Stem cell injections into the heart are actively being pursued as a potential therapy for myocardial infarction and heart failure. While the ongoing trials with adult-derived stem cells show moderate clinical benefits, significant progress in the field is expected to arise from the use of cardiomyocytes derived from induced pluripotent stem cells. Despite great promise, eventual clinical use of pluripotent stem cell-derived cardiomyocytes faces a number of challenges that need to be resolved including key issues with inadequate cell maturation, phenotypic heterogeneity, arrhythmogenesis, low viability after implantation, and scale-up. Therefore, in this project we aim to establish a novel approach for cardiac cell and gene therapy that does not rely on the use of stem cells. Instead we propose to employ in vitro or in situ genetic engineering of fibroblasts into electrically active cells with customizable electrical phenotype that can couple with surrounding cardiomyocytes and improve their electrical and contractile function. Specifically, in Aim 1 we will utilize minimum st of genetic manipulations to rapidly and efficiently convert adult human fibroblasts into a readily expandable and homogeneous source of excitable cells that autonomously fire and conduct action potentials.
In Aim 2, engineered fibroblasts with select electrophysiological phenotypes will be characterized for their functional interactions with neonatal rat cardiomyocytes in well-controlled in vitro co- culture systems.
In Aim 3, we will establish if contractile function of infarcted rat hearts can be improved by implantation of engineered excitable fibroblasts or retroviral conversion of endogenous fibroblasts into electrically active cells. In addition to abov experimental studies, we will utilize computer simulations to facilitate genetic engineering of excitable cells and enhance mechanistic understanding of their functional interactions with native cardiomyocytes in vitro and in vivo. We believe that the proposed genetic and tissue engineering approach will provide strong foundation for the future experimental and clinical use of engineered fibroblasts in cell- and gene-based therapies for cardiac infarction and arrhythmias.

Public Health Relevance

This project aims to develop and validate a new bioengineering approach for treatment of myocardial infarction and heart failure whereby genetic engineering techniques are utilized to rapidly and efficiently convert unexcitable primary fibroblasts into electrically excitable and actively conducting cells. Transplantation of tissue-engineered patches made of engineered fibroblasts and gene therapy targeted to endogenous fibroblasts will be explored for treatment of post-infarction heart disease.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
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Special Emphasis Panel (ZRG1)
Program Officer
Lee, Albert
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Duke University
Biomedical Engineering
Biomed Engr/Col Engr/Engr Sta
United States
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Gokhale, Tanmay A; Asfour, Huda; Verma, Shravan et al. (2018) Microheterogeneity-induced conduction slowing and wavefront collisions govern macroscopic conduction behavior: A computational and experimental study. PLoS Comput Biol 14:e1006276
Jackman, Christopher P; Ganapathi, Asvin M; Asfour, Huda et al. (2018) Engineered cardiac tissue patch maintains structural and electrical properties after epicardial implantation. Biomaterials 159:48-58
Nguyen, Hung X; Kirkton, Robert D; Bursac, Nenad (2018) Generation and customization of biosynthetic excitable tissues for electrophysiological studies and cell-based therapies. Nat Protoc 13:927-945
Bassat, Elad; Mutlak, Yara Eid; Genzelinakh, Alex et al. (2017) The extracellular matrix protein agrin promotes heart regeneration in mice. Nature 547:179-184
Li, Yanzhen; Asfour, Huda; Bursac, Nenad (2017) Age-dependent functional crosstalk between cardiac fibroblasts and cardiomyocytes in a 3D engineered cardiac tissue. Acta Biomater 55:120-130
Yanamandala, Mounica; Zhu, Wuqiang; Garry, Daniel J et al. (2017) Overcoming the Roadblocks to Cardiac Cell Therapy Using Tissue Engineering. J Am Coll Cardiol 70:766-775
Liau, Brian; Jackman, Christopher P; Li, Yanzhen et al. (2017) Developmental stage-dependent effects of cardiac fibroblasts on function of stem cell-derived engineered cardiac tissues. Sci Rep 7:42290
Shadrin, Ilya Y; Allen, Brian W; Qian, Ying et al. (2017) Cardiopatch platform enables maturation and scale-up of human pluripotent stem cell-derived engineered heart tissues. Nat Commun 8:1825
Pomeroy, Jordan E; Nguyen, Hung X; Hoffman, Brenton D et al. (2017) Genetically Encoded Photoactuators and Photosensors for Characterization and Manipulation of Pluripotent Stem Cells. Theranostics 7:3539-3558
Gokhale, Tanmay A; Kim, Jong M; Kirkton, Robert D et al. (2017) Modeling an Excitable Biosynthetic Tissue with Inherent Variability for Paired Computational-Experimental Studies. PLoS Comput Biol 13:e1005342

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