Automated Patch Clamp User Meeting 2017 th June 9 2017 Cambridge, MA Automated Patch Clamp User Meeting 2017 Key Note Speakers Dr. Chris Miller, Professor Biochemistry Brandeis University: A Weird Ion Channel for a Very Weird Ion Dr. Owen McManus, CTO Q-State Biosciences Inc.: Phenotypic Assays of Neuronal Function to support Drug Discovery Dr. Owen McManus has extensive experience in ion channel drug discovery and technology development. He received his Ph.D. in Physiology from the University of Utah in 1983 and went on to complete his Postdoc in the lab of Dr. Karl Magleby, focusing on Physiology & Biophysics. He spent more than twenty years devoted to ion channel drug discovery efforts in pharmaceutical and academic settings for a range of therapeutic areas (pain, glaucoma, diabetes, asthma, hypertension, and immunosuppression) leading to selection of several preclinical and clinical candidate compounds. He has Dr. Chris Miller studies the molecular contributed in several areas in basic drug discovery mechanisms by which ion channels and including target identification and validation, assay and membrane transport proteins work. He technology development, lead identification and received his Ph.D. in Molecular Biology from optimization, and early clinical studies. He has worked on the University of Pennsylvania in 1974 and technology and instrument development in support of went on to complete his Postdoc in drug discovery leading to release of commercial Biochemistry at Cornell University in 1976. instruments and reagents. His recent work focuses on Dr. Miller pioneered techniques to record developing and utilizing optical technologies to enable the activity of ion channels reconstituted improved disease models, patient stratification and drug into planar phospholipid membranes allowing the study of the single-molecule discovery. behavior of these proteins in a biochemically defined system. His research career has spanned fundamental changes in the ways these membrane proteins can be studied, from cellular patch-recording and membrane reconstitution, to recombinant DNA manipulation, to high- level expression, and x-ray crystallography. He has endeavored to stay current with all these developments, bringing them to bear on questions of membrane transport mechanism. ]2[ Cambridge, Massachusetts| June 9th 2017 Table of Contents Morning Session Agenda…………………….…….……….4 Afternoon Session Agenda………………….….………….5 Poster Session Abstracts…………………….…….………...6 Event Area Map……………………….…………..….……..15 ]3[ Automated Patch Clamp User Meeting 2017 Morning Session Agenda Arrival & Breakfast 8:15 – 9:00 AM Charles View Ballroom 16th Floor Poster Set Up Foyer Area 16th Floor 9:00 – 9:15 AM Rodolfo Haedo, Senior Vice President Nanion Technologies: Welcome & Introduction 9:15 – 10:00 AM – Keynote – Dr. Chris Miller, Professor Biochemistry Brandeis University: A Weird Ion Channel for a Very Weird Ion. 10:00 – 10:30 AM Dr. Alfred George, Professor & Chair Department of Pharmacology Northwestern University: Automated Electrophysiology of iPSC-derived Cardiomyocytes. 10:30 – 11:00 AM Dr. Tim Strassmaier, Senior Scientist Nanion Technologies: Pursuit of TRP channel targets with the SyncroPatch 384PE. Poster & Coffee Break 11:00 – 11:15 AM Foyer Area 16th Floor 11:15 AM – 11:45 AM Andrew Allen & David Baez, Broad Institute: The Agony and Ecstasy: Characterization of Disease-Related Ion Channel Variants using the SyncroPatch 384PE. 11:45 AM – 12:15 PM Dr. Bjorn Knollmann, Director Vanderbilt Center for Arrhythmia Research: Modeling Heart Disease with human iPSC-derived Cardiomyocytes - First Experiences with CardioExcyte 96. Lunch 12:15 – 2:00 PM 16th floor ]4[ Cambridge, Massachusetts| June 9th 2017 Afternoon Session Agenda 1:15 – 2:00 PM – Keynote – Dr. Owen McManus, CTO Q-State Biosciences Inc.: Phenotypic Assays of Neuronal Function to support Drug Discovery. 2:00 - 2:30 PM Dr. Maria Barthmes, Product Manager Nanion Technologies: Targeting Transporters: High Throughput Screening and Characterization of Membrane Transporters and Pumps with the SURFE2R 96SE. 2:30 - 3:00 PM Dr. Matt Fuller, Senior Scientist Icagen, Inc.: Advancing Early Drug Discovery with the SyncroPatch 384PE at Icagen. 3:00 - 3:30 PM Dr. Greg Kaczorowski, CEO Kanalis Consulting, L.L.C.: Lessons Learned in Drugging Ion Channels from Prosecuting the Novel ROMK Target. 3:30 - 4:00 PM Poster and Coffee break Foyer Area 16th Floor 4:00 - 4:30 PM Dr. Srinivasan Venkatachalan, Assist Principal Scientist Chromocell Corporation: Ion Channel Therapeutic Discovery and Development Effort at Chromocell Corporation using Patchliner. 4:30 - 5:00 PM Dr. Carlos Vanoye, Research Assoc. Professor Northwestern University: High Throughput Functional Analysis of Ion Channel Gene Variants. 