FUJIFILM Cellular Dynamics, Inc. and Merck Research Laboratories Sponsored Presentation

T.K. Feaster, PhDHaoyu Zeng, PhD

March 11, 2019

Session OverviewPart I

• Dr. T.K. Feaster • i) hiPSC-CMs in Safety and Toxicity Testing: Genetic Diversity• ii) hiPSC-CMs in Safety and Toxicity Testing: Disease Modeling

Part II• Dr. Haoyu Zeng

• Resolving the Reversed Rate Effect of Calcium Channel Blockers on hiPSC-CMs and the Impact on In Vitro Cardiac Safety Evaluation

Outline of Presentation

1. FUJIFILM Cellular Dynamics, Inc. and iPSC Technology

2. Genetic Diversity Panel

3. Disease Models• CPVT - Catecholaminergic Polymorphic Ventricular Tachycardia• BrS - Brugada Syndrome Type 3• DCM - Laminin-Related Dilated Cardiomyopathy • HCM - Hypertrophic Cardiomyopathy

4. Summary

FUJIFILM Cellular Dynamics iPSC Technology

Applications Support

BasicScience

Healthy donor orPatient-derived

DrugDiscovery

CellTherapy

BRAIN & NERVOUS SYSTEMiCell Astrocytes

iCell DopaNeurons

iCell GABANeurons

iCell GlutaNeurons

iCell Motor Neurons

iCell Microglia

HEART & VASCULARiCell Cardiomyocytes

iCell Endothelial Cells

iCell Cardiac Progenitor Cells

LIVERiCell Hepatocytes 2.0

iCell Hepatoblasts

OCULARiCell Retinal Pigment Epithelial Cells

iCell Hematopoietic Progenitor Cells

iCell Macrophages

BLOOD & IMMUNE

iCell® Products: Unprecedented Access to Human Cells

iCell CardiomyocytesExcitation-Contraction Coupling

Electrical Activity Calcium Handling Mechanical Beating

iCell Cardiomyocytes are spontaneously beating cells that are ideal for interrogation of excitation-contraction coupling

iCell Cardiomyocytes Genetic Diversity Panel

Data Generation

Understand compound effectsacross genetic backgrounds

iCell Cardiomyocytes Genetic Diversity Panel to explore incidence and frequency of compound response across genetic backgrounds

As published in Grimm et al. 2018 ALTEX, Huo et al. 2018 ToxSci andBlanchette et al 2018 Clin Pharmacol Ther

“Clinical Trial‐in‐a‐Dish” in vitro CohortiCell Cardiomyocytes available from

30 healthy donors

SelectCohort

https://www.ncbi.nlm.nih.gov/pubmed/29999168

iCell Cardiomyocytes Genetic Diversity Panel

• Investigate compound effects (efficacy and safety) across a cohort of healthy donors• Gain insight into incidence and frequency of drug response at clinically relevant

concentrations• Select the appropriate cohort to fit your experimental and population needs

iCell Cardiomyocytes Genetic Diversity Panel Demographic

https://www.ncbi.nlm.nih.gov/pubmed/29999168

iCell Cardiomyocytes genetic diversity panel enables researchers to evaluate compound response across multiple donors.

Pharmacological Response Across Different Backgrounds

6 Donor Panel of iCell Cardiomyocytes27 Donor Panel of iCell Cardiomyocytes

http://asecho.org/wordpress/wp-content/uploads/2016/10/A.-Klein-Cardiotoxic-Effects-of-Chemotherapeutic-Drugs.pdf

Doxorubicin [10M]

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Doxorubicin [0.1M]

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15LegendLegendLegendLegendLegendLegend

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Doxorubicin [1M]

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Dox Dox Dox

Donor 1 Cardiomyocytes = Green

Donor 2 Cardiomyocytes = Brown

Donor 3 Cardiomyocytes = Red

Donor 4 Cardiomyocytes = Blue

Donor 5 Cardiomyocytes = Purple

Donor 6 Cardiomyocytes = Grey

DMSO Control

Drug

Doxorubicin-induced Cardiomyocyte Death in a Time and Concentration Dependent Manner

Negligible effect with acute treatment (1 hr)

1 hr Post-TreatmentDonor 1

Donor 2

Donor 3

Donor 4

Donor 5

Donor 6

DMSO

10 µM

1 µM

0.1 µM

DMSO

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1 µM

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DMSO

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Marked effect with chronic treatment (24 hr)

24 hr Post-Treatment

DMSO

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DMSO

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1 µM

0.1 µM

DMSO

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1 µM

0.1 µM

Donor 1

Donor 2

Donor 3

Donor 4

Donor 5

Donor 6

Marked effect with chronic treatment (72 hr)

