Michele De Luca

Università degli Studi di Modena e Reggio Emilia, Modène, Emilia-Romagna, Italy

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Publications (115)891.11 Total impact

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    ABSTRACT: Urethral reconstruction has received much attention in recent years, due to pathologies such as recurrence of urethral strictures after treatments. Various surgical techniques have been developed to obtain the best risk-benefit ratio, such as autologous grafts taken from the oral cavity. Tissue engineering and stem cells, growing tissue from a small biopsies, can further improve surgery, reducing invasiveness and morbidity. To determine whether urethra or other epithelia can be equally useful for urethra engineering, a comparison of clonogenic ability, proliferative potential and stem cell markers should be obtained. In this study, 19 biopsies from urethra, and 21 from oral mucosa were obtained from patients, during reconstructive surgery. Urethral and oral tissues were removed from the same donor, to develop primary cultures and cell characterization. The long term regenerative properties of both tissues was investigated in vitro by life span, clonal analysis and markers of different clonal types. Results revealed the same high proliferative potential for urethra and oral mucosa cultures, but maintenance of specific markers. Karyotype and growth factor dependence confirmed the normal phenotype of cultured cells. Clonal analysis of the proliferative compartment highlighted a very different proportion of stem and transient amplifying cells, characterised by dissimilar cell size profile and marker expression. In conclusion, both tissues can be cultured and preserve their stem cells in vitro. Few differences appeared in oral mucosa vs urethra, suggesting that they can be equally useful for tissue engineering of the urethral tract.
    Current Stem Cell Research & Therapy 09/2015; · 2.21 Impact Factor
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    ABSTRACT: In the past few years, substantial preclinical and experimental advances have been made in the treatment of the severe monogenic skin blistering disease epidermolysis bullosa (EB). Promising approaches have been developed in the fields of protein and cell therapies, including allogeneic stem cell transplantation; in addition, the application of gene therapy approaches has become reality. The first ex vivo gene therapy for a junctional EB (JEB) patient was performed in Italy more than 8 years ago and was shown to be effective. We have now continued this approach for an Austrian JEB patient. Further, clinical trials for a gene therapy treatment of recessive dystrophic EB are currently under way in the United States and in Europe. In this review, we aim to point out that sustainable correction of autologous keratinocytes by stable genomic integration of a therapeutic gene represents a realistic option for patients with EB.
    The Keio Journal of Medicine 06/2015; 64(2). DOI:10.2302/kjm.2014-0013-RE
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    ABSTRACT: Background Epidermolysis Bullosa (EB) is caused by mutations in genes that encode proteins belonging to the epidermal-dermal junction assembly. Due to the extreme clinical/genetic heterogeneity of the disease, the current methods available for diagnosing EB involve immunohistochemistry of bioptic samples and transmission electron microscopy followed by single candidate gene Sanger Sequencing (SS), which are labour intensive and expensive clinical pathways.Objectives According to the recently published recommendations for the EB diagnosis and treatment, the assessment of the mutational landscape is now a fundamental step for developing a comprehensive diagnostic path. Next-Generation Sequencing (NGS) via the parallel ultra-deep sequencing of many genes represents a proper method for reducing the processing time and costs of EB diagnostics.Methods We developed an EB disease-comprehensive AmpliSeq panel to accomplish the NGS on the Ion Torrent PGM platform. The panel was performed on ten patients with known genetic diagnoses and was then employed in eight family trios with unknown molecular footprints.ResultsThe panel was successful in finding the causative mutations in all ten of the patients with known mutations, fully confirming the SS data and providing proof of concept of the sensitivity, specificity, and accuracy of this procedure. In addition to being consistent with the clinical diagnosis, it was also effective in the trios, identifying all of the variants, including ones that the SS missed or de novo mutations.Conclusions The NGS and AmpliSeq were shown to be an effective approach for the diagnosis of EB, resulting in a cost- and time-effective 72-hour procedure.This article is protected by copyright. All rights reserved.
