Gene Transfer for Recessive Dystrophic Epidermolysis Bullosa

描述（由申请人提供）：大疱性表皮病（EB）是一个遗传性遗传性起泡性皮肤病家族。缺乏正常VII型胶原蛋白的儿童发展为严重的、瘢痕性EB亚型，隐性营养不良性大疱性表皮病（RDEB），其在皮肤和粘膜上产生疼痛的水泡和伤口。RDEB受试者的预期寿命缩短，并因感染、器官衰竭或鳞状细胞癌（SCC）而过早死亡。目前RDEB的治疗仅限于姑息性伤口护理，因为没有改变疾病病程或严重程度的治疗方法。我们专注于开发RDEB的分子疗法。我们已经证明，基因校正的RDEB角质形成细胞工程表达VII型胶原蛋白可以纠正人RDEB皮肤组织移植到免疫缺陷小鼠。我们现在计划将这种方法扩展到RDEB受试者，通过在人类基因转移临床试验中将遗传校正的自体RDEB角质形成细胞移植回RDEB受试者的皮肤中。本申请的具体目的是将VII型胶原蛋白递送至RDEB受试者的皮肤，并评价有益效果的有效性和持续时间。该R01申请记录了我们之前在动物模型方面的长期成功，我们的基因转移技术开发，以及我们目前为1期临床试验确定和招募受试者的计划。成功的基因转移将产生VII型胶原蛋白，其正确定位于皮肤BMZ，并掺入超微结构正常的锚定原纤维中。我们将评估VII型胶原蛋白在基因组保留水平以及mRNA和蛋白质表达水平上的持久性。我们将评估原始皮肤和基因矫正皮肤的基因表达谱。我们将评估移植皮肤对递送的VII型胶原蛋白的不必要的免疫应答。对遗传校正的自体皮肤移植物的RDEB受体中的任何体液或细胞免疫应答的分析将提供关于这种方法成功的可能性以及对任何耐久性失败的洞察的重要信息。该应用通过转化应用先前的基础科学成功，通过对这种严重皮肤病的治疗进行集中研究，实现了国家关节炎、肌肉骨骼和皮肤病研究所的使命。我们希望这项试验可能会为这些受试者带来一种潜在有效的基于细胞的治疗方法，并可能进一步支持遗传性皮肤病的矫正基因治疗。公共卫生相关性：本申请提出开发用于称为隐性营养不良性大疱性表皮病（RDEB）的严重皮肤病的基因疗法。我们计划纠正受影响的人自己的皮肤细胞，并将他们自己的细胞移植回皮肤上。这种皮肤病的有效基因治疗将有助于其他遗传性疾病基因治疗的发展。

DESCRIPTION (provided by applicant): Epidermolysis bullosa (EB) is a family of inherited genetic blistering skin disorders. Children lacking normal type VII collagen develop a severe, scarring EB subtype, recessive dystrophic epidermolysis bullosa (RDEB), which produces painful blisters and wounds on skin and mucous membranes. RDEB subjects have a shortened life expectancy with early death from infection, organ failure or squamous cell carcinoma (SCC). Current therapy for RDEB is limited to palliative wound care as there are no therapies available that alter the course or severity of the disease. We have focused on developing molecular therapy for RDEB. We have demonstrated that genetically corrected RDEB keratinocytes engineered to express type VII collagen can correct human RDEB skin tissue grafted onto immune deficient mice. We now plan to extend this approach to RDEB subjects by grafting genetically corrected autologous RDEB keratinocytes back into the skin of RDEB subjects in a human gene transfer clinical trial. The Specific Aims of this application are to deliver type VII collagen to the skin of RDEB subjects and to evaluate the effectiveness and duration of the beneficial effect. This R01 application documents our previous long term success in animal models, our gene transfer technology development, and our current plan to identify and enroll subjects for a Phase 1 clinic trial. Successful gene transfer will produce type VII collagen protein correctly localized to the cutaneous BMZ that is incorporated into ultrastructurally normal anchoring fibrils. We will assess the durability of type VII collagen at the levels of genomic retention, as well as mRNA and protein expression. We will evaluate gene expression profiles of the original and the gene corrected skin. We will evaluate grafted skin for unwanted immune response to the delivered type VII collagen protein. Analysis for any humoral or cellular immune responses in RDEB recipients of genetically corrected autologous skin grafts will provide important information both about the likelihood of success of this approach as well as insight into any failure of durability. This application fulfills the mission of the National Institute of Arthritis and Musculoskeletal and Skin Diseases through focused research upon treatment of this severe skin disease by translational application of previous basic science successes. We expect that this trial may lead to a potentially effective cell based therapy for these subjects and may further support corrective gene therapy for genetic skin diseases. PUBLIC HEALTH RELEVANCE: This application proposes to develop gene therapy for a severe skin disease called recessive dystrophic epidermolysis bullosa (RDEB). We plan to correct the affected person's own skin cells and transplant their own cells back onto their skin. Effective gene therapy in this skin disease will assist in development of gene therapy for other genetic diseases.