Remington Jennifer, Wang Xinyi, Hou Yingpin, Zhou Hui, Burnett Julie, Muirhead Trevor, Uitto Jouni, Keene Douglas R, Woodley David T, Chen Mei
Department of Dermatology, University of Southern California, Los Angeles, California, USA.
Mol Ther. 2009 Jan;17(1):26-33. doi: 10.1038/mt.2008.234. Epub 2008 Nov 18.
Patients with recessive dystrophic epidermolysis bullosa (RDEB) have incurable skin fragility, blistering, and scarring due to mutations in the gene that encodes for type VII collagen (C7) that mediates dermal-epidermal adherence in human skin. We showed previously that intradermal injection of recombinant C7 into transplanted human DEB skin equivalents stably restored C7 expression at the basement membrane zone (BMZ) and reversed the RDEB disease features. In this study, we evaluated the feasibility of protein therapy in a C7 null mouse (Col7a1(-/-)) which recapitulates the features of human RDEB. We intradermally injected purified human C7 into DEB mice and found that the injected human C7 stably incorporated into the mouse BMZ, formed anchoring fibrils, and corrected the DEB murine phenotype, as demonstrated by decreased skin fragility, reduced new blister formation, and markedly prolonged survival. After 4 weeks, treated DEB mice developed circulating anti-human C7 antibodies. Most surprisingly, these anti-C7 antibodies neither bound directly to the mouse's BMZ nor prevented the incorporation of newly injected human C7 into the BMZ. Anti-C7 antibody production was prevented by treating the mice with an anti-CD40L monoclonal antibody, MR1. We conclude that protein therapy may be feasible for the treatment of human patients with RDEB.
隐性营养不良性大疱性表皮松解症(RDEB)患者因编码VII型胶原蛋白(C7)的基因突变而患有无法治愈的皮肤脆弱、水疱形成和瘢痕形成,C7在人皮肤中介导真皮-表皮黏附。我们之前表明,将重组C7皮内注射到移植的人DEB皮肤等效物中可在基底膜带(BMZ)稳定恢复C7表达并逆转RDEB疾病特征。在本研究中,我们评估了在重现人类RDEB特征的C7基因敲除小鼠(Col7a1(-/-))中进行蛋白质治疗的可行性。我们将纯化的人C7皮内注射到DEB小鼠中,发现注射的人C7稳定整合到小鼠BMZ中,形成锚定原纤维,并纠正了DEB小鼠的表型,表现为皮肤脆弱性降低、新水疱形成减少和生存期显著延长。4周后,接受治疗的DEB小鼠产生了循环抗人C7抗体。最令人惊讶的是,这些抗C7抗体既不直接结合小鼠的BMZ,也不阻止新注射的人C7整合到BMZ中。用抗CD40L单克隆抗体MR1治疗小鼠可防止抗C7抗体的产生。我们得出结论,蛋白质治疗对于治疗人类RDEB患者可能是可行的。