    British Journal of Dermatology 04/2015; 173(3). DOI:10.1111/bjd.13858 · 4.28 Impact Factor
  • Graziella Pellegrini · Michele De Luca ·
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    ABSTRACT: Corneal diseases and blindness can result from deficiency in limbal stem cells, and autologous transplantation is often the only therapeutic option. Two recent studies in Nature have provided insights into regulation of limbal stem cell function and corneal regeneration and present new opportunities for clinical interventions in eye diseases.
    Cell Stem Cell 08/2014; 15(2):121-122. DOI:10.1016/j.stem.2014.07.011 · 22.27 Impact Factor
  • Paolo Bianco · Elena Cattaneo · Michele De Luca · Luca Pani ·

    Nature 02/2014; 506(7489):434. DOI:10.1038/506434c · 41.46 Impact Factor
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    ABSTRACT: We report a long-term follow-up (6.5 years) of a phase I/II clinical trial envisaging the use of autologous genetically modified cultured epidermal stem cells for gene therapy of junctional epidermolysis bullosa, a devastating genetic skin disease. The critical goals of the trial were to evaluate the safety and long-term persistence of genetically modified epidermis. A normal epidermal-dermal junction was restored and the regenerated transgenic epidermis was found to be fully functional and virtually indistinguishable from a normal control. The epidermis was sustained by a discrete number of long-lasting, self-renewing transgenic epidermal stem cells that maintained the memory of the donor site, whereas the vast majority of transduced transit-amplifying progenitors were lost within the first few months after grafting. These data pave the way for the safe use of epidermal stem cells in combined cell and gene therapy for genetic skin diseases.
    Stem Cell Reports 01/2014; 2(1):1-8. DOI:10.1016/j.stemcr.2013.11.001 · 5.37 Impact Factor
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    ABSTRACT: Recent breakthroughs in regenerative medicine have generated enthusiasm and many efforts to explore new therapeutic potentials of both somatic and pluripotent stem cells. About 30 years passed since a discovery of a method of producing a great number of human epidermal keratinocytes by cultivation from a small skin biopsy, many possibilities are now envisaged for therapeutic application of different cultured cell types. The importance of stem cell content was proven for many tissues or organs in different pathologies. Ocular burns cause depletion of limbal stem cells, which lead to corneal opacification and visual loss. Most of available treatments are palliative and focused on the relief of the devastating clinical picture. This review is focused on recent developments cell based therapy of limbal stem cell deficiency. All findings can provide support for improvement and standardization of the cure for this disabling disease. Stem Cells 2013.
    Stem Cells 01/2014; 32(1). DOI:10.1002/stem.1517 · 6.52 Impact Factor
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    ABSTRACT: Inherited epidermolysis bullosa (EB) is a family of rare genetic skin disorders characterized by structural and mechanical fragility of skin and mucosal membranes. The main feature of EB is the presence of recurrent skin blistering or erosions, which have a profound impact in the quality of life of EB patients and, in the most severe forms, cause early lethality. During the past two decades, it became possible to identify mutations in genes responsible for different types of EB and characterize the abnormalities of the related proteins. Nowadays, there is no cure for EB; all the treatments are palliative and focused on the relief of the devastating EB clinical picture. Recent advancements in molecular biology, stem cell biology and regenerative medicine have fostered new therapeutic approaches for EB. This review is focused on recent developments in gene therapy, protein replacement and cell-based therapy for EB, all aimed at finding a cure for this devastating disease.
