Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB

Georgiadis, Christos; Syed, Farhatullah; Petrova, Anastasia; Abdul‐Wahab, Alya; Lwin, Su M.; Farzaneh, Farzin; Chan, Lucas; Ghani, Sumera; Fleck, Roland A.; Glover, Leanne; McMillan, James R.; Chen, Mei; Thrasher, Adrian J.; McGrath, John A.; Di, Wei‐Li; Qasim, Waseem

doi:10.1038/jid.2015.364

Cited by 47 publications

(42 citation statements)

References 43 publications

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“…As described in our study for RDEB cells (Fig. 4), and by others (Georgiadis et al, 2016;Goto et al, 2006;Jackow et al, 2016;Titeux et al, 2010), transduced fibroblasts produced COLVII that relocated at the DEJ. However, the long-term persistence of these cells in our TESs remains to be proven.…”

Section: Discussionsupporting

confidence: 86%

Translating the combination of gene therapy and tissue engineering for treating recessive dystrophic epidermolysis bullosa

A¹,

Larouche²,

Ghani³

et al. 2018

eCM

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The combination of gene therapy and tissue engineering is one of the most promising strategies for the treatment of recessive dystrophic epidermolysis bullosa (RDEB). RDEB is a rare genetic disease characterised by mutations in the COL7A1 gene, encoding type VII collagen (COLVII), which forms anchoring fibrils at the dermal-epidermal junction of the skin. This disease causes severe blistering and only palliative treatments are offered. In this study, the base of a strategy combining gene therapy and a tissue-engineered skin substitute (TES), which would be suitable for the permanent closure of skin wounds, was set-up. As a high transduction efficiency into fibroblasts and/or keratinocytes seems to be a prerequisite for a robust and sustained correction of RDEB, different envelope pseudotyped retroviral vectors and the transduction enhancer EF-C were tested. When green fluorescent protein (GFP) was used as a reporter gene to evaluate the retroviral-mediated gene transfer, the fibroblast infection efficiency was 30 % higher with the Ampho pseudotyped vector as compared with the other pseudotypes. At least a 3.1-fold and a 1.3-fold increased transduction were obtained in fibroblasts and keratinocytes, respectively, with EF-C as compared with polybrene. A continuous and intense deposit of haemagglutinin (HA)-COLVII was observed at the dermal-epidermal junction of self-assembled TESs made of cells transduced with a HA-tagged COL7A1 vector. Furthermore, HA-tagged basal epidermal cells expressing keratin 19 were observed in TESs, suggesting stem cell transduction. This approach could be a valuable therapeutic option to further develop, in order to improve the long-term life quality of RDEB patients.

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Section: Discussionsupporting

confidence: 86%

Translating the combination of gene therapy and tissue engineering for treating recessive dystrophic epidermolysis bullosa

A¹,

Larouche²,

Ghani³

et al. 2018

eCM

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“…Here, we used cells that in a differentiated state produce physiological levels of correctly folded collagen VII (Kuhl et al, 2015). Introduction of collagen VII above physiological levels-for example, by using cells genetically modified to overexpress collagen VII or injecting pure recombinant collagen VII (Georgiadis et al, 2016;Hou et al, 2015)-may alter its stability by altering the ratio between collagen VII and its interaction partners or other proteins promoting collagen VII stability. Thus, the pharmacokinetic behavior of collagen VII likely has to be individually determined for each therapeutic approach.…”

Section: Discussionmentioning

confidence: 99%

Collagen VII Half-Life at the Dermal-Epidermal Junction Zone: Implications for Mechanisms and Therapy of Genodermatoses

Kühl

Mezger

Haußer

et al. 2016

Journal of Investigative Dermatology

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The tissue half-life of proteins largely determines treatment frequency of non-gene-editing-based therapies targeting the cause of genodermatoses. Surprisingly, such knowledge is missing for a vast number of proteins involved in pathologies. The dermal-epidermal junction zone is believed to be a rather static structure, but to our knowledge no detailed analysis of the stability of proteins within this zone has been performed. Here, we addressed the in vivo half-life of collagen type VII using genetic ablation of its expression and therapeutic introduction of exogenous collagen VII in a preclinical model. A similar in vivo stability of collagen VII was observed in the skin, tongue, and esophagus, with a half-life of about 1 month. Collagen VII expressed by intradermally injected mesenchymal stromal cells also exhibited a similar half-life. Our study provides key information needed for the development of protein replacement or cell-based therapies for dystrophic epidermolysis bullosa caused by genetic deficiency of collagen VII. Moreover, by showing what we define as an intermediate half-life of collagen VII, our study challenges the view of the dermal-epidermal junction zone as a static structure with very slow turnover.

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“…In previous studies, we and others have shown the capability of genetically corrected fibroblasts to improve skin cohesion in RDEB skin. Genetic modification was previously performed using a lentiviral vector (Chen et al, 2002;Georgiadis et al, 2016), a bacteriophage integrase (Ortiz-Urda et al, 2003b;Ortiz-Urda et al, 2002), and gamma retroviruses (Goto et al, 2006;Titeux et al, 2010). In these works, gene-corrected RDEB fibroblasts were delivered either by local injections (Ortiz-Urda et al, 2003;Woodley et al, 2003), intravenous injections (Woodley et al, 2007), or through transplanted skin equivalent (SE) using genetically corrected fibroblasts and keratinocytes (Goto et al, 2006;Titeux et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector

Jacków

Titeux

Portier

et al. 2016

Journal of Investigative Dermatology

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Patients with recessive dystrophic epidermolysis bullosa (RDEB) lack type VII collagen and therefore have severely impaired dermal-epidermal stability causing recurrent skin and mucosal blistering. There is currently no specific approved treatment for RDEB. We present preclinical data showing that intradermal injections of genetically corrected patient-derived RDEB fibroblasts using a Good Manufacturing Practices grade self-inactivating COL7A1 retroviral vector reverse the disease phenotype in a xenograft model in nude mice. We obtained 50% transduction efficiency in primary human RDEB fibroblasts with an average low copy number (range = 1-2) of integrated provirus. Transduced fibroblasts showed strong type VII collagen re-expression, improved adhesion properties, normal proliferative capabilities, and viability in vitro. We show that a single intradermal injection of 3 × 10(6) genetically corrected RDEB fibroblasts beneath RDEB skin equivalents grafted onto mice allows type VII collagen deposition, anchoring fibril formation at the dermal-epidermal junction, and improved dermal-epidermal adherence 2 months after treatment, supporting functional correction in vivo. Gene-corrected fibroblasts previously showed no tumorigenicity. These data show the efficacy and safety of gene-corrected fibroblast therapy using a self-inactivating vector that has now been good manufacturing grade-certified and pave the way for clinical translation to treat nonhealing wounds in RDEB patients.

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Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB

Cited by 47 publications

References 43 publications

Translating the combination of gene therapy and tissue engineering for treating recessive dystrophic epidermolysis bullosa

Translating the combination of gene therapy and tissue engineering for treating recessive dystrophic epidermolysis bullosa

Collagen VII Half-Life at the Dermal-Epidermal Junction Zone: Implications for Mechanisms and Therapy of Genodermatoses

Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector

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