SBIR-STTR Award

Develop predictive human cardiomyocyte-based all optical assay for cardiotoxicity. Cardiotoxicity is the leading cause of safety-driven withdrawal a all stages and limitation of drug compounds. The current cardiac safety paradigm nonclinical guidance ICH S7B focuses on in vitro hERG assays, which prove to be suboptimal in predicting a compound's cardiotoxicity in human. In the context of high cost and low productivity for new drug discovery, better cardiac safety evaluation becomes a significant public health issue. CSRC, HESI, and FDA are developing a new cardio safety paradigm: Comprehensive In vitro Proarrhythmia Assay (CiPA), which comprises characterizing electrophysiological effects of compounds in iPSC-derived human cardiomyocytes. Q-State's platform can probe electrophysiology of such cells with higher throughput, lower cost, and higher information content (multi-modalities: voltage, Ca2+, pH, and ATP) than current patch clamp assays, and with better temporal and spatial resolution and lower phototoxicity than other optical screening tools. Supported by a $680K NINDS SBIR Phase I award and angel funding, Q-State Biosciences is developing a proprietary turnkey instrument and software system (Optopatch) for simultaneous optical perturbation and optical measurement of membrane voltage in neurons. We propose to adapt Optopach for cardiomyocyte measurements and to develop and validate an Optopatch assay for drug-induced cardiotoxicity in human iPSC- derived cardiomyocytes. Aim 1. Molecular tools. Select the most effective actuators (converting blue light into electrical stimuli for pacing) and reporters (converting AP waveforms into near infrared fluorescence signals) by comparing promoters, trafficking motifs and gene delivery methods, and to thoroughly characterize the sensitivity, speed, photostability, and repeatability of the molecular tools. Aim 2. Instrumentation. Adapt Optopatch hardware and software to optically stimulate and record from > 1, 000 cardiomyocytes simultaneously. Develop fluidics and environmental controls; modify software to support optical pacing, image segmentation, and calculation of cardiac-relevant AP parameters. Aim 3. Assay development and testing. Optimize assays for acute and chronic cardiotoxicity. Validate the assay by quantifying AP changes under a panel of drugs with known mechanisms and cardiac safety profiles. We will provide non-GLP cardiotoxicity screening services to drug discovery companies at the lead identification and optimization stages, supplementing, with the potential to eventually replace, hERG assays. We will also make Optopatch-cardio instrumentation and reagents available to the academic community; to enable mechanistic studies and broaden the applications of electrophysiology platform.

Public Health Relevance Statement:


Public Health Relevance:
Q-State's technology will allow better prediction of a drug candidate's potential adverse effect to human heart than the assays mandated by the current cardiac safety guidance ICH S7B. It will contribute to bringing effective healthcare solutions to market at lower cost and higher productivity, by preventing drugs with cardiotoxicity from reaching the market and preventing the development of valuable therapeutics from being wrongly terminated, which is one of the concerns about the current S7B assays. Making Q-State instrument and reagents available to the academic research community will facilitate better mechanistic studies and therapeutics development to benefit patients with cardiac conditions.

