Induced Pluripotent Stem Cells from Individuals with Recessive Dystrophic Epidermolysis Bullosa

Division of Hematology-Oncology, Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Journal of Investigative Dermatology (Impact Factor: 7.22). 12/2010; 131(4):848-56. DOI: 10.1038/jid.2010.346
Source: PubMed


Recessive dystrophic epidermolysis bullosa (RDEB) is an inherited blistering skin disorder caused by mutations in the COL7A1 gene-encoding type VII collagen (Col7), the major component of anchoring fibrils at the dermal-epidermal junction. Individuals with RDEB develop painful blisters and mucosal erosions, and currently, there are no effective forms of therapy. Nevertheless, some advances in patient therapy are being made, and cell-based therapies with mesenchymal and hematopoietic cells have shown promise in early clinical trials. To establish a foundation for personalized, gene-corrected, patient-specific cell transfer, we generated induced pluripotent stem (iPS) cells from three subjects with RDEB (RDEB iPS cells). We found that Col7 was not required for stem cell renewal and that RDEB iPS cells could be differentiated into both hematopoietic and nonhematopoietic lineages. The specific epigenetic profile associated with de-differentiation of RDEB fibroblasts and keratinocytes into RDEB iPS cells was similar to that observed in wild-type (WT) iPS cells. Importantly, human WT and RDEB iPS cells differentiated in vivo into structures resembling the skin. Gene-corrected RDEB iPS cells expressed Col7. These data identify the potential of RDEB iPS cells to generate autologous hematopoietic grafts and skin cells with the inherent capacity to treat skin and mucosal erosions that typify this genodermatosis.

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Article: Induced Pluripotent Stem Cells from Individuals with Recessive Dystrophic Epidermolysis Bullosa

