Testing the Therapeutic Potential of iPS Cells for Inherited Skin Diseases

Summary

Principal Investigator: Jakub Tolar
Abstract: DESCRIPTION (provided by applicant): Epidermolysis bullosa (EB) is a group of rare inherited skin blistering diseases that result in severe blistering and scaring. Some of the variants of EB sentence those afflicted to a life of severe pain and disability and even early death. Although the genetic defects that cause these devastating diseases have been known for almost 2 decades, current therapy for EB is limited to wound care. Since the epidermis is continuously renewed by stem cells in the proliferative basal layer, a permanent corrective therapy for EB must target the stem cell population. To date, no one has reported the successful use of homologous recombination technology to correct a defective gene in human keratinocyte stem cells isolated from an EB patient. The only successful clinical trial for an inherited skin blistering disease utilized a retroviral vector to restore expression of the missing protein, and that trial was placed on hold because of safety concerns with the use of retroviral vectors. For this reason, we believe that alternative strategies must be explored. Therefore, we are proposing to develop stem-cell based therapies for EB using autologous induced pluripotent stem cells (iPSC) derived from skin cells harvested from the same EB patient. The generation of patient-specific iPSC would not only potentially avoid the complication of immune rejection, but also provide a source of rejuvenated adult stem cells that are most likely exhausted as a result of unsuccessful attempts to repair blistered tissues. Prior to testing an iPSC-based stem cell therapy for EB in humans, it is desirable to utilize a pre-clinical animal model to determine the safety and efficacy of these approaches. We have previously generated an inducible mouse model that mimics the most severe form of epidermolysis bullosa simplex, Dowling-Meara (EBS-DM), at the genetic level. Using this model, we have obtained very compelling data documenting that EBS keratinocyte stem cells exhibit a growth disadvantage compared to wild-type keratinocyte stem cells. These results suggest that EBS is an example of an inherited skin disease where there would be a "natural selection" for genetically corrected iPSC-derived keratinocytes if they were seeded into areas prone to blistering. For this reason, and the fact that genetically corrected iPSC-derived keratinocytes would not be rejected by the patient's immune system, since EBS is a dominant form of EB, we believe that EBS is an ideal model for generating "proof of concept" data supporting the use of iPSC for the treatment of EB. Patients with EBS-DM develop lesions in the skin and the oral mucosal epithelia;therefore, both local and systemic therapy will be required to treat these patients. Dr. Jakub Tolar was the first to perform clinical trials using systemically delivered allogeneic bone marrow (BM)-derived cells in the treatment of RDEB patients, and for that reason, he was recruited as a PI on this multi-PI application. Although Dr. Tolar's initial clinical trials were very promising, there are safety concerns with allogeneic transplants, such as toxicity to chemotherapy required for conditioning before transplantation, and susceptibility to infections due to the necessity for immunosuppressive therapy to prevent rejection of allogeneic transplants. Unfortunately, 2 of the 7 RDEB children included in this initial trial died due to these complications. To avoid the complications of allogeneic transplants, we are proposing to generate iPSC from EBS-DM patients, and use zinc-finger nuclease (ZFN)-mediated genome editing to inactivate the mutant keratin 14 (KRT14) allele, which is defective in the majority of EBS-DM patients. We are proposing to use these corrected iPSC to generate keratinocytes to repair the skin by grafting, and mesenchymal cells to systemically repair lesions in the oral cavity. We have made substantial progress in overcoming some of the obstacles that will be required before iPSC can be used safely in the clinic, such as developing a non-viral method for reprogramming and efficient protocols for differentiating iPSC into keratinocytes and mesenchymal cells, but other safety issues remain. In this application, we are proposing to use novel models to address the remaining issues of histocompatibility, tumorigenicity, and genetic stability of iPSC-derived cells We have also designed a ZFN-mediated strategy to inactivate a 'hot spot mutation" responsible for ~70% of the EBS-DM cases. 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.
Funding Period: 2012-09-17 - 2017-08-31
more information: NIH RePORT

Top Publications

  1. pmc Allogeneic blood and bone marrow cells for the treatment of severe epidermolysis bullosa: repair of the extracellular matrix
    Jakub Tolar
    Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    Lancet 382:1214-23. 2013
  2. pmc Patient-specific naturally gene-reverted induced pluripotent stem cells in recessive dystrophic epidermolysis bullosa
    Jakub Tolar
    1 Department of Pediatrics, Blood and Marrow Transplant, Medical School, University of Minnesota, Minneapolis, Minnesota, USA 2 Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA
    J Invest Dermatol 134:1246-54. 2014
  3. pmc Differentiation of human induced pluripotent stem cells into a keratinocyte lineage
    Igor Kogut
    Charles C Gates Center for Regenerative Medicine and Stem Cell Biology, University of Colorado, Anschutz Medical Campus, Mail Stop 8320, 6511, Aurora, CO, 80045, USA
    Methods Mol Biol 1195:1-12. 2014

