Sustainable correction of junctional epidermolysis bullosa via transposon-mediated nonviral gene transfer

Ortiz‐Urda, Susana; Lin, Qun; Yant, Stephen R.; Keene, Douglas R.; Kay, Mark A.; Khavari, Paul A.

doi:10.1038/sj.gt.3301978

Cited by 114 publications

(52 citation statements)

References 29 publications

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“…[21][22][23][24][25][26] However, only limited success with sustainable gene expression has been observed over wide areas of the skin. 11,[27][28][29] Although the mechanism of nucleic acid uptake in keratinocytes is poorly understood, it has been shown in other organ systems that high pressure facilitates delivery. For example, high pressure hydrodynamic intravascular injection produces moderate levels of gene expression in limb muscles.…”

Section: Introductionmentioning

confidence: 99%

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

González-González

Spitler

et al. 2010

Gene Ther

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Nucleic acid-based therapies hold great promise for treatment of skin disorders if delivery challenges can be overcome. To investigate one mechanism of nucleic acid delivery to keratinocytes, a fixed mass of expression plasmid was intradermally injected into mouse footpads in different volumes, and reporter expression was monitored by intravital imaging or skin sectioning. Reporter gene expression increased with higher delivery volumes, suggesting that pressure drives nucleic acid uptake into cells after intradermal injections similar to previously published studies for muscle and liver. For spatiotemporal analysis of reporter gene expression, a dual-axis confocal (DAC) fluorescence microscope was used for intravital imaging following intradermal injections. Individual keratinocytes expressing hMGFP were readily visualized in vivo and initially appeared to preferentially express in the stratum granulosum and subsequently migrate to the stratum corneum over time. Fluorescence microscopy of frozen skin sections confirmed the patterns observed by intravital imaging. Intravital imaging with the DAC microscope is a noninvasive method for probing spatiotemporal control of gene expression and should facilitate development and testing of new nucleic acid delivery technologies.

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Section: Introductionmentioning

confidence: 99%

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

González-González

Spitler

et al. 2010

Gene Ther

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“…This technology has been successfully used to integrate the LAMB3 cDNA into epidermal holoclones from unrelated junctional EB patients. 10 Epidermal holoclones have the greatest reproductive capacity and represent o5% of primary keratinocyte suspensions; they are considered to contain true epidermal stem cells. Non-viral integration of the LAMB3 gene led to normalized levels of laminin-5 protein, hemidesmosome formation and a normal phenotype.…”

Section: Skin Gene Therapy Ur Henggementioning

confidence: 99%

“…Non-viral integration of the LAMB3 gene led to normalized levels of laminin-5 protein, hemidesmosome formation and a normal phenotype. 10 Alternatively, the FC31 bacterial phage integrase, which inserts -albeit with low efficiency-, large genes into genomes was used to stably integrated the type-VII collagen gene into primary epidermal progenitor cells of Figure 3 Summary of genodermatoses. Shown are the epidermis, the basement membrane and the papillary dermis.…”

Section: Skin Gene Therapy Ur Henggementioning

confidence: 99%

Gene therapy progress and prospects: the skin – easily accessible, but still far away

Hengge

2006

Gene Ther

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Significant progress has been made in corrective gene therapy of inherited skin diseases. This includes advances in vector technology, targeted gene expression, gene replacement, and the availability of appropriate animal models for a variety of candidate diseases. In addition, an increased understanding of the uptake and trafficking mechanisms inside keratinocytes has evolved. Topical application facilitates DNA vaccination through the skin, albeit clinical benefits have not yet materialized. However, the translation into clinical trials has only been partially mastered. The latter and the control of immune responses represent challenges for the research community.

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“…Alternatively, fibroblasts (FB) can be used for systemic gene delivery using an ex vivo approach. Genetically modified FB have been used for the systemic delivery of collagen type VII (Goto et al, 2006;Ortiz-Urda et al, 2003;Woodley et al, 2007), transferrin (Petersen et al, 1995), β-glucuronidase (Moullier et al, 1993), and alpha-galactosidase A (Medin et al, 1996). Although these studies have shown promising results and have demonstrated the feasibility of using skin for systemic delivery of protein products, they have not achieved long-term expression of the therapeutic protein.…”

Section: Introductionmentioning

confidence: 99%

“…Since the genetic basis of many inheritable skin diseases is known, a number of preclinical gene therapy studies have been conducted to treat acquired and inherited skin diseases. In cell culture and animal models (immunocompromised mice and dogs), efficient gene transfer of the corrective gene into skin cells has been used to treat junctional epidermolysis bullosa (JEB; Dellambra et al, 1998;Seitz et al, 1999;Spirito et al, 2006;Vailly et al, 1998), dystrophic epidermolysis bullosa (Baldeschi et al, 2003;Chen et al, 2002;Chen et al, 2000;Ortiz-Urda et al, 2003;Ortiz-Urda et al, 2002), xeroderma pigmentosum (Arnaudeau-Begard et al, 2003), X-linked ichtyosis (Freiberg et al, 1997;Jensen et al, 1993), as well as lamellar ichthyosis (Choate et al, 1996). In a recent clinical trial, a normal laminin 5-β3 gene was introduced into KC from a JEB patient using a retroviral vector, and the genetically modified KC was subsequently grafted back onto the patient.…”

mentioning

confidence: 99%

An Approach to Achieve Long-Term Expression in Skin Gene Therapy

2008

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For gene therapy purposes, the skin is an attractive organ to target for systemic delivery of therapeutic proteins to treat systemic diseases, skin diseases, or skin cancer. To achieve long-term stable expression of a therapeutic gene in keratinocytes (KC), we have developed an approach using a bicistronic retroviral vector expressing the desired therapeutic gene linked to a selectable marker (multidrug resistant gene, MDR) that is then introduced into KC and fibroblasts (FB) to create genetically modified human skin equivalent (HSE). After grafting the HSE onto immunocompromised mice, topical colchicine treatment is used to select and enrich for genetically modified keratinocyte stem cells (KSC) that express MDR and are resistant to colchicine's antimitotic effects. Both the apparatus for topical colchicine delivery and the colchicine doses have been optimized for application to human skin. This approach can be validated by systemic delivery of therapeutic factors such as erythropoietin and the antihypertensive atrial natriuretic peptide.

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Sustainable correction of junctional epidermolysis bullosa via transposon-mediated nonviral gene transfer

Cited by 114 publications

References 29 publications

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

Gene therapy progress and prospects: the skin – easily accessible, but still far away

An Approach to Achieve Long-Term Expression in Skin Gene Therapy

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