Visualization of SERCA2a-PLB Interaction: Mechanisms for Enhanced Cardiac Contractility
Heart Failure with Reduced Ejection Fraction (HFrEF) is a global health concern affecting millions worldwide, characterized by inefficient blood pumping due to left ventricular enlargement. The Meyer lab's innovative approach utilizes a circularly permutated green fluorescent protein (cpGFP) to investigate the SERCA2a-PLB interaction in various cell types. The novel SERCA2a-cpGFP-PLB construct, expressed in HEK 293 cells, H9 cells, and induced pluripotent stem cells, enables selective interference studies with fluorescence as a functional indicator. Using a Nikon A1R FLIM Confocal Microscope, the study aimed to visualize the temporal interaction between SERCA2a and PLB during contraction and relaxation in diverse cell types. Manipulating cytosolic calcium levels through forskolin, isoproterenol, and stimulation rate variations will elucidate the impact on contractility. The hypothesis proposes that Ca2+-dependent PLB deinhibition of SERCA2a, mimicking effects of forskolin and isoproterenol, can enhance contractility, correlating higher heart rates with stronger contractions. This research addresses a critical gap in understanding HFrEF cellular mechanisms. Insights into SERCA2a-PLB dynamics may guide the development of targeted therapies, especially considering alterations in these proteins in HFrEF patients. The potential benefits of increased heart rates, supported by recent clinical evidence, suggest novel treatments for HFrEF. Dissemination through scientific posters and papers aims to contribute valuable insights to the scientific community, fostering a better understanding of cardiac function and potential pharmacological interventions for heart failure.