Hipsc-Cardiovascular Microtissue Platform Integrated with Magnetoresistance Sensors for the Study of Static Mechanics, and Mechanical Adaptation Related to the Onset and Progression of Inherited Cardiovascular Disease( I )

Cardiovascular diseases (CVDs) accounted for 31% of all deaths globally1. Both arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) and vascular Ehlers-Danlos Syndrome (vEDS), are inherited disease affected by mechanical stress and cause early death. Using human pluripotent stem cell (hiPSC) derived cardiovascular cells to study ARVD/C and vEDS can faithfully recapitulate key functional properties of cardiac and cardiovascular system in vivo. A sensing system integrated with a microtissue platform that can give a real-time readout of the kinetic force generation will enable significant advances in the understanding of tissue mechanics and related disease processes. Therefore, this proposal will develop new technologies in the area of magnetic microfabricated tissue gauges (µTUGs) sensor systems, and hiPSC derived cardiomyocytes (CMs)/vascular smooth muscle cells (vSMCs) in collaboration with the Johns Hopkins University (JHU). In first year, we will generate spin-valve multilayer stripe GMR elements and characterizing their magnetoresistance for an optimal design sensitive to detect microtissue contraction. By adjusting the bridge configuration and circuit design will increase S/N ratio for sensitive response. We will test the biocompatibility, reliability of the sensor in long-term culture condition. We will make the ARVD CM tissues (CMTs) with CM desmosome mutation (PKP2, DSG2, and SCN5A) and vEDS vSMC tissues (SMTs) with COL3A1 gene mutation. We will analyze structural, gene expression, proteomics and metabolic studies under basal condition. In the second year, we will develop software for signal processing and acquisition of multiple GMR signals by multichannel lock-in Amplifier for high throughput data analysis. CRISPR-corrected cell lines will be available from JHU and we will also make CMTs/SMTs, assess gene/phenotype information and compared isogenic control with individual disease counterpart. The spontaneous or electrical-paced contraction kinetics output of individual ARVD CMTs will then be assessed by µTUG -GMR sensor arrays and compare the different desmosome gene mutation with the isogenic CMTs controls. vEDS SMT will be under different basal loading and their developed force as compared with their isogenic control will be studies. In the third year, we will use with uncoupler reagent to separate the cell contribution to tissue mechanical property and analyze the influence of abnormal extracellular matrix remolding of vEDS to disease pathogenesis. We will do linear mechanical stretching to get the stress versus strain readout before/after uncoupler treatment and then compare the readout of vEDS SMT with isogenic control. To ensure the ARVD/C -CM with the disease phenotype, we will use do biochemical treatment such as 5 F media to coactivate normal (PPAR-α) and Pathogenic (PPAR-γ) pathway and characterize the dynamic force response of ARVD/C CMTs on µTUG GMR sensor and compare their gene profile. In the fourth year, we will do acute and chronic mechanical stretching to vSMTs and CMTs. Force generation, structural/mechanical integrity, extracellular remolding, molecular and metabolic will be studied to see tissue adaptation in response to different frequency, magnitude, intensity of mechanical stress. The µTUG GMR sensor platform can give a real-time readout of the kinetic force generation will enable significant advances in the understanding of tissue mechanics and related disease processes.