In ventricular cardiomyocytes, transverse-tubules (T-tubules) concentrate L-type calcium channels (LTCC) which approximate and form dyads with ryanodine receptors (RyR2) at sarcoplasmic reticulum membrane. It is not well understood how the dyads form. The overall objective of this application is to identify the key components of dyad organization in healthy and failing hearts. My central hypothesis is that the membrane deformation protein BIN1 creates microdomains within T- tubule microfolds, organizing LTCC clusters and recruiting RyR2 receptors for proper dyad formation, facilitating synchronized calcium-induced-calcium-release and limiting arrhythmias. Furthermore, BIN1 is known to decrease in acquired heart failure, and heart failure can result when these BIN1-orgnized microdomains are disrupted. The rationale that underlies the proposed research is that BIN1-based impaired regulation of dyad structure and function is a reversible aspect of heart failure progression. The first of three aims is to determine, in physiologically normal cardiomyocytes, whether and how cardiac BIN1 is responsible for organizing LTCC-RyR2 dyads. Building on preliminary data, in adult mouse (with or without Bin1 deletion) and human cardiomyocytes, we will use super-resolution fluorescent imaging to identify LTCC-RyR2 localization at BIN1-induced microdomains, and test the role of actin cytoskeleton in LTCC-RyR2 complex formation and function.
The second aim i s to determine how BIN1 dependent microdomains are disrupted in heart failure. Building on preliminary data, we will use human and mouse models of heart failure to quantify BIN1 splicing and isoform expression, T- tubule folds, and LTCC-RyR2 complex formation and function, as well as the rescue ability of exogenous BIN1. We will also study whether the mechanism of reduced Bin1 transcription and aberrant splicing in heart failure is due to ?-adrenergic sympathetic dysregulation.
The third aim i s to determine whether ?-adrenergic blockade can rescue BIN1-microdomains and limit arrhythmias and heart failure development. Building on preliminary data, we will investigate whether Bin1 deleted mice develop arrhythmia and heart failure when stressed, which can be rescued by ?-adrenergic blockade. Our contribution here is expected to be a detailed understanding of how BIN1-induced LTCC- RyR2 microdomains are organized and regulated in both normal and diseased hearts. This contribution is significant because it will identify a new approach to ameliorate heart failure progression by preserving the BIN1 microdomains that are critical to normal heart function. The proposed research is innovative, in our opinion, because it introduces the relatively unknown BIN1 as a determinant of T- tubule microdomains thus helping regulate cardiac dyads and calcium transients.
The proposed research is relevant to public health and NIH?s mission because it will introduce new targets for therapeutic interventions capable of limiting the progression of heart failure in millions of Americans. Heart failure is the major cardiac syndrome in the United States and a better molecular understanding of how cardiomyocyes in failing hearts progressively lose their function is a critical step to developing new medical solutions.
|Basheer, Wassim A; Fu, Ying; Shimura, Daisuke et al. (2018) Stress response protein GJA1-20k promotes mitochondrial biogenesis, metabolic quiescence, and cardioprotection against ischemia/reperfusion injury. JCI Insight 3:|
|Hong, TingTing; Shaw, Robin M (2017) Cardiac T-Tubule Microanatomy and Function. Physiol Rev 97:227-252|
|Xu, Bing; Fu, Ying; Liu, Yan et al. (2017) The ESCRT-III pathway facilitates cardiomyocyte release of cBIN1-containing microparticles. PLoS Biol 15:e2002354|
|Fu, Ying; Zhang, Shan-Shan; Xiao, Shaohua et al. (2017) Cx43 Isoform GJA1-20k Promotes Microtubule Dependent Mitochondrial Transport. Front Physiol 8:905|
|Basheer, Wassim A; Xiao, Shaohua; Epifantseva, Irina et al. (2017) GJA1-20k Arranges Actin to Guide Cx43 Delivery to Cardiac Intercalated Discs. Circ Res 121:1069-1080|
|Zhou, Kang; Hong, Tingting (2016) Cardiac BIN1 (cBIN1) is a regulator of cardiac contractile function and an emerging biomarker of heart muscle health. Sci China Life Sci :|
|Fu, Ying; Shaw, Seiji A; Naami, Robert et al. (2016) Isoproterenol Promotes Rapid Ryanodine Receptor Movement to Bridging Integrator 1 (BIN1)-Organized Dyads. Circulation 133:388-97|
|Fu, Ying; Hong, TingTing (2016) BIN1 regulates dynamic t-tubule membrane. Biochim Biophys Acta 1863:1839-47|