This application addresses broad Challenge Area (14) Stem Cells and specific Challenge Topic, 14-DK-101 Induced pluripotent stem cells--cellular and humanized mouse models of disease. The liver is the main site of intermediary metabolism of amino acids, carbohydrates, lipids and xenobiotics. Metabolic pathways, especially those of xenobiotic compounds, are highly species specific and can vary even between individuals of the same species. Furthermore, many of the therapeutic strategies - especially gene therapy - envisioned for the treatment of metabolic disorders would be optimally tested in human hepatocytes. We have developed a highly efficient and robust method for the repopulation of murine liver with human hepatocytes and replacement levels of >90% can be achieved. We also have developed the technique to convert human skin fibroblasts into hepatocytes in vitro. Here, we propose to utilize a combination of these methods to produce murine models of human hepatic metabolism. Specifically, we plan to model common inborn errors of lipid metabolism as well as the most common genetic variants of hepatic drug metabolism.

Public Health Relevance

Here we propose to generate mice harboring humanized livers that can be used to study and treat inborn errors of human liver function. This will be achieved by first generating induced pluripotent stem cells from skin cells isolated from patients with hepatic metabolic disorders. These cells will be differentiated into hepatocytes that will be used to repopulate the livers of FRG mice. We have shown that this mouse allows replacement of 90% of its liver with human hepatocytes. These mice will allow the study of metabolic deficiencies, analyses of drug metabolism, screens for novel pharmaceutical treatments of metabolic diseases, as well as form the basis for testing stem cell based gene therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
NIH Challenge Grants and Partnerships Program (RC1)
Project #
5RC1DK087377-02
Application #
7936998
Study Section
Special Emphasis Panel (ZRG1-DKUS-A (58))
Program Officer
Serrano, Jose
Project Start
2009-09-30
Project End
2011-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
2
Fiscal Year
2010
Total Cost
$498,778
Indirect Cost
Name
Medical College of Wisconsin
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
937639060
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Jing, Ran; Corbett, James L; Cai, Jun et al. (2018) A Screen Using iPSC-Derived Hepatocytes Reveals NAD+ as a Potential Treatment for mtDNA Depletion Syndrome. Cell Rep 25:1469-1484.e5
Cayo, Max A; Mallanna, Sunil K; Di Furio, Francesca et al. (2017) A Drug Screen using Human iPSC-Derived Hepatocyte-like Cells Reveals Cardiac Glycosides as a Potential Treatment for Hypercholesterolemia. Cell Stem Cell 20:478-489.e5
Pournasr, Behshad; Duncan, Stephen A (2017) Modeling Inborn Errors of Hepatic Metabolism Using Induced Pluripotent Stem Cells. Arterioscler Thromb Vasc Biol 37:1994-1999
Fisher, J B; Pulakanti, K; Rao, S et al. (2017) GATA6 is essential for endoderm formation from human pluripotent stem cells. Biol Open 6:1084-1095
Nagaoka, Masato; Kobayashi, Motohiro; Kawai, Chie et al. (2015) Design of a Vitronectin-Based Recombinant Protein as a Defined Substrate for Differentiation of Human Pluripotent Stem Cells into Hepatocyte-Like Cells. PLoS One 10:e0136350
Mallanna, Sunil K; Duncan, Stephen A (2013) Differentiation of hepatocytes from pluripotent stem cells. Curr Protoc Stem Cell Biol 26:Unit 1G.4.
Shan, Jing; Schwartz, Robert E; Ross, Nathan T et al. (2013) Identification of small molecules for human hepatocyte expansion and iPS differentiation. Nat Chem Biol 9:514-20
Cayo, Max A; Cai, Jun; DeLaForest, Ann et al. (2012) JD induced pluripotent stem cell-derived hepatocytes faithfully recapitulate the pathophysiology of familial hypercholesterolemia. Hepatology 56:2163-71
DeLaForest, Ann; Nagaoka, Masato; Si-Tayeb, Karim et al. (2011) HNF4A is essential for specification of hepatic progenitors from human pluripotent stem cells. Development 138:4143-53
Si-Tayeb, Karim; Noto, Fallon K; Sepac, Ana et al. (2010) Generation of human induced pluripotent stem cells by simple transient transfection of plasmid DNA encoding reprogramming factors. BMC Dev Biol 10:81

Showing the most recent 10 out of 12 publications