One of the most important potential uses of embryonic stem cells and induced pluripotent stem cell research is in the development of disease models. Although many diseases can be modeled in mice, others are difficult or impossible to adequately model in non-human organisms for a variety of reasons. In the retina, for example, there are mouse models for many inherited degenerations, but these have been more difficult to generate for diseases like macular degeneration and many forms of Usher's disease. Therefore, disease models from human patient iPS cells would be of great utility in retinal research. In the past six years, our group, and others, have developed protocols for efficient production of retinal cells from human ESCs and iPSCs. The early stages of retinal development are extremely well recapitulated by our protocol of directed differentiation of hESC cells;however, the cells fail to attain the level of expression of markers that we observe in the late stages of human fetal or postnatal development, even with prolonged culture periods. Therefore, a significant challenge for creation of disease models from iPSCs will be to better understand and promote the progression of the cultured cells to mature stages of retina. The mechanisms that control the developmental timing of retinal progenitor cells are not clear;however, recent evidence from our lab shows a key role for miRNAs. Specifically, we found that the loss of Dicer in retinal progenitor cells leads to their failure to progress from the "early" state to the "late" state. We therefore hypothesize (1) that miRNAs regulate developmental timing in retinal progenitor cells and (2) that misregulation of the heterochronic pathway in ESC-derived retinal cells accounts for their failure to progress in vitro at the same rate as in vivo. We propose to test these hypotheses with the following specific aims.
Aim 1 : Determine whether the expression of specific miRNAs (and the genes they regulate) correlates with the progression of retinal progenitors from the early to late stage.
Aim 2. Determine whether the developmental progression of mouse ESC/iPSC-derived retinal progenitors is controlled by stage-specific progenitor miRNAs.
Aim 3. Determine whether the developmental progression of human ESC derived retinal progenitors can be accelerated by stage-specific miRNAs. The results of these studies will enable us to better control the development of hESC-derived retinal cells and produce more appropriate models of retinal disease.

Public Health Relevance

The loss of sight from degenerative diseases of the retina is a major human health problem. Millions of Americans are affected with these diseases and since many are age-related, the number of affected individuals is expected to increase as the population ages. Our research is aimed at determining whether stem cells can be used to treat these diseases, both through cell therapy and disease modeling with induced pluripotent cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
2P01GM081619-06
Application #
8460658
Study Section
Special Emphasis Panel (ZRG1-OBT-A (40))
Project Start
2012-12-17
Project End
2017-11-30
Budget Start
2012-12-17
Budget End
2013-11-30
Support Year
6
Fiscal Year
2013
Total Cost
$278,112
Indirect Cost
$117,740
Name
University of Washington
Department
Type
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Ellen Kreipke, Rebecca; Wang, Yuliang; Miklas, Jason Wayne et al. (2016) Metabolic remodeling in early development and cardiomyocyte maturation. Semin Cell Dev Biol 52:84-92
Hofsteen, Peter; Robitaille, Aaron M; Chapman, Daniel Patrick et al. (2016) Quantitative proteomics identify DAB2 as a cardiac developmental regulator that inhibits WNT/β-catenin signaling. Proc Natl Acad Sci U S A 113:1002-7
Ruan, Jia-Ling; Tulloch, Nathaniel L; Razumova, Maria V et al. (2016) Mechanical Stress Conditioning and Electrical Stimulation Promote Contractility and Force Maturation of Induced Pluripotent Stem Cell-Derived Human Cardiac Tissue. Circulation 134:1557-1567
Pioner, Josè Manuel; Racca, Alice W; Klaiman, Jordan M et al. (2016) Isolation and Mechanical Measurements of Myofibrils from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Stem Cell Reports 6:885-96
Xu, Zhuojin; Robitaille, Aaron M; Berndt, Jason D et al. (2016) Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells. Proc Natl Acad Sci U S A 113:E6382-E6390
Roberts, Meredith A; Tran, Dominic; Coulombe, Kareen L K et al. (2016) Stromal Cells in Dense Collagen Promote Cardiomyocyte and Microvascular Patterning in Engineered Human Heart Tissue. Tissue Eng Part A 22:633-44
Kolwicz Jr, Stephen C; Odom, Guy L; Nowakowski, Sarah G et al. (2016) AAV6-mediated Cardiac-specific Overexpression of Ribonucleotide Reductase Enhances Myocardial Contractility. Mol Ther 24:240-50
Fernandes, Sarah; Chong, James J H; Paige, Sharon L et al. (2015) Comparison of Human Embryonic Stem Cell-Derived Cardiomyocytes, Cardiovascular Progenitors, and Bone Marrow Mononuclear Cells for Cardiac Repair. Stem Cell Reports 5:753-62
Xing, Yalan; Su, Tin Tin; Ruohola-Baker, Hannele (2015) Tie-mediated signal from apoptotic cells protects stem cells in Drosophila melanogaster. Nat Commun 6:7058
Brzezinski, Joseph A; Reh, Thomas A (2015) Photoreceptor cell fate specification in vertebrates. Development 142:3263-73

Showing the most recent 10 out of 95 publications