Project Terms:
Action Potentials; Adoption; Award; Biologic Assays; Bioassay; Assay; Biological Assay; Cells; Communities; Drug Preparation; Drug Compounding; drug/agent; Pharmaceutic Preparations; Medication; Drugs; Pharmaceutical Preparations; Neurophysiology / Electrophysiology; Electrophysiology; Electrophysiology (science); Environmental Health Science; Environmental Health; Fluorescence; Genes; Heart; Modern Man; Man (Taxonomy); Human; In Vitro; Institutes; instrumentation; heavy metal lead; heavy metal Pb; Pb element; Lead; Photoradiation; Light; Manuals; Marketing; Methods; neuronal; Neurocyte; Neural Cell; Nerve Unit; Nerve Cells; Neurons; optical; Optics; Patients; Productivity; Public Health; public health medicine (field); Reagent; Research; Risk; Safety; biological signal transduction; Signaling; Signal Transduction Systems; Intracellular Communication and Signaling; Cell Signaling; Cell Communication and Signaling; Signal Transduction; computer program/software; Software; Computer software; Solutions; Technology; Testing; Time; Torsades de Pointes; USFDA; Food and Drug Administration; United States Food and Drug Administration; Iproveratril; Benzeneacetonitrile, alpha-(3-((2-(3, 4-dimethoxyphenyl)ethyl)methylamino)propyl)-3, 4-dimethoxy-alpha-(1-methylethyl)-; Verapamil; Measures; Drug Delivery Systems; Drug Delivery; health care; Healthcare; Promotor; Promoters (Genetics); Promoter; Promotor (Genetics); base; Acute; Chronic; Phase; Variation; Variant; Evaluation; cardiomyocyte; Heart myocyte; Heart Muscle Cells; Cardiocyte; Cardiac Muscle Cells; Cardiac Myocytes; Stimulus; Withdrawal; Measurement; Funding; Therapeutic; Staging; Reporter; tool; instrument; Msec; millisecond; Protocol; Protocols documentation; LOINC Axis 4 System; System; Services; innovative technologies; experience; membrane structure; Membrane; success; V (voltage); voltage; trafficking; Speed; Speed (motion); Modality; assay development; Cardiac Toxicity; Cardiotoxicity; treatment adverse effect; therapy adverse effect; side effect; Treatment Side Effects; Adverse effects; drug discovery; patch clamp; preventing; prevent; Address; Resolution; Small Business Innovation Research; SBIRS (R43/44); SBIR; Small Business Innovation Research Grant; Molecular; Cardiac; Gene Delivery; developmental; Development; Phototoxicity; cell imaging; cellular imaging; NINDS; National Institute of Neurological Disorders and Stroke; cost; software systems; imaging Segmentation; iPSC; iPS; induced pluripotent stem cell; therapeutic development; public health relevance; product development; drug candidate; Regimen; screening

Toxicity accounts for approximately 20% of drug attrition, of which nearly one third is attributed to cardiovascular issues, in particular arrhythmias.1 The cost of bringing a new drug to market can exceed $1.2 billion and require more than 10 years of research. Thus, it is critical to identify cardiotoxicity early in developmen. Current in vitro screening assays for pro-­" arrhythmic drug effects focus on measuring inhibition of the hERG potassium channel, which has been linked to potentially lethal Torsades de Pointes arrhythmias (TdP).2 The hERG assay, however, lacks high sensitivity and specificity: not all QT prolongation is due to block of hERG alone and not all hERG blockers result in QT prolongation or induce TdP. To address these limitations, regulatory bodies have created the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative2, which proposes use of cardiomyocytes derived from human stem cells as an in vitro model for arrhythmogenic drug potential. Establishment of this model for toxicity screening requires detailed characterization of the cardiomyocyte action potential (AP) and calcium transient (CT), the response of APs/CTs to drugs, and the correlation to clinical outcomes in humans. High-­"throughput tools needed for performing these measurements, however, have been lacking. Here we propose to develop a high-­"throughput, all-­"optical electrophysiology platform for cardiotoxicity screening in sem cell-­"derived cardiomyocytes. With our Phase I award, we demonstrated that the Optopatch platform could be used to detect changes in the electrophysiological characteristics of human derived CMs following both acute and chronic drug treatment, albeit with the throughput of a single well per recording. To highly parallelize these measurements, we propose building a 96-­"well plate Optopatch instrument for simultaneous recording of voltage and calcium waveforms under paced conditions from 24 wells. This geometry will provide nearly two orders of magnitude improvement in throughput of our assay. Optopatch constructs will be optimized to allow for incorporation of the actuator and reporter proteins in each cell. We will utilize this platform to screen 50 compounds with known risk scores for Torsades de Pointes in different sources of human cardiomyocytes and use this data to develop a predictive algorithm of arrhythmogenicity.

