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Research Interests. The Role of Catecholamines in Cardiac Lineage Commitment Electrophysiological and pharmacological characterization of ES and iPS cell-derived cardiomyocytes. Martin Lehmann , PhD . Institute for Neurophysiology University of Cologne lehmannm@uni-koeln.de.
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Research Interests The Roleof Catecholamines in Cardiac Lineage Commitment Electrophysiological and pharmacological characterization of ES and iPS cell-derived cardiomyocytes Martin Lehmann, PhD. Institute forNeurophysiology University of Cologne lehmannm@uni-koeln.de
I. The Roleof Catecholamines in Cardiac Lineage Commitment • Catecholamines play an essential role in heartfunctionofthe adult heart • SeveredevelopmentaleffectsofCatecholamine Synthesis Gene knockout (TH and DBH) (Kobayashi et al., 1995; Zhou et al., 1995, Thomas et al., 1995) • Paracrineactionof catecholamines at developmentalstagebefore neuronal innervation was suggested • In Contrast: In vivo Pnmt k.o. (Ebert et al., 2004) did not leadto fetal mortality • Noradrenalineismostimportantamongstthe catecholamines in respecttocardiacdevelopment Research Interests, Martin Lehmann, PhD. lehmannm@uni-koeln.de
I. The Roleof Catecholamines in Cardiac Lineage Commitment IntracellularCatecholamineSynthesis * Reserpine- Mechanismofaction * * http://what-when-how.com/wp-content/uploads/2012/04/tmp14754_thumb1.jpg Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
I. The Roleof Catecholamines in Cardiac Lineage Commitment • Reserpine-inducedcatecholaminedepletion, reduced cardiomyocyte differentiationefficiency d2 d4 d10 d14+Puro Control DMSO Reserpine Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de d2+d4: 50 µm; d10+d14: 200 µm
I. The Roleof Catecholamines in Cardiac Lineage Commitment • ReserpineReduces Numbers ofBeating EBs Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
I. The Roleof Catecholamines in Cardiac Lineage Commitment Cardiac TroponinT aActinin Scalebars: 50 µm • Expression ofcardiacmarkersissignificantlyreduced after catecholaminedepletion qPCRsby Dr. V. Wagh Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
I. The Roleof Catecholamines in Cardiac Lineage Commitment • Interestingly, catecholaminedepletionpromotes neuronal lineagecommitment Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
I. The Roleof Catecholamines in Cardiac Lineage Commitment • Catecholamines play a crucialrole in cardiacdifferentiationexhibitingtheiraction in a paracrinefashion • The criticalperiodforcatecholamineactionisbeforeday 6 • a- andb-adrenergicsignalingiscriticalduringcardiacdifferetiation. Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
II. Electrophysiological and pharmacological characterization of ES and iPS cell-derived cardiomyocytes
MEA-basedhuman ES or iPS cell-derivedCardiomyocyte (rESCM) screen Project Aim - Drugs canhaveunexpectedcardio-activesideeffects (e.g. torsade de pointestachycardia, TdP) Pharmacological Screening Tool todetectpossiblecardio-activeeffectsbeforecost-intensive clinicalphase WhyMultielectrodeArrays (MEA)? • non-invasive extracellularmeasurement (offieldpotentials (FPs)) • long-term recordings • allowsforrecordingof 2D excitationandconduction (slices, monolayer) Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
Multielectrode Array (MEA) Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
Multielectrode Array (MEA) Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
LabView-based Analysis Tool MEA QT-Tool Data Display Data Processing Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
MEA QT-Tool Analysis Results Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
FP/f Correlations-PhysiologicPositive Chronotropic Conditions Normalizedaveragedvalues • Negative (linear) FP/f correlation upon physiologic positive chronotropic stimulation • Normalizedandaveragedvaluepairsarerepresentativefor a populationof EBs Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
FP/f Correlations-PhysiologicNegative Chronotropic Conditions FP/f correlationduring negative chronotropic stimulation Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
FP/f Correlations-QT-prolongingconditions Hypothetically, QT-time andrepolarizationprolongingdrugsshouldchangethe FP/f correlationtowardless negative values, e.g. E4031 Antiarryhthmicclass III drug(K+/HERG-Channel blocker) positive FP/f correlation Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
II. Electrophysiological and pharmacological characterization of ES and iPS cell-derived cardiomyocytes Conclusions: • Drug effects on different EBs/cellsiscomparable (normalization!!) • EB-to-EB comparisonhastobetakenwith care (e.g. Long-QT-iPS) • A pharmacologicalscreenisfeasiblewith MEA irrespectiveofcell type (ES oriPS) Research Interests, Martin Lehmann, PhD.; lehmannm@uni-koeln.de
