Ankyrin B (AnkB) is an 'adaptor' protein that anchors several membrane proteins to the cytoskeleton. AnkB deficiency emerged as an important pro-arrhythmic factor a few years ago, when it was found that AnkB loss- of-function mutations generate human long QT syndrome type 4 (LQT4), the only LQT produced by alterations in a protein other than an ion channel. Besides LQT4, humans with AnkB loss-of-function mutations display a complex cardiac phenotype that also includes bradycardia, stress-induced ventricular arrhythmias and sudden cardiac death. The mechanisms responsible for such a phenotype are largely unknown. AnkB protein expression and distribution are drastically altered in the infarct border zone after myocardial infarction (MI). This may result in a mechanism for post-MI remodeling and arrhythmogenesis similar to that found in patients with AnkB loss-of-function mutations. Direct interaction with AnkB is required for the membrane targeting and stability of the Na/Ca exchanger (NCX) and Na/K-ATPase (NKA), which are essential in the regulation of cardiac [Na]i and [Ca]i and thus contractility and potential for triggered arrhythmias. The severity of the human cardiac phenotype generated by various AnkB loss-of-function mutants directly relates to the inability of those mutants to target NCX and NKA correctly to the cardiac myocyte sarcolemma. Thus, altered NCX and NKA expression and membrane distribution are key to the cardiac phenotype generated by AnkB loss-of-function. This phenotype is largely reproduced in mice heterozygous for a null mutation in AnkB (AnkB mice). Myocytes from AnkB mice show modestly reduced NCX and NKA protein expression, predominantly at the T-tubules, larger cellular and sarcoplasmic reticulum (SR) Ca load and increased frequency of early afterdepolarizations (EAD) and delayed afterdepolarizations (DAD). Despite its physiological and pathophysiological significance, the role of AnkB in regulating cardiac [Ca]i and arrhythmogenesis is poorly understood. The overall goal of this proposal is to decipher the mechanisms responsible for altered cardiac Ca regulation and triggered arrhythmias induced by AnkB loss-of-function (inherited and acquired). We will combine measurements of [Na]i and [Ca]i (in the bulk and junctional cleft), patch-clamp and molecular biology techniques in isolated cardiac myocytes in three Specific Aims. First, I will test several specific hypotheses aimed at understanding how AnkB affects intracellular Ca in the heart.
Aim 2 will focus on the mechanisms responsible for the occurrence of early and delayed afterdepolarizations in myocytes with AnkB deficiency. In the final aim I will test the hypothesis that AnkB proteolysis by calpain and the ensuing remodeling in NCX and NKA is a more general mechanism for triggered ventricular arrhythmias. These studies will both advance our understanding of how AnkB affects cardiac [Ca]i regulation and will provide key mechanistic information that could lead to the development of new treatments for patients with ventricular arrhythmias associated with inherited or acquired AnkB loss-of-function.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
7R01HL109501-04
Application #
8787535
Study Section
Special Emphasis Panel (ZRG1-CVRS-F (02))
Program Officer
Krull, Holly
Project Start
2011-09-01
Project End
2016-06-30
Budget Start
2014-03-27
Budget End
2014-06-30
Support Year
4
Fiscal Year
2013
Total Cost
$339,558
Indirect Cost
$111,798
Name
University of Kentucky
Department
Pharmacology
Type
Schools of Medicine
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Stewart, Bradley D; Scott, Caitlin E; McCoy, Thomas P et al. (2018) Computational modeling of amylin-induced calcium dysregulation in rat ventricular cardiomyocytes. Cell Calcium 71:65-74
Popescu, Iuliana; Yin, Guo; Velmurugan, Sathya et al. (2018) Lower sarcoplasmic reticulum Ca2+ threshold for triggering afterdepolarizations in diabetic rat hearts. Heart Rhythm :
Van Steenbergen, Anne; Balteau, Magali; Ginion, Audrey et al. (2017) Sodium-myoinositol cotransporter-1, SMIT1, mediates the production of reactive oxygen species induced by hyperglycemia in the heart. Sci Rep 7:41166
Liu, Miao; Verma, Nirmal; Peng, Xiaoli et al. (2016) Hyperamylinemia Increases IL-1? Synthesis in the Heart via Peroxidative Sarcolemmal Injury. Diabetes 65:2772-83
Popescu, Iuliana; Galice, Samuel; Mohler, Peter J et al. (2016) Elevated local [Ca2+] and CaMKII promote spontaneous Ca2+ release in ankyrin-B-deficient hearts. Cardiovasc Res 111:287-94
Despa, Sanda; Vigmond, Edward (2016) From Single Myocyte to Whole Heart: The Intricate Dance of Electrophysiology and Modeling. Circ Res 118:184-6
Lambert, Rebekah; Srodulski, Sarah; Peng, Xiaoli et al. (2015) Intracellular Na+ Concentration ([Na+]i) Is Elevated in Diabetic Hearts Due to Enhanced Na+-Glucose Cotransport. J Am Heart Assoc 4:e002183
Despa, Sanda; Shui, Bo; Bossuyt, Julie et al. (2014) Junctional cleft [Ca²?]i measurements using novel cleft-targeted Ca²? sensors. Circ Res 115:339-47
Despa, Sanda; Sharma, Savita; Harris, Todd R et al. (2014) Cardioprotection by controlling hyperamylinemia in a ""humanized"" diabetic rat model. J Am Heart Assoc 3:
Despa, Sanda; Bers, Donald M (2013) Na? transport in the normal and failing heart - remember the balance. J Mol Cell Cardiol 61:2-10

Showing the most recent 10 out of 15 publications