Public Health Relevance Statement:


Public Health Relevance:
Q­"State's technology will allow better prediction of a drug candidate's potential adverse effects to the human heart than the assays mandated by the current cardiac safety guidance ICH S7B. It will contribute to bringing effective healthcare solutions to market at lower cost and higher productivity, by preventing drugs with cardiotoxicity from reaching the market and preventing the development of valuable therapeutics from being wrongly terminated, which is one of the concerns about the current S7B assays. Making Q-­"State instrumentation and reagents available to the academic research community will facilitate better mechanistic studies and therapeutics development to benefit patients with cardiac conditions.

Project Terms:
Accounting; Action Potentials; adulthood; Adult Human; 21+ years old; Adult; Algorithms; Heart Arrhythmias; Cardiac Arrhythmia; Arrhythmia; Award; Biologic Assays; Bioassay; Assay; Biological Assay; Life Sciences; Bioscience; Biologic Sciences; Biological Sciences; Factor IV; Coagulation Factor IV; Ca++ element; Blood Coagulation Factor IV; Calcium; circulatory system; Heart Vascular; Cardiovascular Organ System; Cardiovascular Body System; Cardiovascular; Cardio-vascular; Cardiovascular system; Cell Body; Cells; Communities; drug treatment; Drug Therapy; Pharmacotherapy; drug/agent; Pharmaceutic Preparations; Medication; Drugs; Pharmaceutical Preparations; electrophysiological; Neurophysiology / Electrophysiology; Electrophysiology; Electrophysiology (science); Engineering; Environmental Health Science; Environmental Health; Heart; Modern Man; Human; In Vitro; Institutes; instrumentation; Membrane Channels; Ionic Channels; Ion Channel; heavy metal lead; heavy metal Pb; Pb element; Lead; Manuals; Marketing; optical; Optics; Paper; Patients; Potassium Ion Channels; K channel; Potassium Channel; Production; Productivity; Proteins; Reagent; Research; Risk; Safety; Sensitivity and Specificity; Computer Software Engineering; Computer Software Development; Software Engineering; Technology; Testing; Time; Torsades de Pointes; Transcription; RNA Expression; Gene Transcription; Genetic Transcription; United States Food and Drug Administration; USFDA; Food and Drug Administration; Vendor; Work; Measures; Healthcare; health care; Blinded; Acute; Chronic; Clinical; Phase; Link; cardiomyocyte; Heart myocyte; Heart Muscle Cells; Cardiocyte; Cardiac Muscle Cells; Cardiac Myocytes; Measurement; Funding; Collaborations; Therapeutic; Staging; Contracting Opportunities; Contracts; Reporter; tool; instrument; support vector machine; statistical learning; kernel methods; Machine Learning; Hour; Frequencies; Protocol; Protocols documentation; Source; Techniques; System; Services; success; voltage; Toxicities; Toxic effect; Reporting; Position; Positioning Attribute; Modeling; Human Nature; Human Characteristics; response; Cardiotoxic; Cardiac Toxicity; Cardiotoxicity; treatment adverse effect; therapy adverse effect; side effect; Treatment Side Effects; Adverse effects; drug discovery; patch clamp; Provider; preventing; prevent; Address; Data; Detection; molecular signature; molecular profile; Molecular Fingerprinting; Expression Profiling; Molecular Profiling; Resolution; in vitro Model; Cell Maturation; Characteristics; Molecular; Cardiac; developmental; Development; Phototoxicity; imaging; Image; human stem cells; cost; Outcome; clinically relevant; clinical relevance; novel therapeutics; novel therapy; novel drugs; novel drug treatments; next generation therapeutics; new therapy; new therapeutics; new drugs; new drug treatments; induced pluripotent stem cell; iPSCs; iPSC; iPS; spatiotemporal; therapeutic development; public health relevance; stem cell biology; drug candidate; phase 1 study; Phase I Study; optogenetics; screening; Geometry; rate of change; prediction algorithm; predictor algorithm; predictive algorithm