And elucidating

30 Jan

Specific Aim 2: Determine the extent of disease by performing molecular and functional analyses of hi PSC-CMs.

In the absence of inhibition, NFAT-activated mediators of hypertrophy such as GATA4 and MEF2C were found to be significantly upregulated in HCM i PSC-CMs beginning day 40 post-induction of cardiac differentiation, but not prior to this point.

Taken together, these results indicate that calcineurin-NFAT signaling plays a central role in the development of the HCM phenotype as caused by the Arg663His mutation.

We've differentiated all stablished i PSC lines from all subjects into cardiomyocyte using a modified protocol from that published by Palacek in PNAS 2011.

This protocol increased the yield of cardiomyocytes significantly to consistently greater than 70% beating cardiomyocytes.

We also found increase in torsade formation when the i PSC-CMs are treated with h ERG blockers that are also known to cause increases in arrhythmia in HCM patients.We believe the use of hi PSC-CM from healthy individuals and patients with genetic heart disease can help predict the potential arrhythmic risk in existing or new drug agents that are undergoing FDA evaluation.In this proposal, we will generate and characterize human induced pluripotent stem cell-derived cardiomyocytes (i PSC-CMs) from patients with HCM.Over the past year, we have characterized the pathological phenotypes from i PSCs derived from a 10-patient family cohort with the MYH7 mutation.For example, we found that administration of catecholamine drug norepinephrine causes the formation of torsade de point which is a lethan arrhythmia.This recapitulates the phenotype in patients with HCM receiving catecholamine drugs.Specific Aim 3: Rescue the molecular and functional phenotypes using zinc finger nuclease (ZFN) technology.Over the past year, we have now derived i PSCs from a 10-patient family cohort with the MYH7 mutation.