Myocyte Approaches to MEA & VSO Pilot Study & Preliminary Data

Transcription

1 Myocyte Approaches to MEA & VSO Pilot Study & Preliminary Data Jiwen Zhang, Gary Gintant, Jennifer Pierson CiPA Myocyte Work Stream, Micro-Electrode Array (MEA) Subteam Voltage Sensitive Optical Probes (VSO) Subteam CSRC/HESI/SPS/FDA Meeting December 11, 2014

2 Outline Myocyte work stream: progresses and next steps Preliminary data from the pilot study MEA study data Jim Ross, MEA subteam cochair VSO study data Leslie Tung, VSO subteam cochair Acknowledgement 2

3 Comprehensive In Vitro Proarrhythmia Assay: Three Core Pillars Drug Effects on Multiple Human Cardiac Currents In Silico Reconstruction Human Ventricular Cellular Electrophysiology In Vitro Effects Human Stem- Cell Derived Ventricular Myocytes Evaluation of Clinical Drugs for Proarrhythmic TdP Liability High Risk Intermediate Risk Low Risk Preclinical ECG & Phase 1 ECG Studies: Complementary Data

4 Voltage-Sensitive Optical Probes (vso) Micro-Electrode Array (MEA) Core Component III: In vitro Effects, Human Stem Cell-Derived Ventricular Cardiomyocytes Myocyte Group (HESI): Verification of in silico reconstructions with well characterized human stem-cell cardiomyocytes Inform on repolarization effects not anticipated from ion channel and in silico reconstruction efforts

5 So Where are We and What is Next? Sept CIPA Steering Team and Work Stream Formation Nov Myocyte Subteam Formation Aug. Sept Oct. Dec Pilot Study Design and site recruitment Pilot Study Conducted Dec Feb Jan. Mar Pilot Study Data Analysis Design Validation Study Mar. Jun Jun. Sept Initiate Validation Study Complete Validation Study and Data Analysis 2013 Q3 Q4 Q1 Q2 Q3 Q4 Q1 Q2 Q CIPA Kickoff Meeting 7/23/2013 Subteam face-toface meetings and platform selection 3/11/2014 Report Pilot Study Findings 3/2/2015 Myocyte Pilot Study Completed with 8 compound blinded dataset 12/19/2014 Report Validation Study Findings 9/30/2015

6 Myocyte Pilot Study

7 Core Protocol Develop a standardized protocol including key experimental conditions & key parameters to support pilot and validation studies Provide a technical framework to facilitate Comparisons of potential proarrhythmia activity across studies: from different labs, cell types, and platforms (MEA and VSO) Evolution of myocytes, platforms, and development of new ones Core protocol components Preparation of ipsc/hesc-derived cardiomyocytes Assay conditions Compound testing for acute/direct studies Addition of drug/vehicle Assay design for multiple drug concentrations Number and type of controls End-points Quality controls Data reporting recommendations 7

8 Pilot Study Objectives and Design Objectives: Proof of concept to demonstrate MEA and VSO utility, predictiveness of proa risks using selective metrics Explore and evaluate reproducibility, variability across cell types, study sites, and platforms Investigate quality control measures to optimize assay methods Design: 4 calibration compounds; 4 test compounds; 4 concentrations, 3 triplicates Volunteer sites chose cell types and platforms Calibration Compounds Test Compounds Mexiletine (INa) Flecainide Nifedipine (ical) Moxifloxacin E-4031 (ikr) Quinidine JNJ303 (iks) Ranolazine Compound selection committee: Red: high risk; Green: low risk Yellow: not categorized or ion channel specific 8

9 Participating Sites/Cells/Platforms THANK YOU ALL! ACEA Biosciences Axiogenesis Axion Bristol-Myers Squibb Co. Cellular Dynamics ChanTest Clyde Biosciences Cyprotex GE Healthcare Janssen (JNJ) Johns Hopkins Univ. Merck NCI - Frederick Nat'n'l Lab Cancer Research NMI-TT GmbH Q-State Biosciences Sanofi Stanford Vala Sciences MEA: 12 sites, 3 platforms, 4 cell types; VSO: 4 sites; 4 platforms, 3 cell types As of Dec 3 rd, 12 (8 MEA + 4 VSO) study sites submitted 8 compound blinded datasets

10 Next Steps Wrap up pilot study by 1Q 2015 Collect remaining data from sites, perform data analysis Descriptive and statistical: site comparisons, cell-cell comparisons, variability; concentration-response Webinars/possible face-to-face meeting of VSO and MEA groups Discussions with myocyte group members and Steering Committee: Lessons learned, accomplishments, limitations of approach Draft manuscript for publication Discussions with Steering Team : Regulatory requirements Validation study expectations Plan and initiate validation study by 2Q 2015 Complete validation study and data analysis by 4Q