    Regenerative Medicine 07/2013; 8(4):467-81. DOI:10.2217/rme.13.33 · 2.79 Impact Factor
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    ABSTRACT: Aim: Limbal cultures restore the corneal epithelium in patients with ocular burns. We investigated the biological parameters instrumental for their clinical success. Methods: We report a long-term multicenter prospective study on 152 patients carrying corneal destruction due to severe ocular burns, treated with autologous limbal cells cultured on fibrin and clinical-grade 3T3-J2 feeder cells. Clinical results were statistically evaluated both by parametric and nonparametric methods. Results: Clinical outcomes were scored as full success, partial success and failure in 66.05, 19.14 and 14.81% of eyes, respectively. The total number of clonogenic cells, colony size, growth rate and presence of conjunctival cells could not predict clinical results. Instead, the clinical data provided conclusive evidence that graft quality and likelihood of a successful outcome rely on an accurate evaluation of the number of stem cells detected before transplantation as holoclones expressing high levels of the p63 transcription factor. No adverse effects related to the feeder layer have been observed and the regenerated epithelium was completely devoid of any 3T3-J2 contamination. Conclusion: Cultures of limbal stem cells can be safely used to successfully treat massive destruction of the human cornea. We emphasize the importance of a discipline for defining the suitability and the quality of cultured epithelial grafts, which are relevant to the future clinical use of any cultured cell type.
    Regenerative Medicine 06/2013; 8(5). DOI:10.2217/rme.13.43 · 2.79 Impact Factor
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    ABSTRACT: At the time of writing, the Italian Parliament is debating a new law that would make it legal to practice an unproven stem cell treatment in public hospitals. The treatment, offered by a private non-medical organization, may not be safe, lacks a rationale, and violates current national laws and European regulations. This case raises multiple concerns, most prominently the urgent need to protect patients who are severely ill, exposed to significant risks, and vulnerable to exploitation. The scientific community must consider the context-social, financial, medical, legal-in which stem cell science is currently situated and the need for stringent regulation. Additional concerns are emerging. These emanate from the novel climate, created within science itself, and stem cell science in particular, by the currently prevailing model of 'translational medicine'. Only rigorous science and rigorous regulation can ensure translation of science into effective therapies rather than into ineffective market products, and mark, at the same time, the sharp distinction between the striving for new therapies and the deceit of patients.
    The EMBO Journal 05/2013; 32(11). DOI:10.1038/emboj.2013.114 · 10.43 Impact Factor
  • Laura De Rosa · Michele De Luca ·
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    ABSTRACT: It has been unclear whether a uniform group of stem cells gives rise to most cells in the epidermis. A study reveals the presence of at least two stem-cell populations that have different proliferative abilities. See Article p.257
    Nature 09/2012; 489(7415):215-7. DOI:10.1038/489215a · 41.46 Impact Factor
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    ABSTRACT: Cell therapy is an emerging therapeutic strategy aimed at replacing or repairing severely damaged tissues with cultured cells. Specifically, ocular burns cause depletion of limbal stem cells, which leads to corneal opacification and visual loss. Corneal stem cells are segregated in the basal layer of the limbus, which is the transitional zone of the epithelium located between the cornea and the bulbar conjunctiva. Autologous cultured limbal epithelial cells can restore damaged corneas. We sought to establish a culture system that allows preservation of limbal stem cells and preparation of manageable epithelial sheets. We outline some quality criteria, which assure the clinical performance of keratinocyte culture: evaluation of the number of holoclones within a cultured epithelial graft, proportion of aborting colonies, and percentage of cells expressing high levels of ΔNp63α.
    Methods in molecular biology (Clifton, N.J.) 08/2012; 916:357-72. DOI:10.1007/978-1-61779-980-8_26 · 1.29 Impact Factor
  • Graziella Pellegrini · Paolo Rama · Michele De Luca ·
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    ABSTRACT: Cultures of limbal cells are a safe and effective treatment for the destruction of the human cornea owing to chemical burns. The essential feature of the graft is the presence of an adequate number of stem cells, which can be determined by the expression of the p63 transcription factor. Here, we will discuss the general principles defining the rigorous criteria for graftable limbal cultures in light of their clinical performances. Such criteria might prove relevant to the future therapeutic use of any cultured cell type.
