Cell-Based Therapy for Dystrophic Epidermolysis Bullosa

营养不良性大疱性表皮松解症的细胞疗法

• 批准号: 7532583
• 负责人: OLGA IGOUCHEVA
• 金额: $ 20.39万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7532583
• 关键词: Adhesions; Affect; Bandage; Basement membrane; Bone Marrow; Bone and Cartilage Funding; Cell Therapy; Cell Transplants; Cells; Cessation of life; Clinical; Collagen; Collagen Type IV; Collagen Type VII; Condition; Connective Tissue; Connective Tissue Diseases; Deposition; Dermal; Dermis; Development; Disease; Disease Management; Engraftment; Environment; Epidermolysis Bullosa; Epidermolysis Bullosa Dystrophica; Epithelial; Epithelium; Fatty acid glycerol esters; Fiber; Fibroblasts; Gene Mutation; Genes; Genetic; Hereditary Disease; Human; In Vitro; Infection; Inherited; Kinetics; Knowledge; Laminin; Lead; Lesion; Marrow; Mechanics; Mesenchymal; Mesenchymal Stem Cells; Mesenchyme; Metabolic; Mucous Membrane; Mus; Muscle; Mutation; Numbers; Outcome Study; Papillary; Pathologic; Pathology; Patients; Phenotype; Prevention; Property; Proteins; Public Health; Research; Route; Skin; Skin Cancer; Squamous cell carcinoma; Stem Cell Research; Stimulus; Structure; Tendon structure; Testing; Therapeutic; Tissues; Topical Antibiotic; Translating; Transplantation; Trauma; Work; adult stem cell; day; disease phenotype; keratinocyte; lamina densa; macromolecule; mouse model; non-viral gene therapy; novel therapeutics; restoration; skin disorder; therapeutic protein

DESCRIPTION (provided by applicant): Dystrophic epidermolysis bullosa (DEB) is an inherited mechanobullous disorder characterized by fragility of the skin and mucous membranes. The morphological hallmark of DEB includes splitting of the basement membrane zone of the dermal-epidermal junction, with a cleavage plane lying within the papillary dermis. The abnormal assembly of anchoring fibrils is caused by genetic mutations in the gene (COL7A1) encoding collagen type VII synthesized by the basal keratinocytes of the epithelium and the fibroblast of the papillary dermis. At present, there are no specific therapies for any form of inherited DEB. Day-to-day management of the disease is concentrated on the prevention of mechanical trauma, infections via judicious use of bandages or loose-fitting garments and topical antibiotics. Precise understanding of the genetic lesions underlying different subtypes of DEB, and elucidation of the consequences of such mutations at the protein level, enabled us to continue our attempts to develop therapy approaches towards counteracting the clinical manifestations in this devastating skin disease. Recent advances in stem cell research have raised the possibility that use of adult stem cells from bone marrow may provide dramatic new therapies for treatment of inherited and acquired diseases. Bone marrow contains mesenchymal progenitor cells, called mesenchymal stem cells (MSC), that have several unique properties suggesting the feasibility of their use as a therapeutic vehicle: (1) a relatively simple isolation; (2) the ability to be cultured in vitro in minimal conditions and to expand to quantities required for therapy; (3) the ability for ex vivo transduction; (4) plasticity, the potential to differentiate under exogenous stimuli; (5) the ability to engraft after reintroduction; (6) high metabolic activity and efficient machinery to express therapeutic proteins in secretory form; and, (7) the ability to be delivered systematically or locally. The proposed research is a logistical extension of our work in the field of non-viral gene therapy. The main objective of these studies is to test the hypothesis that engraftment of genetically normal MSC will lead to expansion of the transplanted cells in sufficient numbers in the skin of DEB mice and the level of collagen VII will be sufficient to reduce the pathologic phenotype. The following parameters are will be investigated: (1) the rout of MSC delivery, distribution and quantity of transplanted cells in DEB skin; (2) the kinetics of MSC distribution in DEB skin; (3) the phenotypic and functional features of engrafted MSC; (4) the kinetics of distribution and release of mature collagen VII in the intact mouse skin; (5) the ability of MSC to produce collagen VII to form anchoring fibril-like structures; (6) the deposition of collagen VII fibers into the BMZ; (7) the structure of the newly formed collagen VII fibers; and, (8) the ability of newly formed fibers to interact with BMZ's macromolecules. A successful outcome of these studies may form a platform for development of a novel therapeutic approach for curing connective tissue disorders, as well as other human genetic disorders. In the current project, we suggested to test the hypothesis that engraftment of genetically normal mesenchymal stem cells isolated from bone marrow will lead to expansion of transplanted cells in sufficient numbers in the affected skin of DEB mice and the level of collagen VII will be sufficient to reduce the pathologic phenotype. If successful, the suggested strategy may become powerful therapeutic approach for treatment of connective tissue disorders, in particular dystrophic epidermolysis bullosa, and benefit public health.

