The long term goal of this application is to create a pure population of cardiac pacemaker cells from an easily accessible autologous cell type, the human hair follicle keratinocyte (HFKT-pacemakers), to characterize their pacemaker mechanism, their ability to integrate into the cardiac syncytium and their potential to function as an in vivo biological pacemaker. If successful in the long term, the health benefit of such an approach will be to substitute for the more than 350,000 electronic pacemakers implanted or reimplanted in patients in the United States each year. The project has four specific aims: (1) to expand the population of induced pluripotent stem cells created from the HFKTs, enhance their differentiation to a cardiac lineage and select for pacemaker myocytes;(2) to characterize the membrane currents in the HFKT- pacemakers as well as their pacemaker function and gene expression profile, and to compare the HFKT- pacemaker to native cardiac primary and secondary pacemakers in vitro;(3) to determine (a) the ability of HFKT pacemakers to couple to adult heart cells from specific locations (atrium or ventricle) in vitro and whether the pacing rate generated is target dependent (b) which connexins HFKT-pacemakers express and the ability of the HFKT-pacemakers to couple to cells expressing fibroblast connexins (a potentially arrhythmogenic situation);(4) to determine in vivo biological pacemaker function generated by placement of HFKT-pacemakers in the canine atrium or ventricle. Our approach will employ 1) novel methods to enhance selection of pacemaker cells, 2) patch clamping to characterize action potential morphology and membrane currents in the isolated pacemaker cells generated, 3) gene chips to determine the pacemaker cells' expression profile, 4) dual whole cell patch clamp, and biochemical and molecular techniques to determine connexin expression and functional cell to cell coupling 5) Injection of the HFKT- pacemakers into the canine atrium or ventricle to determine in vivo pacemaker function. The experiments will be carried out by a team of long term collaborators at Stony Brook University and at the Technion in Israel. The team has extensive expertise in stem cell biology, induced pluripotent stem cells, cardiac pacemaking, patch clamp,, and in vivo studies of biological pacemaker function. Successful execution of the research plan will enhance selection techniques for cardiac cell lineages from IPSCs, characterize the basis of pacemaker activity in the HFKT- pacemakers and determine their effectiveness as an in vivo biological pacemaker. If the HFKT-pacemaker functions well in the canine heart, a future goal would be to advance this novel autologous, cellular approach towards clinical deployment.

Public Health Relevance

Each year 350,000 pacemakers are implanted in American citizens. Although enormously successful they also have drawbacks which limit their effectiveness. The successful development of an autologous biological pacemaker will increase the quality of life for those patients who suffer disorders of cardiac rhythm.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL111401-02
Application #
8423701
Study Section
Electrical Signaling, Ion Transport, and Arrhythmias Study Section (ESTA)
Program Officer
Krull, Holly
Project Start
2012-02-15
Project End
2016-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
2
Fiscal Year
2013
Total Cost
$635,882
Indirect Cost
$186,222
Name
State University New York Stony Brook
Department
Physiology
Type
Schools of Medicine
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Chauveau, Samuel; Anyukhovsky, Evgeny P; Ben-Ari, Meital et al. (2017) Induced Pluripotent Stem Cell-Derived Cardiomyocytes Provide In Vivo Biological Pacemaker Function. Circ Arrhythm Electrophysiol 10:e004508
Ben-Ari, Meital; Naor, Shulamit; Zeevi-Levin, Naama et al. (2016) Developmental changes in electrophysiological characteristics of human-induced pluripotent stem cell-derived cardiomyocytes. Heart Rhythm 13:2379-2387
Kim, Jeremy; Gao, Junyuan; Cohen, Ira S et al. (2015) Angiotensin II Type 1 Receptor-Mediated Electrical Remodeling in Mouse Cardiac Myocytes. PLoS One 10:e0138711
Guo, Yuanjian; Lu, Zhongju; Cohen, Ira Stephen et al. (2015) Development of a universal RNA beacon for exogenous gene detection. Stem Cells Transl Med 4:476-82
Ben-Ari, Meital; Schick, Revital; Barad, Lili et al. (2014) From beat rate variability in induced pluripotent stem cell-derived pacemaker cells to heart rate variability in human subjects. Heart Rhythm 11:1808-1818
Gao, Junyuan; Sun, Xiurong; Potapova, Irina A et al. (2014) Autocrine A2 in the T-system of ventricular myocytes creates transmural gradients in ion transport: a mechanism to match contraction with load? Biophys J 106:2364-74
Chauveau, Samuel; Brink, Peter R; Cohen, Ira S (2014) Stem cell-based biological pacemakers from proof of principle to therapy: a review. Cytotherapy 16:873-80
Krebs, Simone; Gottschalk, Stephen (2014) Cell carriers to attack glioma. Cytotherapy 16:871-2
Barad, Lili; Schick, Revital; Zeevi-Levin, Naama et al. (2014) Human embryonic stem cells vs human induced pluripotent stem cells for cardiac repair. Can J Cardiol 30:1279-87
Clausen, Chris; Valiunas, Virginijus; Brink, Peter R et al. (2013) MATLAB implementation of a dynamic clamp with bandwidth of >125 kHz capable of generating I Na at 37 °C. Pflugers Arch 465:497-507

Showing the most recent 10 out of 14 publications