Recessive dystrophic epidermolysis bullosa (RDEB) is caused by loss-of-function mutations in the collagen type VII (C7) gene (COL7A1), and results in a potentially fatal and terribly debilitating disorder. Individuals with severe generalied RDEB develop blisters and erosions involving large portions of the body surface area, mutilating scarring, joint contractures, and strictures of the esophagus, with opportunistic infections and aggressive squamous cell carcinoma as principal causes of early death. We have shown that hematopoietic stem cell transplantation (HSCT) can increase C7 protein in skin and mucous membranes, and ameliorate many of the disease manifestations. Unfortunately not all mucocutaneous lesions heal, and the toxicities associated with HSCT are significant. Our overarching goal is to solve these problems by establishing a reliable method for delivering C7-expressing cells that home to the dermal-epidermal junction and secrete C7 protein. In order to accomplish this goal and to overcome the limitations of allogeneic HSCT, we will: 1) identify the optimal cellular vector and homing signal for targeting injured skin in RDEB, using human mesenchymal stromal/stem cells and induced pluripotent stem cells (iPSCs), 2) define the efficacy of systemic RDEB gene therapy by C7 gene augmentation, and 3) use nucleases for gene editing of individual mutations, as well as whole regions of the C7 gene. We propose to define the conditions conducive to wound healing in this severe blistering genodermatosis, but the impact of these studies may not be limited to RDEB. The function of C7 depends on extracellular polymerization, which makes RDEB an ideal model for understanding the mechanisms of cross- correction of structural protein deficiency in the extracellular matrix. In addition, wound healing integrates skin extracellular matrix and skin cells into a dynamic system, which in RDEB results in the depletion of skin stem cell niches and, in turn, provides a unique model of skin tissue repair with donor regenerative cells. By using powerful tools for studying and manipulating the information basis of biological systems-engineered restriction enzymes (nucleases), skin- targeting, and induced cell lineage conversion (iPSCs)-we will focus on developing personalized cell therapy for individuals with RDEB with the idea that our findings may have broad implications for understanding other connective tissue extracellular matrix diseases. Our proposal is equally motivated by wanting a better understanding of the biological mechanisms in injured skin and by needing to improve the lives of people with RDEB through maximizing the benefits and reducing the risks of potential novel therapies.

Public Health Relevance

More than 3,000 new cases of genetic inborn errors are found in U.S. newborns every year. The complex and life-long medical care for these patients represents a significant challenge to the health care system. Additionally, a deeper understanding of the basic biology of skin blistering disorders and the proof-of-concept for clinical translation in these relatively rare conditions is likely to have a profound impact on muc more common ones, such as chemical and thermal burns.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
1R01AR063070-01A1
Application #
8502074
Study Section
Arthritis, Connective Tissue and Skin Study Section (ACTS)
Program Officer
Baker, Carl
Project Start
2013-04-01
Project End
2018-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
1
Fiscal Year
2013
Total Cost
$458,962
Indirect Cost
$127,618
Name
University of Minnesota Twin Cities
Department
Pediatrics
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Webber, Beau R; O'Connor, Kyle T; McElmurry, Ron T et al. (2017) Rapid generation of Col7a1-/- mouse model of recessive dystrophic epidermolysis bullosa and partial rescue via immunosuppressive dermal mesenchymal stem cells. Lab Invest 97:1218-1224
Skvarova Kramarzova, Karolina; Osborn, Mark J; Webber, Beau R et al. (2017) CRISPR/Cas9-Mediated Correction of the FANCD1 Gene in Primary Patient Cells. Int J Mol Sci 18:
Mallhi, K; Orchard, P J; Miller, W P et al. (2017) Non-myeloablative conditioning for second hematopoietic cell transplantation for graft failure in patients with non-malignant disorders: a prospective study and review of the literature. Bone Marrow Transplant 52:726-732
Boyle, William S; Senger, Kyle; Tolar, Jakub et al. (2017) Heparin Enhances Transfection in Concert with a Trehalose-Based Polycation with Challenging Cell Types. Biomacromolecules 18:56-67
Agrawal, Pranav; Ingle, Nilesh P; Boyle, William S et al. (2016) Fast, Efficient, and Gentle Transfection of Human Adherent Cells in Suspension. ACS Appl Mater Interfaces 8:8870-4
Boull, Christina L; Hylwa, Sara A; Sajic, Dusan et al. (2016) Toxic Epidermal Necrolysis in Recessive Dystrophic Epidermolysis Bullosa following Bone Marrow Transplantation. J Pediatr 173:242-4
Osborn, Mark J; Belanto, Joseph J; Tolar, Jakub et al. (2016) Gene editing and its application for hematological diseases. Int J Hematol 104:18-28
Osborn, Mark J; Webber, Beau R; Knipping, Friederike et al. (2016) Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases. Mol Ther 24:570-81
Webber, Beau R; Osborn, Mark J; McElroy, Amber N et al. (2016) CRISPR/Cas9-based genetic correction for recessive dystrophic epidermolysis bullosa. NPJ Regen Med 1:
Tolarová, Markéta; McGrath, John A; Tolar, Jakub (2016) Venturing into the New Science of Nucleases. J Invest Dermatol 136:742-5

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