Premium
S100A1 gene transfer: a strategy to strengthen engineered cardiac grafts
Author(s) -
Remppis Andrew,
Pleger Sven T.,
Most Patrick,
Lindenkamp Jan,
Ehlermann Philipp,
Schweda Christopher,
Löffler Eva,
Weichenhan Dieter,
Zimmermann Wolfram,
Eschenhagen Thomas,
Koch Walter J.,
Katus Hugo A.
Publication year - 2004
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.513
Subject(s) - contractility , transfection , isometric exercise , extracellular , western blot , calcium , microbiology and biotechnology , medicine , basal (medicine) , chemistry , gene transfer , genetic enhancement , gene , andrology , endocrinology , biology , biochemistry , insulin
Background Cardiac tissue replacement therapy, although a promising novel approach for the potential treatment of heart failure, still suffers from insufficient contractile support to the failing myocardium. Here, we explore a strategy to improve contractile properties of engineered heart tissue (EHT) by S100A1 gene transfer. Methods EHTs were generated from neonatal rat cardiomyocytes and transfected (MOI 10 PFU) with the S100A1 adenovirus (AdvS100A1, n = 25) while an adenovirus devoid of the S100A1 cDNA served as a control (AdvGFP, n = 30). Contractile properties of transfected EHTs were measured 7 days following gene transfer. Results Western blot analysis confirmed a 8.7 ± 3.6‐fold S100A1 protein overexpression in AdvS100A1‐transfected EHTs ( n = 4; P < 0.01) that increased maximal isometric force (mN; AdvGFP 0.175 ± 0.03 vs. AdvS100A1 0.47 ± 0.06; P < 0.05) at 0.4 mmol/L extracellular calcium concentration [Ca 2+ ] e . In addition, S100A1 overexpression enhanced both maximal Ca 2+ ‐stimulated force generation (+81%; P < 0.05) and Ca 2+ ‐sensitivity of EHTs (EC50% [Ca 2+ ] e mM; AdvGFP 0.33 ± 0.04 vs. AdvS100A1 0.21 ± 0.0022; P < 0.05). The S100A1‐mediated gain in basal graft contractility was preserved throughout a series of isoproterenol interventions (10 −9 to 10 −6 M). Physiological properties of EHTs resembling intact heart preparations were preserved. Conclusions S100A1 gene transfer in EHT is feasible and augments contractile performance, while characteristic physiological features of EHT remain unchanged. Thus, specific genetic manipulation of tissue constructs prior to implantation should be part of an improved tissue replacement strategy in heart failure. Copyright © 2004 John Wiley & Sons, Ltd.