Deriving hepatocytes from disease specific iPS to treat metabolic liver disorders

Summary

Principal Investigator: Namita Roy-Chowdhury
Abstract: DESCRIPTION (provided by applicant): Hepatocyte transplantation is being developed as a minimally invasive alternative to liver transplantation for patients with liver failure or inherite metabolic disorders. Major hurdles to the broad application of this promising therapeutic modality are the scarcity of high quality donor organs and the need for prolonged immunosuppression to prevent allograft rejection. This research proposal is to test the hypothesis that induced pluripotent stem cells (iPSC) derived from the skin of patients with Crigler-Najjar syndrome type 1 (CN1), who have severe lifelong jaundice due to uridinediphosphoglucuronate glucuronosltransferase-1 (UGT1A1) deficiency can be corrected genetically, and then manipulated in culture to generate hepatocytes expressing active UGT1A1. We further hypothesize that appropriate host preparation will permit sufficient repopulation of the livers of the Gunn rat model of CN1 to ameliorate hyperbilirubinemia. The specific aim 1 is (a) Generating and characterizing iPS cells from skin fibroblasts from normal subjects and CN1 patients, (b) correcting the genetic defect by homologous recombination at a genomic "safe harbor", using zinc-finger nuclease or TALE-nuclease technology, and (c) differentiating the genetically corrected iPSCs into hepatocyte-like cells (iHep) in culture. The gene expression profile and metabolic function of the derived hepatocytes will be compared with those of adult and fetal human hepatocytes. Specific aim 2 is (a) Transplanting the iHeps into the liver of Gunn rats following appropriate host preparation, and evaluating the metabolic effect of the transplantation. Gunn rats will be treated with a tacrolimus-based immunosuppressive regimen that has been shown to prevent the rejection of xenografted human hepatocytes in our laboratory. The recipients will be subjected to preparative hepatic irradiation and mitotic stimulation by a single injection of an adenovector, expressing hepatocyte growth factor to induce repopulation of the liver with the transplanted hepatocytes. Serum bilirubin levels will be followed, the extent of liver repopulation will be determined and biliary excretion of bilirubin glucuronides will be analyzed by high-pressure liquid chromatography. (b) Finally, a group of recipient rats and immunodeficient mice will be observed life-long to evaluate the tumorigenic potential of the transplanted iPSC-derived hepatocytes. Successful completion of this project will provide a renewable source of autologous hepatocytes that, after additional modification and scale up, could be translated into cell transplantation-based treatment of CN1 and many other inherited liver diseases.
Funding Period: 2013-08-19 - 2017-07-31
more information: NIH RePORT

Detail Information

Research Grants30

  1. Neural Mechanisms of Itch
    ROBERT H LA MOTTE; Fiscal Year: 2013
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  2. Immune Cells in Atherosclerosis and Vascular Disease
    Catherine C Hedrick; Fiscal Year: 2013
    ..Each project will utilize the Human Core to study immune cells from human subjects to establish functional links between candidate genes of interest and immune cell function in atherogenesis. ..
  3. iPS-derived hepatocytes as a model of familial hypercholesterolemia
    Max A Cayo; Fiscal Year: 2013
    ..abstract_text> ..
  4. Genomics of Kidney Transplantation Admin
    Arthur J Matas; Fiscal Year: 2013
    ..The Administrative Core will also facilitate interaction between Sites, Cores, and Projects. ..
  5. T CELL MEMORY TO PATHOGENS: GENERATION AND FUNCTION
    Susan L Swain; Fiscal Year: 2013
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  6. Center for Molecular Imaging Research at MGH/HMS
    Ralph Weissleder; Fiscal Year: 2013
    ....
  7. University of Maryland Greenebaum Cancer Center Support Grant
    Kevin J Cullen; Fiscal Year: 2013
    ..Reflecting our remarkable and continued growth, UMGCC seeks to renew its CCSG to enhance and expand its efforts in high-quality and clinically relevant cancer research. ..
  8. Testing the Therapeutic Potential of iPS Cells for Inherited Skin Diseases
    Jakub Tolar; Fiscal Year: 2013
    ..If the studies outlined in this application are successful, these pre-clinical data will pave the way for approval of iPSC-based clinical trials for not only other forms of EB, but also other inherited skin disorders. ..
  9. Human Pharmacogenetics and Human Liver Regeneration
    GARY A PELTZ; Fiscal Year: 2013
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  10. Cardiac Myosin Binding Protein-C: Structure, Function, and Regulation
    David M Warshaw; Fiscal Year: 2013
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  11. Transplantation of Endothelial Cells
    Sanjeev Gupta; Fiscal Year: 2013
    ..Fundamental knowledge in respect to the properties and uses of human endothelial cells will advance insights in liver biology and liver cell therapy, which will be of enormous value for improving human health. ..
  12. Center for Gene Therapy of Cystic Firbosis
    John F Engelhardt; Fiscal Year: 2013
    ..These efforts have led to numerous basic and applied research findings that have enhanced the utility of gene therapies to both study and treat genetic diseases. ..
  13. Blood Pressure Regulation: Novel Roles for the Kidney
    Pablo A Ortiz; Fiscal Year: 2013
    ..Thus it will accelerate acquisition of knowledge of the novel mechanisms by which the kidney regulates blood pressure, and may provide new targets for anti-hypertensive drugs. ..
  14. Designing a Microenvironment Niche for Liver-Specific Differentiation of hESCs
    Alexander Revzin; Fiscal Year: 2013
    ..Because our platform will be miniature in scale, we will be able to complete many experiments at once to define an optimal liver-specific microenvironment without needing an abundance of cells or proteins. ..
  15. Engineering a Functional Liver Graft for Treatment of End Stage Liver Disease
    DENVER MICHAEL FAULK; Fiscal Year: 2013
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  16. STEM CELL GENE THERAPY FOR HEMOGLOBINOPATHIES
    George Stamatoyannopoulos; Fiscal Year: 2013
    ..The focus of this Program Project, Gene Therapy, can provide a new paradigm for the treatment of these hemoglobinopathies as well as for other blood diseases. (End of Abstract) ..
  17. BIOLOGY OF NEUROENDOCRINE PEPTIDES
    Marc R Montminy; Fiscal Year: 2013
    ..Specifying the contributions of the CRF family of ligands and receptors to the maintenance of homeostasis and to stress-linked allostasis may improve our ability to manage diseases, including mood and metabolic disorders ..
  18. Regulating fibrosis and muscle growth in the muscular dystrophies
    Elizabeth M McNally; Fiscal Year: 2013
    ..Core B will provide histopathological assessment of muscular dystrophy after genetic manipulation and treatments, and Core C will perform functional analysis in vivo and provide support to Core B. ..
  19. Synaptic Function: Effects of the Nerve Injury, Repair, and Altered Activity
    Timothy C Cope; Fiscal Year: 2013
    ..The Resume and Summary of Discussion above summarizes the final outcome of the group discussion. OVERALL PROGRAM EVALUATION ..