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Single Step Strategy for the Therapeutic Reprogramming in Epidermolysis Bullosa Patients

大疱性表皮松解症患者治疗重编程的一步策略

基本信息

批准号：
9317254
负责人：
Vittorio Sebastiano
金额：
$ 20.72万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-05-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9317254
关键词：
Abnormal Karyotype Address Affect Age Agreement Area Autologous Blindness Bone Marrow Transplantation Bulla CRISPR/Cas technology Cell Line Cell Therapy Cells Chemicals Clinic Clinical Clinical Trials Clone Cells Cloning Collagen Type VII Data Degenerative Disorder Derivation procedure Dermal Development Disease Epidermolysis Bullosa Epidermolysis Bullosa Dystrophica Fibroblasts Frequencies Gene Targeting Generations Genetic Genome Genomic DNA Goals Guide RNA Hereditary Disease Human Immunocompromised Host Impaired wound healing In Situ In Vitro Incidence Individual Lead Life Malignant Neoplasms Medical Messenger RNA Modification Mus Mutation National Institute of Arthritis and Musculoskeletal and Skin Diseases Nature Other Genetics Patients Physiological Population Proteins Protocols documentation Public Health Publishing RNA Sequences Reagent Regenerative Medicine Risk Safety Skin Somatic Cell Somatic Gene Therapy Somatic Mutation Source Squamous cell carcinoma Stem cells Surface Susceptibility Gene Technology Testing Therapeutic Time Tissues Translations Transplantation Tumorigenicity Undifferentiated United States National Institutes of Health Work alternative treatment base cell bank cell type clinical application clinically translatable cytotoxicity design exhaustion exome sequencing experience functional restoration gene correction genome editing genome sequencing homologous recombination improved in vivo induced pluripotent stem cell innovation keratinocyte novel novel strategies success tissue culture tissue regeneration tool virtual whole genome

项目摘要

The use of human induced Pluripotent Stem Cells (iPSCs) for regenerative medicine is an attractive alternative for the treatment of genetic and degenerative diseases in which an extensive or continuous need of tissue regeneration is required. iPSCs are more easily amenable to in situ gene correction and can be expanded indefinitely retaining a pluripotent and undifferentiated state, and can therefore be used as a constant source of material for cell therapy. Development of iPSC-based therapies for Recessive Dystrophic Epidermolysis Bullosa (RDEB) patients represents an ideal paradigm in this scenario owing to the severe nature of the disease, the demonstration that corrected keratinocytes can have long-term tissue repopulation, and the need for large numbers of stem cells to cover the affected surface area. We have recently demonstrated that Therapeutic Reprogramming (TR) (achieved by genetic correction of RDEB patients-derived iPSC) provides a virtually unlimited source of genetically corrected autologous keratinocytes that can from multi-layered skin with restored functionality and physiological expression of Collagen VII type 1 (Sebastiano et al., 2014). While effective, one of the major bottlenecks of this approach is the many sub-cloning steps required to derive the corrected iPSC clones, with an inherent accumulation of a large number of somatic mutations and karyotype abnormalities (in agreement with a conspicuous body of evidence in the field) that could have unpredictable effects in vivo after transplantation and with serious implications about the safety of iPSC-based therapy in general. This represents a fundamental problem and a preeminent priority that needs to be addressed before translation of iPSC-based technologies to the clinic is put in place. In this proposal we are aiming at tackling this problem by proposing a novel approach to derive genetically corrected iPSCs carrying a slim number of somatic mutations. We believe our proposal will have a fundamental impact on the safety of iPSC-based therapies and will serve as a paradigm for the development of release criteria that will be used to qualify iPSC-banks for any genetic and degenerative disease.

使用人类诱导多能干细胞 (iPSC) 进行再生药物是治疗遗传性和退行性疾病的一种有吸引力的替代方案广泛或持续需要组织再生的疾病必需的。 iPSC 更容易进行原位基因校正，并且可以无限期地扩增，保留多能和未分化状态，并且可以因此可用作细胞治疗的恒定材料来源。发展基于 iPSC 的隐性营养不良性大疱性表皮松解症 (RDEB) 疗法由于其严重性，患者代表了这种情况下的理想范例的疾病，证明纠正角质形成细胞可以长期组织再生，以及需要大量干细胞来覆盖受影响的表面积。我们最近证明了治疗重编程（TR）（通过 RDEB 患者衍生的基因校正实现 iPSC）提供了几乎无限的基因校正自体细胞来源角质形成细胞可以来自具有恢复功能的多层皮肤， VII 型胶原蛋白 1 的生理表达（Sebastiano 等人，2014）。尽管有效，这种方法的主要瓶颈之一是许多子克隆衍生具有固有积累的校正 iPSC 克隆所需的步骤大量体细胞突变和核型异常（在与该领域大量明显的证据达成一致）移植后体内不可预测的影响并具有严重影响关于基于 iPSC 的治疗的总体安全性。这代表了一个基本的翻译前需要解决的问题和首要优先事项基于 iPSC 的技术已应用于临床。在这个提案中我们旨在通过提出一种新颖的方法来解决这个问题经过基因校正的 iPSC 携带少量体细胞突变。我们相信我们的建议将对基于 iPSC 的安全性产生根本性影响疗法，并将作为制定释放标准的范例将用于鉴定 iPSC 库是否有任何遗传性和退行性疾病的资格。