72 hr Post-Treatment

DMSO

10 µM

1 µM

0.1 µM

DMSO

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DMSO

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Donor 1

Donor 2

Donor 3

Donor 4

Donor 5

Donor 6

Doxorubicin Alters Cardiomyocyte Function and Lead to Cell Death at Higher Concentrations and Durations

Sunitinib [0.1M]

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Sunitinib [10M]

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Sunit Sunit Sunit

Sunitinib has no Effect on Cardiomyocyte Viability

Donor 1 Cardiomyocytes = Green

Donor 2 Cardiomyocytes = Brown

Donor 3 Cardiomyocytes = Red

Donor 4 Cardiomyocytes = Blue

Donor 5 Cardiomyocytes = Purple

Donor 6 Cardiomyocytes = Grey

DMSO Control

Drug

Observable effects with acute exposure

DMSO

10 µM

1 µM

0.1 µM

DMSO

10 µM

1 µM

0.1 µM

DMSO

10 µM

1 µM

0.1 µM

Donor 1

Donor 2

Donor 3

Donor 4

Donor 5

Donor 6

More pronounced with chronic treatment

DMSO

10 µM

1 µM

0.1 µM

DMSO

10 µM

1 µM

0.1 µM

DMSO

10 µM

1 µM

0.1 µM

Donor 1

Donor 2

Donor 3

Donor 4

Donor 5

Donor 6

More pronounced with chronic treatment

DMSO

10 µM

1 µM

0.1 µM

DMSO

10 µM

1 µM

0.1 µM

DMSO

10 µM

1 µM

0.1 µM

Donor 1

Donor 2

Donor 3

Donor 4

Donor 5

Donor 6

1 hr Post-Treatment 24 hr Post-Treatment 72 hr Post-Treatment

Sunitinib Alters Cardiomyocyte Function without Leading to Cell Death

i) Summary

•iCell Cardiomyocytes Genetic Diversity Panel enables assessment of multiple compounds and concentrations across diverse donors.

•iCell Cardiomyocytes display cancer therapeutic-related cardiac dysfunction to doxorubicin (Type I) and sunitinib (Type II).

INNATE ENGINEERED INDUCED

Reprogrammingto iPSC

iPSCDifferentiation

Healthy Donor Donor with Genetic Disease

Controls

Healthy Conditions

Disease-inducingConditionsGenome

Engineering

Phenotypic andFunctional Analysis

Healthy Donor Healthy Donor

Disease Modeling with iPSC derived Cells – “Disease-in-a-Dish”

Donor with Genetic Disease

iCell Cardiomyocytes CPVT (E2311D) display abnormal cardiac function.

• CPVT (E2311D) Cardiomyocytes display abnormal electrophysiological properties.

• CPVT (E2311D) Cardiomyocytes display reduced beat rate and increased beat rate irregularity.

Disease Model: Isogenic Pair of Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT RYR2 E2311D) Cardiomyocytes and Corrected Cardiomyocytes

iCell Cardiomyocytes BrS3 (G490R) display abnormal function and sensitivity to calcium channel modulators.

• BrS3 (G490R) Cardiomyocytes display abnormal baseline properties.

• Reduced calcium channel function results in increased sensitivity to calcium blockers in BrS3 (G490R) Cardiomyocytes.

Disease Model: Isogenic Pair of Brugada Syndrome (BrS3 CACNA1C G490R) Cardiomyocytes and Corrected Cardiomyocytes

• DCM (LMNA L35P) Cardiomyocytes display comparable morphology.

• DCM (LMNA L35P) Cardiomyocytes display reduced contraction (impedance).

• DCM (LMNA L35P) Cardiomyocytes display comparable calcium handling at baseline.

iCell Cardiomyocytes DCM (L35P) display reduced contractile properties.

Disease Model: Isogenic Pair of Dilated Cardiomyopathy (DCM LMNAL35P) Cardiomyocytes and Corrected Cardiomyocytes

• HCM (R403Q) Cardiomyocytes display abnormal baseline properties.

• HCM (R403Q) Cardiomyocytes display increased contraction (impedance) and arrhythmic events.

• HCM (R403Q) Cardiomyocytes display alerted calcium handling including prolong calcium kinetics.

Disease Model: Isogenic Pair of Hypertrophic Cardiomyopathy (HCM MYH7 R403Q) Cardiomyocytes and Corrected Cardiomyocytes

iCell Cardiomyocytes HCM (R403Q) display HCM phenotype including abnormal electrophysiological, calcium handling and contractile properties.

ii) Summary

•iPSC-derived disease models recapitulate classical disease hallmarks in vitro.

•Utilization of an isogenic control enables investigation of the direct consequences of the mutation.

•iPSC-derived disease models respond appropriately to specific pharmacological challenge.

Thank you