Treatments for cardiovascular diseases are significant unmet needs in the global medical community. We propose to develop in vitro models of diseased cardiac tissues by using precisely controlled artificial matrix preparations. The principal objective of this project is to establish an in vitro model of human cardiac tissue based on the reconstitution of synthetic models of the human ventricular myocardium with populations of patient specific human induced pluripotent stem (hiPS) cell-derived cardiomyocytes (hiPS-CMs). For this application we have chosen to focus on a single """"""""patient-specific"""""""" disease, long QT syndrome (LQTS), as a basis for proof-of-principle of our methodology and workflow. Prolongation of the QT interval, the electrical manifestation of cardiac ventricular repolarization, is a major cause of cardiac arrhythmias and sudden death. Thus a LQTS """"""""patient-specific"""""""" physiologically functioning 3D model of heart tissue would be a significant advancement for understanding, studying, and developing new strategies for treating cardiac arrhythmias and other cardiovascular diseases. We propose the following specifics aims to generate a human cardiac 3D tissue model.
Aim 1. To optimize a directed differentiation method to obtain a consistent high yield (>75%) population of human CMs derived from either healthy hiPS or hiPS cells harboring gene mutations of LQTS, a potentially lethal mutation.
Aim 2. To fabricate precisely defined 3D filamentous matrices that organize the structure of healthy hiPS-CMs into a 3D in vitro model of the human cardiac tissue. To assess the functional behavior of the model by examining its electrical and mechanical activity.
Aim 3. To organize the structure of LQTS-hiPS-CMs into a 3D in vitro model of the human myocardium. To assess the functional behavior of the """"""""diseased tissue"""""""" model by examining its electrical and mechanical activity, and response to pharmacological agents.

Public Health Relevance

This application will focus on developing a patient-specific physiologically functioning three-dimensional model of cardiac tissue. This tissue model represents a significant advancement for understanding, studying, and developing new strategies for treating cardiovascular disease. This project will focus on induced pluripotent stem (iPS) cells, also known as stem cells from skin cells, to form the cardiac tissues that can be widely used by the research community.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL108677-04
Application #
8676918
Study Section
Special Emphasis Panel (ZHL1)
Program Officer
Lundberg, Martha
Project Start
2011-09-05
Project End
2015-05-31
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
4
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Engineering (All Types)
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Kime, Cody; Mandegar, Mohammad A; Srivastava, Deepak et al. (2016) Efficient CRISPR/Cas9-Based Genome Engineering in Human Pluripotent Stem Cells. Curr Protoc Hum Genet 88:Unit 21.4
Mandegar, Mohammad A; Huebsch, Nathaniel; Frolov, Ekaterina B et al. (2016) CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Cell Stem Cell 18:541-53
Miyaoka, Yuichiro; Berman, Jennifer R; Cooper, Samantha B et al. (2016) Systematic quantification of HDR and NHEJ reveals effects of locus, nuclease, and cell type on genome-editing. Sci Rep 6:23549
Huebsch, Nathaniel; Loskill, Peter; Deveshwar, Nikhil et al. (2016) Miniaturized iPS-Cell-Derived Cardiac Muscles for Physiologically Relevant Drug Response Analyses. Sci Rep 6:24726
Ma, Zhen; Wang, Jason; Loskill, Peter et al. (2015) Self-organizing human cardiac microchambers mediated by geometric confinement. Nat Commun 6:7413
Huebsch, Nathaniel; Loskill, Peter; Mandegar, Mohammad A et al. (2015) Automated Video-Based Analysis of Contractility and Calcium Flux in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Cultured over Different Spatial Scales. Tissue Eng Part C Methods 21:467-79
Nobuta, Hiroko; Cilio, Maria Roberta; Danhaive, Olivier et al. (2015) Dysregulation of locus coeruleus development in congenital central hypoventilation syndrome. Acta Neuropathol 130:171-83
Ma, Zhen; Koo, Sangmo; Finnegan, Micaela A et al. (2014) Three-dimensional filamentous human diseased cardiac tissue model. Biomaterials 35:1367-77
Spencer, C Ian; Baba, Shiro; Nakamura, Kenta et al. (2014) Calcium transients closely reflect prolonged action potentials in iPSC models of inherited cardiac arrhythmia. Stem Cell Reports 3:269-81
Miyaoka, Yuichiro; Chan, Amanda H; Judge, Luke M et al. (2014) Isolation of single-base genome-edited human iPS cells without antibiotic selection. Nat Methods 11:291-3