LightKick: Novel bioengineering system for activity-dependent acceleration of functional maturation of human stem cell-derived cardiomyocytes

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ABSTRACT Human iPSC-derived cardiomyocytes offer infinite possibilities for cardiovascular drug discovery and regenerative medicine, but only if they reflect the physiology of an adult human heart. Current protocols produce cardiomyocytes with embryonic rather than adult profiles, severely limiting their utility for the adult population. Recent studies suggest that bioengineering approaches aimed at re-creating environmental cues in a dish can lead to marked improvements in cardiomyocytes’ maturation profiles. New technological tools are required to translate these findings into the practice. To enable scientists in academia and biotech/pharmaceutical industry to generate more mature hiPSC-derived cardiomyocytes on demand, we propose to design, engineer, fabricate, and validate a novel bioengineering hardware system that will optically control the pacing of hiPSC-derived cardiomyocytes over extended periods of time to drive their maturation toward the adult phenotype in an activity-dependent manner. The proposed maturation-promoting system will utilize our proprietary graphene-mediated optical stimulation technology that allows fast, reversible, and physiologically relevant activation of cells. Our technology has several critical advantages, as (1) it requires neither genetic modifications (vs. optogenetics) nor wires/electrodes (vs. electric field stimulation), (2) it offers an uncharted flexibility for tunable light- controlled cell stimulation protocols, and (3) it is compliant with high-throughput and scalability requirements. The components of the proposed system (LightKick) will include graphene-coated microplates (G-plates), a multi-LED unit to illuminate cells cultured in G-plates inside a cell incubator, and an external multiparametric programmable controller to define illumination parameters. G-plates have already been developed and validated for optical stimulation and all-optical assays. We will further optimize G-plates in terms of the substrate rigidity and micropatterning. The development of two other components, together comprising an optical stimulation module, will represent the main engineering efforts of the proposed project. To tailor this module for long-term optical stimulation, we will take into consideration recently developed light emitters, optoelectronic properties of G-plates, biophysical properties of cardiomyocytes, and long-term cell culturing conditions. Extensive technical capabilities of our optical stimulation module will offer (1) the scalability (by delivering stimulation signals in few wells or few plates on demand); (2) wireless monitoring of the module and remote control of optical pacing; and (3) the flexible tunability across the wide range of physiologically-relevant light illumination parameters using a user-friendly interactive GUI software. Importantly, long-term optical stimulation of hiPSC-derived cardiomyocytes enabled by the LightKick would be fully compatible with many environmental cues that might play in the maturation process. The LightKick system will allow stem cell experts (1) to develop efficient and individualized cell stimulation patterns for producing adult-like patient-specific hiPSC-derived cells, and (2) to easily combine dynamic electric environmental cues with other environmental cues, further improving multiparametric maturation-promoting protocols. We anticipate that the LightKick system will dramatically simplify and, thus, accelerate the process of finding fundamental and practical answers to the maturation problem.