Skin-targeted Cell Therapy for Recessive Dystrophic Epidermolysis Bullosa

隐性营养不良性大疱性表皮松解症的皮肤靶向细胞疗法

基本信息

批准号：
10490837
负责人：
Jakub Tolar
金额：
$ 33.76万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10490837
关键词：
Address Affect Anemia Apoptosis Area Autoimmunity Autologous Binding Biodistribution Biological Bulla Burn injury COL7A1 Cell Lineage Cell Therapy Cells Chemicals Clustered Regularly Interspaced Short Palindromic Repeats Collagen Gene Collagen Type VII Complex Contracture Corneal Abrasion DNA DNA Damage Data Deaminase Deoxyribonucleases Disease Engraftment Epidermolysis Bullosa Dystrophica Esophageal Stenosis Family Fibroblasts Fibrosis Gene Mutation Gene Transfer Genes Genetic Diseases Genome Genotoxic Stress Goals Human Immune response Individual Inflammation Inflammatory Response Joints Laboratories Laboratory Study Lead Longevity Malignant - descriptor Malnutrition Measures Mediating Membrane Membrane Proteins Mesenchymal Modeling Molecular Mucous Membrane Mus Mutation Nature Nucleotides Patients Persons Phenotype Pre-Clinical Model Production Proteins Reagent Recording of previous events Research Rest Risk Side Site Skin Skin injury Skin repair Source Squamous cell carcinoma Stress Stromal Cells Technology Testing Therapeutic Therapeutic Intervention Toxic effect Tropism Ulcer Work Xenograft procedure base base editing biological systems cell type clinical application clinically relevant combinatorial effective therapy gene correction gene therapy genotoxicity healing improved in vivo evaluation induced pluripotent stem cell insight keratinocyte loss of function mutation member mesenchymal stromal cell molecular pathology mouse model next generation novel nuclease operation phosphodiester polymerization pre-clinical programs prototype response restoration skin regeneration stem cells tool vector wound wound healing

项目摘要

Abstract Recessive dystrophic epidermolysis bullosa (RDEB) is a prototypical genodermatosis caused by biallelic loss- of-function mutations of COL7A1. These mutations lead to a lack of type VII collagen (C7) in the skin and mucosal membranes, resulting in a complex phenotype of blistering, fibrosis, pseudosyndactyly, joint contractures, esophageal strictures, corneal abrasions, malnutrition, autoimmunity, anemia, and squamous cell carcinoma. Despite tremendous efforts over the last decade to establish curative measures for this severe and potentially fatal disorder, there are as yet no therapies that reliably supply C7 protein to the multiple sites affected by generalized severe RDEB. To address this, we propose to gain a more mechanistic understanding of how to restore the integrity of COL7A1 without causing collateral damage to the rest of the genome, and of how the specialized tropism of cells works to deliver intact, functional C7 throughout the body. In order to accomplish these goals and to overcome the limitations of current gene and cell therapies, we will investigate the following questions: [i] Is base editing superior to CRISPR/Cas9-editing for correction of COL7A1 mutations? Because base editing does not cause double-strand breaks in the way that classic gene editing with DNA nucleases does, it avoids genotoxic stress. [ii] Are skin-specialized cells, such as ABCB5+ mesenchymal stromal/stem cells (MSCs), superior to alternative sources of MSCs in expression of C7 levels adequate for cross-correction of C7 deficiency in RDEB? We will evaluate skin-specific stromal cells, such as mesenchymal stromal cells expressing ATP-binding cassette sub-family B member 5 (ABCB5+) surface protein, derived directly from skin or indirectly from patient-specific induced pluripotent stem cells, which have had COL7A1 restored to function with base editing. [iii] Do COL7A1-edited human ABCB5+ MSCs mediate wound healing in a preclinical murine model of RDEB? Using our murine model of RDEB that accepts human xenografts, we will quantify the value of base editing-corrected ABCB5+ MSCs and induced pluripotent stem cell-derived MSCs. We propose to define the conditions conducive to wound healing in this severe blistering genodermatosis by using powerful tools for studying and manipulating the information bases of biological systems (i.e., programmable deaminases for base editing-mediated gene therapy; induced cell lineage conversion; and skin tropism). We will aim for personalized cell therapy for individuals with generalized severe RDEB, with the idea that our findings may provide insights into ways to manage other genodermatoses, as well as treatment of mucocutaneous ulcers, and chemical and thermal burns. Our proposal is equally motived by wanting a better understanding of the biological mechanisms in injured skin and by needing to improve the lives of people with RDEB through reducing the risks and maximizing the benefits of potential novel gene and cell therapies.

抽象的隐性营养不良的表皮溶解Bullosa（RDEB）是由双重损失引起的典型的胚芽病。 Col7a1的功能突变。这些突变导致皮肤中缺乏VII型胶原蛋白（C7），并且粘膜膜，导致起泡，纤维化，伪合成，关节的复杂表型染色，食管狭窄，角膜擦伤，营养不良，自身免疫性，贫血和鳞状细胞癌。尽管在过去十年中做出了巨大努力，以为这种严重和潜在的致命疾病，目前尚无可靠地向多个部位提供C7蛋白的疗法受广义严重RDEB的影响。为了解决这个问题，我们建议对如何恢复COL7A1的完整性，而不会对基因组的其余部分造成附带损害，以及如何细胞的专门向热门作用，可以在整个体内提供完整的功能性C7。为了实现这些目标并克服当前基因和细胞疗法的局限性，我们将调查以下问题：[i]基础编辑优于CRISPR/CAS9编辑以纠正COL7A1 突变？因为基础编辑不会引起双链断裂的方式，而经典基因编辑的方式 DNA核酸酶确实可以避免遗传毒性应激。 [ii]是皮肤专门的细胞，例如ABCB5+间充质基质/干细胞（MSC），在C7水平表达中优于MSC的替代来源，足以适合 RDEB中C7缺乏的跨校正？我们将评估皮肤特异性基质细胞，例如间充质表达ATP结合盒子子家庭B成员5（ABCB5+）表面蛋白的基质细胞，衍生直接来自皮肤或间接来自患者特异性诱导的多能干细胞，这些干细胞具有COL7A1 恢复以通过基础编辑功能。 [iii] DO COL7A1编辑的人ABCB5+ MSC介导伤口愈合 RDEB的临床前鼠模型？使用我们接受人类异种移植的鼠模型的鼠模型，我们将量化基本编辑校正的ABCB5+ MSC和诱导多能干细胞衍生的MSC的值。我们建议在这种严重的泡沫性GETERMATOSIS中定义有利于伤口愈合的疾病使用强大的工具来研究和操纵生物系统的信息库（即用于基础编辑介导的基因疗法的可编程脱氨酶；诱导细胞谱系转换；和皮肤向性）。我们将针对具有广义重度RDEB的个体的个性化细胞疗法，并有这个想法我们的发现可能会提供有关管理其他Genedermatoses的方法的见解，以及粘膜溃疡以及化学和热燃烧。我们的建议同样希望更好了解受伤皮肤中的生物学机制，并需要改善患者的生活 RDEB通过降低风险并最大程度地提高潜在的新型基因和细胞疗法的益处。