    Trends in Molecular Medicine 11/2010; 17(1). DOI:10.1016/j.molmed.2010.10.003 · 9.45 Impact Factor
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    ABSTRACT: Corneal renewal and repair are mediated by stem cells of the limbus, the narrow zone between the cornea and the bulbar conjunctiva. Ocular burns may destroy the limbus, causing limbal stem-cell deficiency. We investigated the long-term clinical results of cell therapy in patients with burn-related corneal destruction associated with limbal stem-cell deficiency, a highly disabling ocular disease. We used autologous limbal stem cells cultivated on fibrin to treat 112 patients with corneal damage, most of whom had burn-dependent limbal stem-cell deficiency. Clinical results were assessed by means of Kaplan-Meier, Kruskal-Wallis, and univariate and multivariate logistic-regression analyses. We also assessed the clinical outcome according to the percentage of holoclone-forming stem cells, detected as cells that stain intensely (p63-bright cells) in the cultures. Permanent restoration of a transparent, renewing corneal epithelium was attained in 76.6% of eyes. The failures occurred within the first year. Restored eyes remained stable over time, with up to 10 years of follow-up (mean, 2.91+/-1.99; median, 1.93). In post hoc analyses, success--that is, the generation of normal epithelium on donor stroma--was associated with the percentage of p63-bright holoclone-forming stem cells in culture. Cultures in which p63-bright cells constituted more than 3% of the total number of clonogenic cells were associated with successful transplantation in 78% of patients. In contrast, cultures in which such cells made up 3% or less of the total number of cells were associated with successful transplantation in only 11% of patients. Graft failure was also associated with the type of initial ocular damage and postoperative complications. Cultures of limbal stem cells represent a source of cells for transplantation in the treatment of destruction of the human cornea due to burns.
    New England Journal of Medicine 07/2010; 363(2):147-55. DOI:10.1056/NEJMoa0905955 · 55.87 Impact Factor
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    Graziella Pellegrini · Michele De Luca ·
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    ABSTRACT: Recently in The Lancet, Guenou et al. (2009) demonstrate that human embryonic stem cells (hESCs) can differentiate into mature keratinocytes able to generate a pluristratified epithelium on immunodeficient mice. Their findings and the potential clinical use of hESC-derived keratinocytes will be discussed.
    Cell stem cell 01/2010; 6(1):8-9. DOI:10.1016/j.stem.2009.12.002 · 22.27 Impact Factor
  • M De Luca · G Pellegrini · F Mavilio ·
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    ABSTRACT: Gene therapy has the potential to treat devastating inherited diseases for which there is little hope of finding a conventional cure. These include lethal diseases, like immunodeficiencies or several metabolic disorders, or conditions associated with a relatively long life expectancy but poor quality of life and expensive and life-long symptomatic treatments, such as muscular dystrophy, cystic fibrosis and thalassaemia. Skin adhesion defects belong to both groups. For the nonlethal forms, gene therapy, or transplantation of cultured skin derived from genetically corrected epidermal stem cells, represents a very attractive therapeutic option, and potentially a definitive treatment. Recent advances in gene transfer and stem cell culture technology are making this option closer than ever. This paper critically reviews the progress and prospects of gene therapy for epidermolysis bullosa, and the technical and nontechnical factors currently limiting its development.
    British Journal of Dermatology 06/2009; 161(1):19-24. DOI:10.1111/j.1365-2133.2009.09243.x · 4.28 Impact Factor
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    ABSTRACT: Mucins released into the tear film are crucial to maintaining a healthy ocular surface. Alterations in goblet cell numbers and mucin secretion are observed in chronic ocular surface inflammatory diseases. Nerve growth factor (NGF) plays a crucial role in healing and inflammation of the ocular surface. The aim of this study was to evaluate in vitro the effect of NGF on conjunctival goblet cell differentiation and mucin production and secretion. Human conjunctival epithelial cells were exposed to increasing NGF concentrations (1 to 250 ng/mL) and analyzed to quantify cell growth (MTT/Ki67/BrdU), goblet cell differentiation (PAS/MUC5AC confocal staining), and mucin mRNA expression (real-time PCR). Secreted and cellular MUC5AC were also analyzed by sandwich-ELISA and FACS, respectively. To confirm the biological effects of NGF, the same evaluations were performed on primary cultures, and changes in markers of stemness (p63) and commitment (14-3-3 sigma) were also investigated. In cell cultures, NGF induced a dose-dependent increase of goblet cell numbers, MUC5AC production, storage, and release. Additionally, in primary cultures, NGF induced an increase of abortive colonies and 14-3-3 sigma protein, and a decrease of p63 mRNA and protein, suggesting a differentiating effect of NGF on human conjunctival epithelium. These findings show that NGF might play a role in the complex mechanism leading to conjunctival epithelium differentiation and mucin secretion. In addition to the known roles of NGF in promoting ocular surface healing and sensitivity, its effects on conjunctival goblet cells support a rationale to investigate the therapeutic effectiveness of NGF in dry eye disease.