Detail Information

Publications3

  1. pmc Allogeneic blood and bone marrow cells for the treatment of severe epidermolysis bullosa: repair of the extracellular matrix
    Jakub Tolar
    Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    Lancet 382:1214-23. 2013
    ..Future strategies will be built on the lessons learned from these first transplant studies. ..
  2. pmc Patient-specific naturally gene-reverted induced pluripotent stem cells in recessive dystrophic epidermolysis bullosa
    Jakub Tolar
    1 Department of Pediatrics, Blood and Marrow Transplant, Medical School, University of Minnesota, Minneapolis, Minnesota, USA 2 Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA
    J Invest Dermatol 134:1246-54. 2014
    ..We believe this approach should be the starting point for autologous cellular therapies using 'natural' gene therapy in RDEB and other diseases...
  3. pmc Differentiation of human induced pluripotent stem cells into a keratinocyte lineage
    Igor Kogut
    Charles C Gates Center for Regenerative Medicine and Stem Cell Biology, University of Colorado, Anschutz Medical Campus, Mail Stop 8320, 6511, Aurora, CO, 80045, USA
    Methods Mol Biol 1195:1-12. 2014
    ....

Research Grants30

  1. Keratin Gene Targeting for the Treatment of Epidermolysis Bullosa
    Daniel G Miller; Fiscal Year: 2013
    ..There is currently no effective therapy for preventing these blisters. This proposal describes experiments designed to test gene therapy strategies for the treatment of epidermolysis bullosa simplex. ..
  2. GENE THERAPY USING HEMATOPOIETIC STEM CELLS
    Donald B Kohn; Fiscal Year: 2013
    ..The Project and Core leaders have complementary expertise in the relevant areas of experimental hematology, gene therapy, immunology, and signal transduction and have a long-standing record of interactive collaborations. ..
  3. Molecular and Architectural Mechanisms of Reprogramming to Pluripotency
    Kathrin Plath; Fiscal Year: 2013
    ..The plan provides unique experimental synergies that address the objectives of the funding announcement. ..
  4. Intellectual and Development Disabilities Research Center
    Marc Yudkoff; Fiscal Year: 2013
    ..5 million from NICHD). The Center includes an excess of 70 Penn faculty at 15 departments at the Schools of Medicine, Veterinary Medicine, Nursing, the Wistar Institute, and the College of Arts and Sciences. ..
  5. Gene Transfer for Recessive Dystrophic Epidermolysis Bullosa
    MATT PETER MARINKOVICH; Fiscal Year: 2013
    ..We plan to correct the affected person's own skin cells and transplant their own cells back onto their skin. Effective gene therapy in this skin disease will assist in development of gene therapy for other genetic diseases. ..
  6. THE UCLA CENTER FOR IN VIVO IMAGING IN CANCER BIOLOGY
    Harvey R Herschman; Fiscal Year: 2013
    ....
  7. Induced pluripotent stem cell therapy for lipodystrophy
    Susan M Majka; Fiscal Year: 2013
    ..Additionally, these studies will generate important resources and reagents that will be of vast interest to both the scientific and medical communities. ..
  8. M. D. Anderson Cancer Center SPORE in Multiple Myeloma
    ROBERT ZYGMUNT ORLOWSKI; Fiscal Year: 2013
    ..abstract_text> ..
  9. 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. ..
  10. Jules Stein Eye Institute Core Grant for Vision Research
    Wayne L Hubbell; Fiscal Year: 2013
    ..Support in the form of the Core grant is requested to maintain these Modules through instrument service contracts, and to provide necessary personnel support to assist and train users and provide routine maintenance. ..
  11. Center for Neuroplasticity at the University of Puerto Rico
    Steven N Treistman; Fiscal Year: 2013
    ..This UPR COBRE Center should define pathways and benchmarks for basic and translational research across the UPR system for the next decades. ..
  12. Signaling in Inflammation, Stress, and Tumorigenesis
    GEORGE ROBERT STARK; Fiscal Year: 2013
    ..abstract_text> ..
  13. 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. ..
  14. 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) ..
  15. Core Center for Musculoskeletal Disorders
    Louis J Soslowsky; Fiscal Year: 2013
    ..abstract_text> ..
  16. Targeted Correction of Dominant Mutations of Type I Collagen Causing Severe OI
    David W Rowe; Fiscal Year: 2013
    ....
  17. Neural Mechanisms of Itch
    ROBERT H LA MOTTE; Fiscal Year: 2013
    ..abstract_text> ..
  18. Columbia University Medical Center Skin Disease Research Center (CUMCSDRC)
    DAVID RINSEY BICKERS; Fiscal Year: 2013
    ..abstract_text> ..
  19. Skin-targeted Cell Therapy for Recessive Dystrophic Epidermolysis Bullosa
    Jakub Tolar; Fiscal Year: 2013
    ....
  20. Muscular Dystrophy Center Core Laboratories
    James M Ervasti; Fiscal Year: 2013
    ..4) To increase institutional, regional and national awareness of UMN-MDCenter and CCMBM to enhance provision of institutional resources commensurate with the potential of this program. ..