title: Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB.
abstract: Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen, a protein essential for anchoring fibril formation at the dermal-epidermal junction. Whereas allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection and support sustained delivery of type VII collagen at the dermal-epidermal junction. We demonstrate the feasibility of such an approach using a therapeutic grade, self-inactivating-lentiviral vector, encoding codon-optimized COL7A1, to transduce RDEB fibroblasts under conditions suitable for clinical application. Expression and secretion of type VII collagen was confirmed with transduced cells exhibiting supranormal levels of protein expression, and ex vivo migration of fibroblasts was restored in functional assays. Gene-modified RDEB fibroblasts also deposited type VII collagen at the dermal-epidermal junction of human RDEB skin xenografts placed on NOD-scid IL2Rgamma(null) recipients, with reconstruction of human epidermal structure and regeneration of anchoring fibrils at the dermal-epidermal junction. Fibroblast-mediated restoration of protein and structural defects in this RDEB model strongly supports proposed therapeutic applications in man.
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom.
authors.forename: Christos
authors.initials: C
authors.lastname: Georgiadis
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom.
authors.forename: Farhatullah
authors.initials: F
authors.lastname: Syed
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Petrova
authors.affiliation: St John's Institute of Dermatology, King's College London (Guy's campus), London, United Kingdom.
authors.forename: Alya
authors.initials: A
authors.lastname: Abdul-Wahab
authors.affiliation: St John's Institute of Dermatology, King's College London (Guy's campus), London, United Kingdom.
authors.forename: Su M
authors.initials: SM
authors.lastname: Lwin
authors.affiliation: Department of Haematological Medicine, King's College London, The Rayne Institute, London, United Kingdom.
authors.forename: Farzin
authors.initials: F
authors.lastname: Farzaneh
authors.affiliation: Department of Haematological Medicine, King's College London, The Rayne Institute, London, United Kingdom.
authors.forename: Lucas
authors.initials: L
authors.lastname: Chan
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom.
authors.forename: Sumera
authors.initials: S
authors.lastname: Ghani
authors.affiliation: Centre for Ultrastructural Imaging, King's College London, London, United Kingdom.
authors.forename: Roland A
authors.initials: RA
authors.lastname: Fleck
authors.affiliation: Centre for Ultrastructural Imaging, King's College London, London, United Kingdom.
authors.forename: Leanne
authors.initials: L
authors.lastname: Glover
authors.affiliation: The Robin Eady National Diagnostic Epidermolysis Bullosa Laboratory, Viapath LLP, St Thomas' Hospital, London, United Kingdom.
authors.forename: James R
authors.initials: JR
authors.lastname: McMillan
authors.affiliation: Department of Dermatology, University of Southern California, Los Angeles, California, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Chen
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom.
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.affiliation: St John's Institute of Dermatology, King's College London (Guy's campus), London, United Kingdom.
authors.forename: John A
authors.initials: JA
authors.lastname: McGrath
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom.
authors.forename: Wei-Li
authors.initials: WL
authors.lastname: Di
authors.affiliation: UCL Institute of Child Health, Molecular and Cellular Immunology Section & Great Ormond Street Hospital NHS Foundation Trust, London, United Kingdom. Electronic address: W.Qasim@ucl.ac.uk.
authors.forename: Waseem
authors.initials: W
authors.lastname: Qasim
mesh_terms: D000818:Animals; D003062:Codon; D024143:Collagen Type VII / Q000235:genetics*; D004195:Disease Models, Animal; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics* / Q000628:therapy*; D005347:Fibroblasts / Q000637:transplantation*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D064593:Heterografts; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008297:Male; D051379:Mice; D016513:Mice, SCID; D011897:Random Allocation; D016038:Skin Transplantation / Q000379:methods; D023822:Tissue Engineering; D014945:Wound Healing / Q000502:physiology
chemical_list: D003062:Codon; C583886:Col7a1 protein, mouse; D024143:Collagen Type VII