractility Comparison to Gold Standard Comparisons between IonOptix-based measurements of dog cardiomyocytes (gold standard) to Ca 2+ and impedance-based measurements of iCell Cardiomyocytes (higher throughput) C. Scott Tox Sci 2014 October 2, 2017 17
Measuring Contractility Comparison to Gold Standard Comparisons between IonOptix-based measurements of dog cardiomyocytes (gold standard) to Ca 2+ and impedance-based measurements of iCell Cardiomyocytes (higher throughput) Dog iCell Cardiomyocytes cardiomyocytes IonOptix 1 FLIPR 2 Parameter Impedance 3 83% 77% sensitivity 90% 84% 70% specificity 74% 82% 74% accuracy 84% 90% 79% pos predict 85% 76% 67% neg predict 82% C. Scott Tox Sci 2014 1 AR Harmer Tox App Pharm (2012) 2 , A. Pointon Tox Sci (2014) 3 , C. Scott Tox. Sci (2014) iCell Cardiomyocytes • Show potency correlation with gold standard model • Demonstrate good to excellent assay validation parameters • provide a predictive surrogate model for measuring contractility October 2, 2017 18
Small molecule KI-induced cardiotoxicity Phenotypic Assays Prediction hindered by : FDA approved SMKI show cardiac liabilities • • Highly conserved site of action-ATP-binding pocket Preclinical assays were insufficient • (on vs off target effects) Toxicities arose in late development / clinic • • Multiple effects on overlapping endpoints Difficult to ascribe mechanism Cellular impedance assays with iCell CMs can predict KI toxicity Altered beat phenotype indicates upstream interaction S. Lamore SOT 2014 Model can: • Determine on-target vs off-target KI toxicity (MARK vs Chk KI) iCell Cardiomyocytes provide a predictive • Identify KI-related toxicity with p<0.05 tool for detecting KI toxicity (>160 cmpds via Ambit and AZ-proprietary datasets) See also Cohen et al, 2013, Doherty 2013, Talbert 2014, Peters et al, 2015 M. Peters CDI UGM 2014 Lamore et al., 2017 October 2, 2017 19
Implementing iCell Cardiomyocytes in toxicity testing cascade Structural Toxicity Cell injury Cell death Functional Toxicity Proarrhythmia Ca 2+ handling Contractility Peters et al., (2015) Cardiovasc Toxicol Primary screen identifies a problem (or lack thereof) Secondary investigations identify mechanism Subsequent primary screens can be performed on med chem series October 2, 2017 21
iCell Cardiomyoytes / Cardiomyocytes 2 Recapitulate native behavior Predictive for proarrhythmia, altered contractility, and structural toxicity Useful model for predicting adverse effects of small molecule and biologics-based therapies www.cellulardynamics.com October 2, 2017 22
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