Treatment of Posterior Hypospadias by the Autologous Graft of Cultured Urethral Epithelium

Romagnoli, Giuseppe; Luca, Michele De; Faranda, F; Bandelloni, Roberto; Franzi, Adriano T.; Cataliotti, F; Cancedda, Ranieri

doi:10.1056/nejm199008233230806

Cited by 107 publications

(35 citation statements)

References 13 publications

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“…Thus, autologous epithelia made ex vivo are now used to treat autologous burn patients and chronic ulcers (Romagnoli et al, 1990). This technology provides the possibility of exploring innovative therapeutic approaches for inherited and acquired diseases by the use of implants of genetically modified keratinocytes that permanently express biologically active proteins (Mathor et al, 1996;Kolodka et al, 1998).…”

Section: Introductionmentioning

confidence: 97%

Construction of Skin Equivalents for Gene Therapy of Recessive Dystrophic Epidermolysis Bullosa

et al. 2004

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We have shown that retroviral vectors efficiently transfer the 9-kb collagen type VII cDNA into keratinocytes of dogs with recessive dystrophic epidermolysis bullosa (RDEB) and achieve correction of the RDEB phenotype in vitro. As a next step toward gene therapy applications, we have assessed the suitability of retroviral vectors to transduce human collagen type VII cDNA into primary human RDEB keratinocytes and generate transplantable autologous skin equivalents. The transduced RDEB keratinocytes permanently express high levels of recombinant collagen type VII that assembles into functional homotrimers readily secreted into the extracellular matrix. The recombinant collagen type VII reverts the migration and invasion potential of the transduced RDEB keratinocytes in vitro and is efficiently deposited at the dermal epidermal junction of artificial skin prepared with the reverted cells and artificial dermis made of biomaterial sponges embedded with dermal RDEB fibroblasts. Transplantable fibrin-based skin equivalents made with the transduced RDEB keratinocytes and grafted onto SCID mice either orthotopically or in accordance with the flap method generated cohesive and orderly stratified epithelia with all the characteristics of normal human epidermis, including rapid formation of anchoring fibrils. Because transplantable epithelia are routinely used to cure patients suffering from large skin or mucosal defects, the full phenotypic reversion of primary RDEB epidermal clonogenic cells mediated by recombinant retroviral vectors opens new perspectives in the long-term treatment of genodermatoses.

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Section: Introductionmentioning

confidence: 97%

Construction of Skin Equivalents for Gene Therapy of Recessive Dystrophic Epidermolysis Bullosa

et al. 2004

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“…Cultured epithelial sheets are routinely used to make autologous grafts for patients suffering from large skin and mucosal defects; the engrafted stem cells assure the persistence of the regenerated epidermis during the patient's lifetime (7)(8)(9). Epidermal keratinocytes synthesize and secrete several gene products that might accomplish autocrine, paracrine, and even endocrine functions (10).…”

mentioning

confidence: 99%

Clonal analysis of stably transduced human epidermal stem cells in culture.