    Investigative ophthalmology & visual science 05/2009; 50(10):4622-30. DOI:10.1167/iovs.08-2716 · 3.40 Impact Factor
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    ABSTRACT: Considerable practical hurdles must be overcome prior to the broad application of stem cell therapies. We outline challenges that may vary across different models of cell therapy, including the following broad concepts: issues related to the sourcing of material, and issues related to product manufacturing, shipping, storage and tracking, and standardization.
    Cell stem cell 01/2009; 4(1):20-6. DOI:10.1016/j.stem.2008.11.012 · 22.27 Impact Factor
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    ABSTRACT: The International Society for Stem Cell Research (ISSCR) task force that developed new Guidelines for the Clinical Translation of Stem Cells discusses core principles that should guide the responsible transition of basic stem cell research into appropriate clinical applications.
    Cell stem cell 01/2009; 3(6):607-9. DOI:10.1016/j.stem.2008.11.009 · 22.27 Impact Factor
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    G Pellegrini · P Rama · F Mavilio · M De Luca ·
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    ABSTRACT: Regenerative medicine refers to innovative therapies aimed at the permanent restoration of diseased tissues and organs. Regeneration of self-renewing tissues requires specific adult stem cells, which need to be genetically modified to correct inherited genetic diseases. Cultures of epithelial stem cells permanently restore severe skin and mucosal defects, and genetically corrected epidermal stem cells regenerate a normal epidermis in patients carrying junctional epidermolysis bullosa. The keratinocyte stem cell is therefore the only cultured stem cell used both in cell therapy and gene therapy clinical protocols. Epithelial stem cell identification, fate and molecular phenotype have been extensively reviewed, but not in relation to tissue regeneration. In this paper we focus on the localization and molecular characterization of human limbal stem cells in relation to corneal regeneration, and the gene therapy of genetic skin diseases by means of genetically modified epidermal stem cells.
    The Journal of Pathology 01/2009; 217(2):217-28. DOI:10.1002/path.2441 · 7.43 Impact Factor

Publication Stats

8k Citations
891.11 Total Impact Points


  • 2005-2015
    • Università degli Studi di Modena e Reggio Emilia
      • • Center for Regenerative Medicine "Stefano Ferrari"
      • • Department of Life Sciences
      Modène, Emilia-Romagna, Italy
  • 2010
    • San Raffaele Scientific Institute
      Milano, Lombardy, Italy
  • 2006
    • The American University of Rome
      Roma, Latium, Italy
  • 2004
    • Banca d'Italia
      Roma, Latium, Italy
  • 2002
    • Harvard Medical School
      Boston, Massachusetts, United States
  • 1997-2002
    • Istituto Dermopatico dell'Immacolata
      Roma, Latium, Italy
    • Università degli Studi di Siena
      Siena, Tuscany, Italy
  • 1999-2001
    • University of Rome Tor Vergata
      Roma, Latium, Italy
  • 1998
    • Sapienza University of Rome
      Roma, Latium, Italy
  • 1994
    • Biotecnologie Avanzate
      Napoli, Campania, Italy
  • 1993
    • Massachusetts General Hospital
      Boston, Massachusetts, United States
  • 1990-1992
    • Università degli Studi di Torino
      Torino, Piedmont, Italy
  • 1988-1992
    • Università degli Studi di Genova
      • Dipartimento di Medicina sperimentale (DIMES)
      Genova, Liguria, Italy
  • 1991
    • University of Bologna
      Bolonia, Emilia-Romagna, Italy