Defining the role of non-myocytes for cardiomyocyte differentiation and function in an genetically engineered heart muscle model
Final Report Abstract
The aim of the study was to develop cell-specific reporter models (first in the mouse and then in the human) to study the role of myocardial cell-cell interplay for heart muscle formation in vitro. In preliminary work we had already established transgenic embryonic stem cells (ESCs) lines from the mouse, expressing a neomycin resistance (NeoR), green fluorescing protein (GFP), and nuclear localized beta-galactosidase (nLacZ) specifically in cardiomyocytes (aMHC-NIGIL). Building on this work we proposed to use alternative promoter elements to generate ESCs with distinguishable labels in endothelial cells, smooth muscle cells, fibroblasts, and macrophages. Accordingly, we cloned several DNA vectors containing red (NIRIL), yellow (NIYIL), and orange (NIOIL) fluorescing proteins for stable integration in murine ESCs. However, we realized that cell purification and amplification in the genetically modified mouse ESC models is limited, because of suboptimal differentiation and selection protocols. After extensive work to address these limitations, we decided to not aim at specifically selecting cell types for tissue engineering, but focus first on genetically naive fibroblasts isolated from different sources and purified by passaging in culture. With this approach, we could demonstrate that fibroblasts and cardiomyocytes are essential components for the construction of Engineered Heart Muscle (EHM). This was demonstrated for several mouse and human pluripotent stem cell types, including embryonic, induced pluripotent and parthenogenetic stem cells. Finally, in the human model, we developed a transgenic reporter model with RFP and GFP labeled cardiomyocytes and fibroblast-like stroma cells, respectively. This will enable us to gain fundamental insight into processes governing heart muscle assembly and the specific contnbution of fibroblasts in this process. The development of defined conditions for human EHM assembly also serves as the basis for our efforts to clinically translate tissue engineered heart repair, first with embryonic stem cells (1st generation of therapeutic EHM), followed by induced pluripotent stem cells (2nd generation of therapeutic EHM), and parthenogenetic stem cells (3rd generation of therapeutic EHM).
Publications
- (2010) PDGF-BB protects cardiomyocytes from apoptosis and improves contractile function of engineered heart tissue. J Mol Cell Cardiol 48:1316- 1323
Vantler M, Karikkinetti BC, Naito H, Tiburcy M, Didié M, Nose M, Rosenkranz S, Zimmermann WH
- (2011) Terminal Differentiation, Advanced Organotypic Maturation, and Modeling of Hypertrophic Growth in Engineered Heart Tissue. Circ Res. 109:1105-1114
Tiburcy M, Didié M, Boy O, Christalla P, Doeker S, Naito H, Karikkineth BC, El-Armouche A, Grimm M, Nose M, Eschenhagen T, Zieseniss A, Katschinski D, Hamdani N, Linke WA, Yin X, Mayr M, Zimmermann WH
- (2011) Tissue Engineered Myocardium. In Stud Mechanobiol Tissue Eng Biomater. Chapter 41; pp. 111-132; Hrsg: A.R. Boccaccini, S.E. Harding; Springer-Verlag Berlin Heidelberg
Zimmermann WH
- (2012) Cardiac Differentiation of Human Embryonic Stem Cells and their Assembly into Engineered Heart Muscle. Curr Protoc Cell Biol 55:23.8.1-23.8.21
Soong PL, Tiburcy M, Zimmermann WH
- (2012) Engineered Heart Tissue; in Heart Regeneration - Stem Cells and Beyond. Chapter VI Section B; pp. 299-326; Edited by: F. Engel; World Scientific (ISBN: 978-981-4299-80-0)
Eschenhagen T, Zimmermann WH
- (2013) Comparative study of human-induced pluripotent stem cells derived from bone marrow cells, hair keratinocytes, and skin fibroblasts. Eur Heart J. 34:2618-29
Streckfuss-Bömeke K, Wolf F, Azizian A, Stauske M, Tiburcy M, Wagner S, Hübscher D, Dressel R, Chen S, Jende J, Wulf G, Lorenz V, Schön MP, Maier LS, Zimmermann WH, Hasenfuss G, Guan K
(See online at https://doi.org/10.1093/eurheartj/ehs203) - (2013) Emerging Concepts in Myocardial Pharmacoregeneration. Regenerative Medicine; Chapter 25; pp. 637-664. Edited by: G. Steinhoff; Springer
Zelarayan LC, Zafiriou MP, Zimmermann WH
- (2013) Parthenogenetic Stem Cells for Tissue Engineered Heart Repair. J Clin Invest 123:1285-1298
Didié M, Christalla P, Rubart M, Muppala V, Döker S, Unsöld B, El-Armouche A, Rau T, Eschenhagen T, Schwoerer AP, Ehmke H, Schumacher U, Fuchs S, Lange C, Becker A, Tao W, Scherschel JA, Soonpaa MH, Yang T, Lin Q, Zenke M, Han DW, Schöler HR, Rudolph C, Steinemann D, Schlegelberger B, Kattman S, Witty A, Keller G, Field LJ, Zimmermann WH
(See online at https://doi.org/10.1172/JCI66854) - (2013) Pluripotent Stem Celt-Derived Cardiomyocytes for Industrial and Clinical Applications. Modern Biopharmaceuticals; Chapter 3; pp. 57-76; Edited by: J. Knaeblein; Wiley-VCH
Hudson JE, Christalla P, Zimmermann WH
- (2014) Clinical considerations for cardiac tissue engineering. In: Cardiac Regeneration and Repair: Vol. 2, Biomaterials and Tissue Engineering; Edited by: R. Weisel, R.-K. Li; Woodhead Publishing. Kap. 12, S. 299-312
Zimmermann WH
(See online at https://doi.org/10.1533/9780857096715.2.299) - (2014) Collagen-based Engineered Heart Muscle. S. 167-176 in: Radisic M., Black III L. (eds) Cardiac Tissue Engineering. Methods in Molecular Biology (Methods and Protocols), vol 1181. Humana Press, New York, NY. - 978-1-4939-1046-5
Tiburcy M, Meyer T, Soong PL, Zimmermann WH
(See online at https://doi.org/10.1007/978-1-4939-1047-2_15) - (2014) Modeling Myocardial Growth and Hypertrophy in Engineered Heart Muscle. Trends Cardiovasc Med 24:7-13
Tiburcy M, Zimmermann WH
(See online at https://doi.org/10.1016/j.tcm.2013.05.003)