Mathor

Ferrari

Dellambra

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

156

147

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We have transduced normal human keratinocytes with retroviral constructs expressing a bacterial .8-galactosidase (,B-gal) gene or a human cDNA under control of a long terminal repeat. Efficiency of gene transfer averaged approximately 50%Yo and 95% of clonogenic keratinocytes for ,B-gal and hIL-6, respectively. Both genes were stably integrated and expressed for more than 150 generations. Clonal analysis showed that both holoclones and their transient amplifying progeny expressed the transgene permanently. Southern blot analysis on isolated clones showed that many keratinocyte stem cells integrated multiple proviral copies in their genome and that the synthesis of the exogenous gene product in vitro was proportional to the number of proviral integrations. When cohesive epidermal sheets prepared from stem cells transduced with hIL-6 were grafted on athymic animals, the serum levels of hIL-6 were strictly proportional to the rate of secretion in vitro and therefore to the number of proviral integrations. The possibility of specifying the level of transgene expression and its permanence in a homogeneous clone of stem cell origin opens new perspectives in the long-term treatment of genetic disorders.Cultured human keratinocytes generate cohesive sheets of epithelium (1), which maintain the characteristics of the original donor site (2, 3) and retain stem cells, namely cells that upon division replace their own number and also give rise to cells that differentiate further into one or more specialized types (4-6). Cultured epithelial sheets are routinely used to make autologous grafts for patients suffering from large skin and mucosal defects; the engrafted stem cells assure the persistence of the regenerated epidermis during the patient's lifetime (7-9). Epidermal keratinocytes synthesize and secrete several gene products that might accomplish autocrine, paracrine, and even endocrine functions (10). Indeed, it has been unambiguously demonstrated that keratinocyte-derived polypeptides cross the basement membrane and are secreted into the bloodstream (11, 12). Basal keratinocytes are also sources of adhesion molecules and matrix proteins, which form the anchoring structures of the epidermis, and some of these proteins are affected in genetic skin diseases (13). Therefore, epidermal cells are very attractive targets for the gene therapy of skin diseases, as well as for systemic delivery of recombinant proteins for the treatment of a number of genetic disorders.Ex vivo transduced mammalian epidermal cells, including human keratinocytes, have been successfully generated (14-16). Protein production in vitro and/or secretion into the circulation by keratinocytes grafted onto animals has been shown for exogenous growth hormone (17-20), neomycin phosphotransferase (21), steroid sulfatase (22), factor IX (23), al-anti-trypsin (24), 13-chorionic gonadotropin (15), and ,B-galactosidase (13-gal) (19,(24)(25)(26) after transduction with replication-deficient retroviral or adenoviral vectors. However, none of these approa...

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“…Epithelial cells, isolated from a biopsy taken from the urethral meatus, reconstitute in vitro an urethral stratified squamous epithelium that offers an alternative in the urethral reconstruction of patients suffering from posterior hypospadias. 144,145 Moreover, these cells have been shown to retain the characteristics of a normal urothelium. 144,145 Nevertheless, the main limitation of this procedure is the difficulty in implanting such a delicate epithelium.…”

Section: Urotheliummentioning

confidence: 98%

“…144,145 Moreover, these cells have been shown to retain the characteristics of a normal urothelium. 144,145 Nevertheless, the main limitation of this procedure is the difficulty in implanting such a delicate epithelium. Devices such as tubular polytetrafluoroethylene supports 145 and silicone catheters, 146 which may be implanted subcutaneously, have been investigated to improve the feasibility of urethra reconstruction.…”

Section: Urotheliummentioning

confidence: 98%

Tissue Engineering for Damaged Surface and Lining Epithelia: Stem Cells, Current Clinical Applications, and Available Engineered Tissues

Guerra

Dellambra

Panacchia

et al. 2009

Tissue Engineering Part B: Reviews

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Tissue engineering is an important tool for the treatment of damaged surface and lining epithelia. A source of cells and biocompatible substrates upon which cells can grow and differentiate are key components of this technology. Cultured normal human epithelial cells reconstitute sheets of stratified epithelia that retain biochemical and histological characteristics as well as specific differentiation features of the original donor site. Maintenance of epithelial stem cells in culture and a well-prepared receiving wound bed allow to permanently regenerate full-thickness wounds by means of in vitro reconstituted epithelia. Further, cultured cells produce growth factors and extracellular matrix (ECM) components that help resident cells to contribute to the wound-healing process. Biological matrices enhance the performance of the in vitro reconstituted epithelia. Owing to their similarity to the ECM, natural polymers offer the advantage of being similar to macromolecules that the human environment is prepared to recognize. They also maintain biological information and physical and chemical features that are instructive for cells used to populate them. This article discusses the developments of tissues engineered for cutaneous and mucosal regeneration. Native tissues and their stem cells are also considered, to enhance understanding of the extensive field of tissue reconstruction.

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Treatment of Posterior Hypospadias by the Autologous Graft of Cultured Urethral Epithelium

Cited by 107 publications

References 13 publications

Construction of Skin Equivalents for Gene Therapy of Recessive Dystrophic Epidermolysis Bullosa

Construction of Skin Equivalents for Gene Therapy of Recessive Dystrophic Epidermolysis Bullosa

Clonal analysis of stably transduced human epidermal stem cells in culture.

Tissue Engineering for Damaged Surface and Lining Epithelia: Stem Cells, Current Clinical Applications, and Available Engineered Tissues

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