5:30 Reception and Dinner Foyer Area 16th Floor ]5[ Automated Patch Clamp User Meeting 2017 Poster Session Abstracts 1. Next level toxicity screening: From single channel to overall cell behavior Conrad Weichbrodt1, Mohamed Kreir1, Matthias Beckler1, Alison Obergrussberger1, Ilka Rinke1, Michael George1, Sonja Stoelze-Feix1, Andrea Brüggemann1, Niels Fertig1 1Nanion Technologies GmbH, Munich, Germany Many lead compounds fail in the late stages of the drug development process mainly by inflicting drug induced injury on liver, heart or other organs. Therefore, devices for detecting possible cell toxicity in early stages of the development process are highly desired. Ion channels represent an especially important class of drug targets for in vitro pharmacological profiling. High throughput screening (HTS) assays such as automated electrophysiological patch clamp and impedance based assays allow for the determination of drug effects on a wholecell level whereas artificial bilayers provide a robust environment for the assessment of single ion channel molecules. We here present the CardioExcyte 96, a system providing a combination of Electric Impedance Spectroscopy (EIS) as well as Electric Field Potential (EFP) readout for a network of diverse cells like iPS cardiomyocytes or hepatocyte-like cells which is exemplified by toxicity effects utilizing reference compounds such as Dofetilide (on iPS cardiomyocytes) or Paracetamol (on hepatocyte-like cells). Furthermore we present the temperature dependent activation or deactivation of different Transient Receptor Potential (TRP) channels by means of planar patch clamping on our HTS platforms Patchliner and SyncroPatch 384PE as well as with highest resolution on a single channel level on our recently introduced Orbit mini setup. 2. A new analysis pipeline to improve assessment of cardiac liability in high throughput electrophysiology screens with routine MoA detection for slow onset compounds Stephan Steigele1, Ana L. Teixeira2, Martin Ginkel1, Verity A Talbot2, Lisa J McWilliams2, Matt Bridgland-Taylor2, Stephan Heyse1 1Genedata AG, Basel, Switzerland, 2AstraZeneca, Cambridge Automated patch clamp screens generate highly valuable information for drug research programs. In safety pharmacology in particular, early testing is nowadays considered essential to manage the risk of late failure and patient health. Thus, more predictive and affordable screens need to be introduced to increase both specificity and throughput e.g. for cardiovascular liability detection and resolution. At AstraZeneca, we have implemented the Nanion SyncroPatch 384PE electrophysiology platform in standard 384-well plate format, which delivers higher throughput measurements for hERG and other key cardiac ion channels at dramatically reduced consumable costs and experiment time, whilst capturing full channel response kinetics. Jointly with Genedata ]6[ Cambridge, Massachusetts| June 9th 2017 we developed a pipeline for processing and analyzing this complex data. This pipeline reads binary raw data directly from the SyncroPatch 384PE instrument into Genedata Screener®, then assigns and integrates the sweep recordings from cumulative compound addition series, and finally subjects the recordings to scientist review that uses powerful filtering and masking based on quality control measurements. We further developed two methods: The first to normalize the cumulative curve recordings, which are compensated for signal variation using time-match control, and the second to automatically detect “slow onset” compounds using a sigmoidal fit model per concentration step. Because of their slow onset, these compounds were previously potentially underestimated, but are now flagged. This investment in a next-generation automated electrophysiology platform and an appropriate automated data processing pipeline is resulting in more predictive and affordable safety screens at high scalability and rapid turnover for discovery projects across AstraZeneca. 3. Preclinical Cytotoxicity Investigations in Stem Cells for Primary and Secondary Assays with an “All Inclusive” Approach Corina T. Bot2, Sonja Stölzle-Feix1, Nadine Becker1, Ulrich Thomas1, Krisztina Juhasz1 , Leo Doerr1, Matthias Beckler1, Claudia Haarmann1, Alison Obergrussberger1, Susanne M. Rehnelt3, Tobias Bruegmann3, Philipp Sasse3, J-F. Rolland4,5, R. Rizzetto4,5, V. Agus4,5, L. Redaelli4,5, Rodolfo J. Haedo2, Andrea Brüggemann1, Michael George1, Niels Fertig1 1Nanion Technologies GmbH, Gabrielenstr. 9, 80636 Munich, Germany; 2Nanion Technologies Inc. 1 Naylon Place, Livingston, NJ, 07039, USA; 3Inst. für Physiologie I, Univ. Bonn, Life and Brain Center, Bonn, Germany; 4Axxam S.p.A., Openzone - via Meucci 3, 20091 Bresso - Milan, Italy; 5OPTEL Consortium funded by EuroTransBio