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Molecular Dermatology pp 425–440Cite as

Cell- and Protein-Based Therapy Approaches for Epidermolysis Bullosa

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 961))

Abstract

Dystrophic epidermolysis bullosa (DEB) is a clinically heterogeneous heritable skin fragility disorder characterized by mechanically induced mucocutaneous blistering. On the molecular level DEB is caused by mutations leading to deficiency in collagen VII (CVII), a large extracellular protein building anchoring fibrils that attach the epidermis to the dermis. Severely affected patients suffer from wounds, which heal with excessive scarring causing mutilating deformities of hands and feet. The patients are also predisposed to development of aggressive squamous cell carcinomas at sites of chronic wounds. Currently no available therapies exist for this extremely disabling and stigmatizing disorder. We are developing and evaluating cell- and protein-based therapies for the management of DEB.

Dermal fibroblasts are easy to propagate in vitro, they produce CVII, and they have immunomodulating capacities, which makes it possible to use allogeneic fibroblasts for therapy without risking major adverse effects from the host’s immune system. Hence, fibroblasts, and fibroblast-like cells such as mesenchymal stromal cells, are prime candidates for cell-based DEB therapies.

An alternative for management of disorders caused by defects in proteins with relatively low turnover rate is to introduce the protein de novo to the tissue by direct application of the protein. CVII is long-lived and expressed in moderate amounts in the skin; this makes injection of collagen VII protein a realistic approach for the treatment of DEB.

Here we present methods and protocols that we are using for fibroblast- and recombinant CVII-based therapies of DEB in our model of this disease, the CVII hypomorphic mouse. These protocols are directed towards management of DEB but they can be easily adapted for the treatment of other skin fragility disorders.

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Abbreviations

A1AT:

α-1-Antitrypsin

BSA:

Bovine serum albumin

CVII:

Collagen VII

DAPI:

4′,6-Diamidino-2-phenylindole

DEB:

Dystrophic epidermolysis bullosa

DEJ:

Dermal–epidermal junction

DMEM:

Dulbecco’s modified Eagle’s medium

EB:

Epidermolysis bullosa

EDTA:

Ethylenediaminetetraacetic acid

EGFP:

Enhanced green fluorescent protein

FCS:

Fetal calf serum

H&E:

Haematoxylin and eosin

OCT:

Optimal cutting temperature compound

PBS:

Phosphate-buffered saline

PBS-T:

Phosphate-buffered saline Tween-20

PFA:

Paraformaldehyde

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Acknowledgments

This work was supported in part by the Excellence Initiative of the German Federal and State Governments (Freiburg Institute for Advanced Studies, FRIAS, School of Life Sciences), by the “Network Epidermolysis bullosa” grant and the “Stem cell therapy for inherited skin fragility disorders” grant from the Federal Ministry for Education and Research (BMBF) and by Debra International.

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Authors and Affiliations

  1. Department of Dermatology, University Freiburg Medical Center, Freiburg, Germany

    Alexander Nyström & Johannes S. Kern

  2. Freiburg Institute for Advanced Studies, School of Life Sciences, Center for Biological Systems Analysis, University of Freiburg, Freiburg, Germany

    Leena Bruckner-Tuderman

  3. Department of Dermatology, University Medical Center Freiburg, Freiburg, Germany

    Leena Bruckner-Tuderman

Authors
  1. Alexander Nyström
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  2. Leena Bruckner-Tuderman
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  3. Johannes S. Kern
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Corresponding author

Correspondence to Leena Bruckner-Tuderman .

Editor information

Editors and Affiliations

  1. Universitäts-Hautklinik, Department of Molecular Dermatology, Universitätsklinikum Freiburg, Hauptstr. 7, Freiburg, 79104, Germany

    Cristina Has

  2. Universitäts-Hautklinik, Department of Molecular Dermatology, Universitätklinikum Freiburg, Hauptstr. 7, Freiburg, 79104, Germany

    Cassian Sitaru

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Nyström, A., Bruckner-Tuderman, L., Kern, J.S. (2013). Cell- and Protein-Based Therapy Approaches for Epidermolysis Bullosa. In: Has, C., Sitaru, C. (eds) Molecular Dermatology. Methods in Molecular Biology, vol 961. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-227-8_29

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  • DOI: https://doi.org/10.1007/978-1-62703-227-8_29

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-226-1

  • Online ISBN: 978-1-62703-227-8

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