Advances in molecular genetics have made it possible to correct genetic diseases by introducing a non-defective gene into the afflicted host via somatic cells. The objective of this proposal is to test the hypothesis that skin is the organ most amenable to correcting many genetic disorders via somatic cell gene therapy, especially those characterized by deficiency of a plasma protein. We propose to take advantage of a unique animal model to examine the possibility of curing diseases by use of genetically modified skin. Murine hypotransferrinemia is a recessive disorder resulting from a splicing defect in the plasma transferrin gene; transferrin being the major iron binding protein in plasma. Homozygotes for this disorder have drastically reduced levels of transferrin, but demonstrate iron overload disease of the liver, heart and pancreas. These mutants exhibit severe anemia (a faithful replica of the rare human disease atransferrinemia) and at the same time manifest the complications of the more common disease, hemochromatosis. We propose to evaluate the ability of skin implants, expressing transferrin, to provide the absent plasma protein and thereby alleviate the clinical manifestations of severe hypotransferrinemia. Fibroblasts taken from homozygote mice will be transduced in vitro with a retrovirus plasmid containing the cDNA for either human or murine transferrin. The genetically modified fibroblasts will be implanted subdermally and the concentration of plasma transferrin as well as objective indices of the disease state assayed. In a second project we propose to evaluate the use of skin cells expressing a truncated version transferrin to manage iron overload diseases. A number of diseases exist in which iron overload can not be treated by conventional phlebotomies, thalassemia being the most notable. We hypothesize that a shortened transferrin molecule, a truncated transferrin, would act as a perfect iron chelator. It wound bind iron with high affinity, not be recognized by transferrin receptors, and would be excreted in the urine. We propose to graft a dermal matrix of genetically modified fibroblasts expressing a truncated transferrin gene beneath the skin of homozygote hypotransferrinemic mice and assay these transfected mice for reduction in parenchymal tissue iron. We feel that our previous studies on iron metabolism, the existence of the mutant mouse and the fact that the relevant genes are in hand, offers a unique opportunity to examine the use of skin cells in somatic cell gene therapy.

Project Start
Project End
Budget Start
1995-10-01
Budget End
1996-09-30
Support Year
5
Fiscal Year
1996
Total Cost
Indirect Cost
Name
University of Utah
Department
Type
DUNS #
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Davis, Howard E; Rosinski, Matthew; Morgan, Jeffrey R et al. (2004) Charged polymers modulate retrovirus transduction via membrane charge neutralization and virus aggregation. Biophys J 86:1234-42
Erdag, Gulsun; Medalie, Daniel A; Rakhorst, Hinne et al. (2004) FGF-7 expression enhances the performance of bioengineered skin. Mol Ther 10:76-85
Erdag, Gulsun; Morgan, Jeffrey R (2004) Allogeneic versus xenogeneic immune reaction to bioengineered skin grafts. Cell Transplant 13:701-12
Woodley, David T; Krueger, Gerald G; Jorgensen, Cynthia M et al. (2003) Normal and gene-corrected dystrophic epidermolysis bullosa fibroblasts alone can produce type VII collagen at the basement membrane zone. J Invest Dermatol 121:1021-8
Erdag, Gulsun; Morgan, Jeffrey R (2002) Interleukin-1alpha and interleukin-6 enhance the antibacterial properties of cultured composite keratinocyte grafts. Ann Surg 235:113-24
Hamoen, Karen E; Morgan, Jeffrey R (2002) Transient hyperproliferation of a transgenic human epidermis expressing hepatocyte growth factor. Cell Transplant 11:385-95
Davis, Howard E; Morgan, Jeffrey R; Yarmush, Martin L (2002) Polybrene increases retrovirus gene transfer efficiency by enhancing receptor-independent virus adsorption on target cell membranes. Biophys Chem 97:159-72
DeWitt, Ann; Iida, Tomoko; Lam, Ho-Yan et al. (2002) Affinity regulates spatial range of EGF receptor autocrine ligand binding. Dev Biol 250:305-16
Gill, Pritmohinder S; Krueger, Gerald G; Kohan, Donald E (2002) Doxycycline-inducible retroviral expression of green fluorescent protein in immortalized human keratinocytes. Exp Dermatol 11:266-74
Erdag, Gulsun; Morgan, Jeffrey R (2002) Survival of fetal skin grafts is prolonged on the human peripheral blood lymphocyte reconstituted-severe combined immunodeficient mouse/skin allograft model. Transplantation 73:519-28

Showing the most recent 10 out of 42 publications