THE CardioECR/iCELL SYSTEM @ CHARLES RIVER A tool for - - PowerPoint PPT Presentation

THE CardioECR/iCELL SYSTEM @ CHARLES RIVER

A tool for excitation-contraction coupling in cardiac safety and drug discovery

EVERY STEP OF THE WAY EVERY STEP OF THE WAY

MECHANISTIC ANALYSIS OF CONTRACTION

INOTROPIC EFFECTS

TOOLS

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xCELLigence RTCA CardioECR iCELL cardiomyocytes Drugs: Blebbistatin Nifedipine FPL 64176 Omecamtiv mecarbil

THE CardioECR/ICell SYSTEM

Cell Index = (Impedance @ time t – Impedance @ time 0 ) / 15 Ω

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N=45 Cell Index = (Rtn-Rt0)/15Ω

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THE IMPEDANCE SIGNAL

MATLAB Analysis Raw data Local averages Grand Average

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MOTION-DEPENDENT EVENTS: BLEBBISTATIN

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EXCITATION-CONTRACTION COUPLING: L-TYPE CA2+ MODULATORS

NIFEDIPINE (BLOCKER) FPL 64176 (AGONIST)

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MOTION-DEPENDENT EVENTS: OMECAMTIV MECARBIL

Contraction: Positive inotropy Impedance recordings (CardioECR) 50,000 iCELL2 cardiomyocytes CL: 1500 ms Omecamtiv mecarbil 1 µM

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CONCLUSION

Discovery Application

• Enhanced software algorithms allow detailed investigation of excitation-

contraction coupling and contraction mechanisms.

• The system responds to known inotropic agents in a predictive way and

could be a valuable tool to investigate contraction in a medium throughput plate-based assay.

MECHANISTIC ANALYSIS OF ARRYTHMIC LIABILITY

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ARRHYTHMIC LIABILITY

You want this … FIELD POTENTIAL IMPEDANCE

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ARRHYTHMIC LIABILITY

Ibutilide 100 nM

But some times you get this at the Cmax ( 140 nM ) FIELD POTENTIAL IMPEDANCE

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ARRHYTHMIC LIABILITY

Moxifloxacin: Cmax 11 µM Baseline 10 µM 30 µM 100 µM 300 µM

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ARRHYTHMIC LIABILITY

The CardioECR/iCELL system detects delayed repolarization and arrhythmias

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CONCLUSION

• The CardioECR/iCELL system is a valuable tool to investigate proarrhythmic liability (CiPA)
• The system responds to known pharmacological agents in a predictive way.
• When tailored with appropriate disease cell models the CardioECR/iCell system is a valuable tool to generate

cardiac disease biomarkers. Cardiac Safety Application

MECHANISTIC ANALYSIS OF CARDIOTOXICITY

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CARDIOTOXIC DRUGS: LONG TERM EFFECTS

Vehicle: 8 days

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CARDIOTOXIC DRUGS: LONG TERM EFFECTS

Vehicle and Doxorubicin: 8 days

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CARDIOTOXIC DRUGS: LONG TERM EFFECTS

Effect of Tyrosine Kinase Inhibitors on basal impedance

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CARDIOTOXIC DRUGS: LONG TERM EFFECTS

Crizotinib: Effects at Therapeutic Doses

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CARDIOTOXIC DRUGS: LONG TERM EFFECTS

Erlotinib: Supratherapeutic Effects

Ouabain Shortens the FPD (QT-like Signal)

• Cardiac glycoside, increases

intracellular calcium

• Low dose increases contractile

force, has been used as heart failure therapy

• High dose causes cardiac arrest

(used for poison arrows)

Ouabain Does Not Inhibit Ion Channel Currents

0.0 10.0 20.0 30.0 40.0 50.0 60.0 70.0 80.0 90.0 100.0 0.03 0.1 0.3 0.03 0.1 0.3 Percent Inhibition Concentration (µM)

Ion Channel Inhibition

hERG Nav1.5 KvLQT1/minK Kir2.1 Cav1.2 Ouabain Digoxin

Ouabain Toxicity is Revealed in Contraction Measure (Impedance)

1 hr 2 hr 3 hr 4 hr Vehicle 2.5 nM 5 nM 10 nM 20 nM 40 nM

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OVERALL CONCLUSIONS

• The CardioECR/iCell system is a valuable tool to investigate:
• Contractility
• Proarrhythmic liability (CiPA)
• Cardiotoxicity
• Medium throughput analysis of human cells (integrated system) detects

liability that would be missed using in vitro ion screening strategies alone

CONTACT US

Andrew Bruening-Wright, PhD Principal Scientist Safety Assessment | Charles River 251 Ballardvale Street Wilmington, MA 01887 askcharlesriver@crl.com www.criver.com 877.CRIVER.1