Methods for improving relaxation of striated muscle cells - Patents.com

Abstract

The inventors developed conditions that allow efficient detection of differences in the relaxation phase of cardiomyocytes associated with increased stiffness of cardiomyocytes. The inventors used libraries of patient-specific human induced pluripotent stem cells (hiPSCs) from healthy donors or from donors with mutations associated with increased passive stiffness in cardiomyocytes (i.e., MYH7 and BRAF mutations) as well as hypertrophic cardiomyopathy, a condition typically associated with impaired diastolic function. The inventors performed a high-throughput screen on hiPSC-derived cardiac cells to identify microRNAs capable of modifying the relaxation rate of cardiomyocytes. In particular, the inventors set up a large-scale functional genomics using miRNA screening. All identified miRNAs were tested for their effect on cardiac cell movement and calcium transients. The miRNAs with the highest impact were specifically tested for ECT, and changes in diastolic function were measured and compared with the results obtained at the cellular level. The inventors engineered the most interesting "hits" in a 3D model using a readout similar to the primary assay. We test the impact of the positive "hits" in the mechanical model (developed in the discovery part) and establish the physiological and biochemical mechanism of action of the main proteins identified. We finally identify two promising miRNAs that can also be used to improve striated muscle cell relaxation and, more generally, to alleviate symptoms related to striated muscle stiffness, especially in the context of heart failure with preserved ejection fraction (HFpEF).

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