11 MEA Pilot Study Preliminary Data 11

12 Literature Data Demonstrating MEA s Utility and Predictiveness of ProA Risks With a myocyte assay, complexity is the advantage, assuming it can be managed so we turned to the depth of the Cardiomyocyte-MEA literature to support the protocol CM-MEA publications per year Recent work demonstrates methods to achieve both clinical predictivity and mechanism-of-action identification (Clements and Thomas, 2014). Data cut-off, Feb The MEA team standardized key experimental methods, end-point quantification, and data reporting, aided by the recently published Japanese cross-site reliability study (Nakamura et al, 2014). Pilot study: POC on metrics, evaluate variability, optimize assay methods. 12

13 Pilot Study: Four Minimum End-points Recommended definitions were provided for each end point to facilitate crosssite & cross-platform comparisons Required End Points Beat Period (s) : Max Slope-to-Max Slope Amplitude (µv): Peak-to-Peak (µv) Field Potential Duration (s): T-peak Max Slope Arrhythmia Occurrence (# of Wells) Nakamura 2014 Harris 2013 Elective endpoints, from conduction contours to contractility, were encouraged to provide additional opportunities for assay exploration and refinement 13

14 MEA Study Participants Cell Suppliers MEA Suppliers 14

15 15 Pilot Study: MEA Raw endpoints are reported for each well Site A Cell 1 Plate 1

16 Site A Cell 1 Plate 1 Pilot Study: MEA Replicate trends are computed for each compound 16

17 Sites A & B Cell 1 Plate 1 Pilot Study: MEA Trends are compared across sites for the same cell type 17

18 Sites A & B Cells 1 & 2 Pilot Study: MEA Trends are compared across 4 sites, 3 compounds, & 2 cells 18

19 Sites A & B Cells 1 & 2 Pilot Study: MEA Trends are compared across 4 sites, 3 compounds, & 2 cells 19

20 Cross-site Comparison Compound Green for 8 Sites, 4 Cells, & 3 Platforms 20

21 VSO Study Preliminary Data 21

22 Voltage Sensitive Optical Probes (VSO) Sub-group: Cell providers: Assay companies/groups: B I O S C I E N C E S Voltage sensor: Di-4-ANEPPS (small molecule) QuasAr2 (protein) FluoVolt (small molecule) Di-4-ANEPPS (small molecule) Detector: Photomultiplier CMOS camera CMOS camera CMOS camera

23 Action potential shape differs across the different cell providers 200 ms 200 ms 200 ms % Repolarization APD (ms) Axiogenesis CDI GE-Healthcare APD30 APD50 mean+sd APD90 Depolarization (Upstroke) Na + channels 100 ms Repolarization phase LTCC herg channel 23

24 24 VSO end-points: Minimum end-points include measures of repolarization timings and arrhythmia detection. Additional parameters are encouraged to facilitate mechanism identification. Primary end points: Spontaneous beat rate and regularity Action potential rise time (10-90%) APD at 30%, 50% & 90% (APD30, APD50, APD90) AP triangulation index (=APD90-APD30) Incidence of early after depolarisations (EADs) Rate sensitivity of the above parameters (paced only) Baseline Drug 200ms

25 Pilot Study: VSO Effects are shown here at 1 site of Compound Red effects on APD30, APD90, rise time and triangulation. Baseline Drug Baseline Drug Baseline Drug Baseline Drug 200ms 200ms [Drug] C1 C2 C3 C4 APD 30 (% of baseline) TRise (% of baseline) Vehicle (DMSO) Drug C1 C2 C3 C4 [Drug] Vehicle (DMSO) Drug C1 C2 C3 C4 [Drug] APD 90 (% of baseline) Triangularization (% of baseline) ms Vehicle (DMSO) Drug * 200ms *** C1 C2 C3 C4 [Drug] Vehicle (DMSO) Drug * C1 C2 C3 C4 [Drug] 25

26 Pilot Study: VSO Trends are compared here across all 4 sites for Compound Red effects on APD90. Compound Red 26

27 Cell 1 Cell 1 Pilot Study: VSO Trends are compared here across 2 sites, 3 compounds, & 2 cells Compound Green Compound Red Compound Blue Concentration Concentration Concentration 27

28 Acknowledgement AbbVie ACEA Biosciences Axiogenesis Axion Bristol-Myers Squibb Co. Cellular Dynamics ChanTest Clyde Biosciences Cyprotex FDA GE Healthcare HESI Janssen (JNJ) Johns Hopkins Univ. Merck NCI - Frederick Nat'n'l Lab Cancer Research NMI-TT GmbH Q-State Biosciences Sanofi Stanford Vala Sciences Gary Gintant Leyna Zhao, Xiaoyu Zhang, Yama Abassi Greg Luerman, Brian Murphy, Ralph Kettenhoffen Jim Ross, Daniel Millard Hong Shi, Paul Levesque Arne Thompson, Blake Anson Jessica Brimbecombe Godfrey Smith, Margaret Anne Craig Chris Strock Qianyu Dang Jan Turner, Michael Clements, Jiwen Zhang Jennifer Pierson, Kyle Brunette Hua Rong Lu Les Tung, Renjun Zhu, Venkatesh Hariharan John Imredy Frederick Sannajust Liang Guo Udo Kraushaar, Guenther Graham Dempsey, Celine Hechard Jean-Michel Guillon Joe Wu, Yingxin Li, Priyanka Garg Ross Whittaker 28

29 Appendix 29