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    • "To date, iPSCs for various genetic diseases have been developed, such as for certain type of Parkinson's disease [5], spinal muscular atrophy [6], lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary valve stenosis, abnormal genitalia, retardation of growth, and deafness (LEOPARD) syndrome [7], long Q-T syndrome [8], Timothy syndrome [9], Hurler syndrome [10], epidermolysis bullosa [11], and thalassemia [12]. "
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    ABSTRACT: In genetic diseases, where the cells are already damaged, the damaged cells can be replaced by new normal cells, which can be differentiated from iPSC. To avoid immune rejection, iPSC from the patient's own cell can be developed. However, iPSC from the patients's cell harbors the same genetic aberration. Therefore, before differentiating the iPSCs into required cells, genetic repair should be done. This review discusses the various technologies to repair the genetic aberration in patient-derived iPSC, or to prevent the genetic aberration to cause further damage in the iPSC-derived cells, such as Zn finger and TALE nuclease genetic editing, RNA interference technology, exon skipping, and gene transfer method. In addition, the challenges in using the iPSC and the strategies to manage the hurdles are addressed.
    02/2012; 2012:498197. DOI:10.1155/2012/498197
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    • "Keratinocyte-derived human iPS cells could be differentiated into pancreatic endoderm with an efficiency that was comparable to that for human embryonic stem cells (Santamaria et al, 2010). Recent reports demonstrate that iPS cells can differentiate into keratinocytes (Bilousova et al, 2011; Tolar et al, 2011). The advantages of iPS cells over embryonic stem cells for regeneration therapy include the lack of ethical issues and, since iPS cells can be autologous, the elimination of immune rejection concerns. "
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    ABSTRACT: This is a chronicle of concepts in the field of epidermal stem cell biology and a historic look at their development over time. The past 25 years have seen the evolution of epidermal stem cell science, from first fundamental studies to a sophisticated science. The study of epithelial stem cell biology was aided by the ability to visualize the distribution of stem cells and their progeny through lineage analysis studies. The excellent progress we have made in understanding epidermal stem cell biology is discussed in this article. The challenges we still face in understanding epidermal stem cells include defining molecular markers for stem and progenitor sub-populations, determining the locations and contributions of the different stem cell niches, and mapping regulatory pathways of epidermal stem cell proliferation and differentiation. However, our rapidly evolving understanding of epidermal stem cells has many potential uses that promise to translate into improved patient therapy.
    Journal of Investigative Dermatology 12/2011; 132(3 Pt 2):797-810. DOI:10.1038/jid.2011.434 · 7.22 Impact Factor
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    • "The obtaining of differentiated cells from iPS or hES is still a major challenge. Recently, iPS cells have been obtained from EB patients (Tolar et al., 2011) and it is expected that iPSs from other skin diseases will soon be generated. So far, iPS cells differentiation to fully functional keratinocytes, as performed with hES, has not been reported. "
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    ABSTRACT: Over the past decades, skin has become an increasingly interesting target for replacement therapies. Easy access plays a pivotal role in its widespread use in this context. Current cell culture techniques have optimized in vitro expansion of cells obtained from skin biopsies to be assembled in three-dimensional matrices and engineered skin equivalents that are amenable to clinical use. A wide range of natural scaffolds and synthetic materials are now available as matrices in organotypic skin cultures for skin regeneration (Shevchenko et al., 2010). Patients with severe skin loss require large-scale production of composite skin equivalents. We developed an improved whole autologous bioengineered skin based on the use of a fibrin three-dimensional dermal scaffold in which fibroblasts are embedded (World Patent WO/2002/072800) (Figure 1). We provided evidence that this plasma-based dermal equivalent adequately supports keratinocyte growth (Meana et al., 1998). Immunohistochemical studies over long follow-up periods showed that experimental grafting on immunodeficient mice yielded a healthy and mature skin with human architecture that persisted even after several epidermal turn-overs (Llames et al., 2004). Permanent skin regeneration requires preservation of epidermal cell stemness. The preclinical model fulfils this requirement (Larcher et al., 2007). Bioengineered human skin has been successfully employed in a clinical scenario (Figure 1) for permanent coverage in the case of extensive burns, necrotizing fascitis, removal of giant nevi, and graft-versus-host disease (Llames et al., 2004; 2006; Gómez et al., 2011). Currently, the use of bioengineered skin has spread to a wider range of applications such as the management of injuries of different aetiology including vascular and diabetic wounds and more recently the treatment of wounds associated with genetic rare diseases such as epidermolysis bullosa (EB). EB is characterized by skin blistering following minor friction or mechanical trauma. The condition varies from limited blisters in the skin to a form involving internal epithelial lining. The management of EB is mainly supportive with symptomatic treatment, since currently no cure exists. Nevertheless, EB patients may benefit from the treatment with new cell-based therapies. In this context, the EMEA awarded the Orphan Drug Designation to a chimerical version of the substitute (orphan designation number EU/306/369). Two additional strategies to treat EB, based on the use of bioengineered skin, are being explored by our team. Our approach to study the physiopathology of the skin evolved also toward disease modeling. We have established a skin-humanized mouse model system based on bioengineered human skin-engrafted immunodeficient mice (Del Rio et al., 2002b; Llames et al., 2004). This chimerical model involves the regeneration of human skin, vascularized and innervated by mouse vessels and nerves. This method allows for the generation of a large number of engrafted mice containing a significant area of homogeneous single donor-derived human skin in a relatively short period of time. We have deconstructed-reconstructed skin disorders using skin cells isolated from healthy donor or patient biopsies. Our work included different rare human monogenic skin diseases, such as the recessive form of dystrophic Epidermolysis Bullosa (RDEB), an inherited mechano-bullous disease (Gache et al., 2004; Spirito et al., 2006), the UV-sensitive cancer-prone disease Xeroderma Pigmentosum (XP) (Garcia et al., 2010), Pachyonychia Congenita (PC) (Garcia et al., 2011) and the Netherton Syndrome (NS) (Di et al., 2011), both debilitating skin disorders. With this model, we have succeeded in reverting the phenotype employing different gene therapy approaches for ex vivo correction of cells. We were also able to generate a skin humanized mouse model of acquired conditions such as psoriasis, a common chronic inflammatory disease where the immune component plays a pivotal role (Guerrero-Aspizua et al., 2010). Finally, the model also serves to conduct studies in normal human skin, both in a physiological or pathological context, to gain insight into a process such as wound healing (Escámez et al., 2004, 2008; Martinez-Santamaría et al., 2009 and unpublished results). These wound healing models also allowed the validation of gene and cell therapy approaches to improve impaired wound healing conditions and to favour the efficacy of bioengineered skin substitutes in tissue regeneration.
    Skin Biopsy - Perspectives, Edited by Uday Khopkar, 11/2011: chapter 14: pages openacces; InTech., ISBN: 978-953-307-290-6
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