描述（由申请方提供）：营养不良性大疱性表皮病（DEB）是一种遗传性机械性大疱性疾病，其特征为皮肤和粘膜脆弱。DEB的形态学特征包括真皮-表皮交界处的基底膜区分裂，其中分裂平面位于乳头状真皮内。锚定原纤维的异常组装是由编码VII型胶原的基因（COL 7A 1）中的基因突变引起的，所述VII型胶原由上皮的基底角质形成细胞和乳头状真皮的成纤维细胞合成。目前，对于任何形式的遗传性DEB都没有特定的治疗方法。该疾病的日常管理集中在通过明智地使用绷带或宽松的衣服和局部抗生素来预防机械创伤、感染。对DEB不同亚型的遗传病变的精确理解，以及对这种突变在蛋白质水平上的后果的阐明，使我们能够继续尝试开发治疗方法，以抵消这种毁灭性皮肤病的临床表现。干细胞研究的最新进展提高了使用来自骨髓的成体干细胞可能为治疗遗传性和获得性疾病提供引人注目的新疗法的可能性。骨髓含有间充质祖细胞，称为间充质干细胞（MSC），其具有几种独特的性质，表明其用作治疗载体的可行性：（1）相对简单的分离;（2）在最低条件下体外培养并扩增至治疗所需量的能力;（3）离体转导的能力;（4）可塑性，即在外源性刺激下分化的潜力;（5）在重新引入后移植的能力;（6）高代谢活性和以分泌形式表达治疗性蛋白质的有效机制;以及（7）系统或局部递送的能力。拟议的研究是我们在非病毒基因治疗领域工作的后勤延伸。这些研究的主要目的是检验以下假设：植入遗传正常的MSC将导致DEB小鼠皮肤中足够数量的移植细胞扩增，并且胶原蛋白VII的水平将足以降低病理表型。本研究将研究以下参数：（1）MSC的递送途径、在DEB皮肤中移植细胞的分布和数量;（2）MSC在DEB皮肤中的分布动力学;（3）移植MSC的表型和功能特征;（4）成熟胶原VII在完整小鼠皮肤中的分布和释放动力学;（5）MSC在DEB皮肤中的表型和功能特征。（5）MSC产生胶原VII以形成锚定纤维样结构的能力;（6）胶原VII纤维沉积到BMZ中;（7）新形成的胶原VII纤维的结构;和（8）新形成的纤维与BMZ的大分子相互作用的能力。这些研究的成功结果可能为开发治疗结缔组织疾病以及其他人类遗传疾病的新治疗方法提供平台。在目前的项目中，我们建议测试以下假设：从骨髓中分离的遗传正常的间充质干细胞的植入将导致在DEB小鼠的受影响皮肤中足够数量的移植细胞的扩增，并且胶原蛋白VII的水平将足以减少病理表型。如果成功，建议的策略可能成为治疗结缔组织疾病，特别是营养不良性大疱性表皮病的有力治疗方法，并有益于公众健康。