Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2020 May 1;12(5):a035667.
doi: 10.1101/cshperspect.a035667.

Toward Combined Cell and Gene Therapy for Genodermatoses

Laura De Rosa  1 , Maria Carmela Latella  1 , Alessia Secone Seconetti  1 , Cecilia Cattelani  2 , Johann W Bauer  3 , Sergio Bondanza  1 , Michele De Luca  2
Affiliations
Review

Toward Combined Cell and Gene Therapy for Genodermatoses

Laura De Rosa et al. Cold Spring Harb Perspect Biol. .

Abstract

To date, more than 200 monogenic, often devastating, skin diseases have been described. Because of unmet medical needs, development of long-lasting and curative therapies has been consistently attempted, with the aim of correcting the underlying molecular defect. In this review, we will specifically address the few combined cell and gene therapy strategies that made it to the clinics. Based on these studies, what can be envisioned for the future is a patient-oriented strategy, built on the specific features of the individual in need. Most likely, a combination of different strategies, approaches, and advanced therapies will be required to reach the finish line at the end of the long and winding road hampering the achievement of definitive treatments for genodermatoses.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
A view of current ex vivo gene therapy strategies for genodermatoses. Epidermal stem cells, fibroblasts, and induced pluripotent stem cells (iPS) can be genetically modified by means of gene addition (the most frequent approach), gene replacement, and allele-specific gene knockout (KO) for both recessively and dominantly inherited genetic skin diseases.
Figure 2.
Figure 2.
Schematic representation of human skin and epidermolysis bullosa (EB). (A) Epidermal layers (left) and the epidermal–dermal junction (middle) are designated. The right panel denotes the hemidesmosome and its components, in relation to EB simplex (EBS), junctional EB (JEB), and dystrophic EB (DEB). (B) Proteins involved in the pathogenesis of EBS, JEB, and DEB and site of blister formation.
Figure 3.
Figure 3.
Schematic representation of ex vivo gene therapy and clonal analysis. (A) Scheme of combined cell and gene therapy for LAMB3-dependent junctional epidermolysis bullosa (JEB), as reported in Hirsch et al. (2017). Clonogenic keratinocytes, consisting of stem cells (rhodamine red) and transient amplyfing cells (light pink), were cultivated from a skin biopsy, transduced with γRVs, containing LAMB3 and used to prepare transgenic epidermal sheets, which were then transplanted on surgically prepared skin lesions. Of note, clonal tracing performed on cultures initiated from the restored skin (Hirsch et al. 2017) has shown that long-term skin regeneration (right part of the panel) is sustained only by long-lived, self-renewing stem cells detected as holoclone-forming cells (see panel B), as indicated by the rhodamine red color. (B) Clonal analysis. Keratinocytes are inoculated (0.5 cells per well) onto 96-multiwell plates containing irradiated 3T3-J2 cells. After 7 days of cultivation, single clones are transferred to two dishes and cultivated. One dish (one-quarter of the clone) is stained 12 days later for the classification of clonal type, which is determined by the percentage of aborted colonies formed by the progeny of the founding cell. The clone is scored as a holoclone when 0%–5% of colonies are terminal. When 95%–100% of colonies are terminal (or when no colonies formed), the clone is classified as a paraclone. When the number of terminal colonies is between 5% and 95%, the clone is classified as a meroclone. The second dish (three-quarters of the clone) is used for further analyses.
Figure 4.
Figure 4.
Clonal analysis from skin biopsies and primary cultures. Clonal analysis was performed as described in Figure 3B. Human skin samples were obtained as anonymized surgical waste, typically from abdominoplasty or mammoplasty. Skin samples were cut in three parts for (1) direct cloning from intact skin, (2) direct cloning from wounded skin, (3) preparation of primary cultures. (1) Clonal analysis performed directly from the skin biopsy unveiled that the majority of clonogenic cells of intact skin generate holoclones (stem cells). (2) The epidermis was mechanically removed from the center of the biopsy to mimic a wound and the biopsy was kept in the incubator in keratinocyte growth medium (Hirsch et al. 2017). Clonal analysis, performed 24 hours later from a skin section taken in between the two edges of the wound, showed that the majority of clonogenic cells during the wound-healing process generates meroclones and paraclones (transient amplyfing progenitors). A similar distribution of clonogenic cells was detected in primary cultures (3), confirming that such cultures recapitulate a wound-healing process.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Abaci HE, Guo Z, Doucet Y, Jacków J, Christiano A. 2017. Next generation human skin constructs as advanced tools for drug development. Exp Biol Med (Maywood) 242: 1657–1668. 10.1177/1535370217712690 - DOI - PMC - PubMed
    1. Aiuti A, Cattaneo F, Galimberti S, Benninghoff U, Cassani B, Callegaro L, Scaramuzza S, Andolfi G, Mirolo M, Brigida I, et al. 2009. Gene therapy for immunodeficiency due to adenosine deaminase deficiency. N Engl J Med 360: 447–458. 10.1056/NEJMoa0805817 - DOI - PubMed
    1. Aiuti A, Biasco L, Scaramuzza S, Ferrua F, Cicalese MP, Baricordi C, Dionisio F, Calabria A, Giannelli S, Castiello MC, et al. 2013. Lentiviral hematopoietic stem cell gene therapy in patients with Wiskott–Aldrich syndrome. Science 341: 1233151 10.1126/science.1233151 - DOI - PMC - PubMed
    1. Aiuti A, Roncarolo MG, Naldini L. 2017. Gene therapy for ADA-SCID, the first marketing approval of an ex vivo gene therapy in Europe: paving the road for the next generation of advanced therapy medicinal products. EMBO Mol Med 9: 737–740. 10.15252/emmm.201707573 - DOI - PMC - PubMed
    1. Artusi S, Miyagawa Y, Goins WF, Cohen JB, Glorioso JC. 2018. Herpes simplex virus vectors for gene transfer to the central nervous system. Diseases 6: 74 10.3390/diseases6030074 - DOI - PMC - PubMed

Publication types