Single Step Strategy for the Therapeutic Reprogramming in Epidermolysis Bullosa Patients

大疱性表皮松解症患者治疗性重编程的一步策略

基本信息

批准号：
9475196
负责人：
Vittorio Sebastiano
金额：
$ 17.27万
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/9475196
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 Genetic Diseases Genome Genomic DNA Goals Guide RNA 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.

使用人类诱导的多能干细胞（IPSC）进行再生药物是治疗遗传和退化的有吸引力的替代品大量或连续的组织再生需要的疾病是必需的。 IPSC更容易适应原位基因校正，可以是无限期保留多能和未分化状态的扩大，可以因此，用作细胞治疗的恒定材料来源。发展基于IPSC的遗传性营养不良表皮溶液（RDEB）的疗法在这种情况下，患者代表了理想的范例在该疾病中，校正角质形成细胞的演示可以长期组织重生，需要大量干细胞覆盖受影响的表面积。我们最近证明了治疗性重新编程（TR）（通过遗传校正RDEB患者来实现 IPSC）几乎提供了一个无限的遗传校正自体源的来源可以从具有恢复功能和的多层皮肤的角质形成细胞 1型胶原蛋白VII的生理表达（Sebastiano等，2014）。尽管有效的，这种方法的主要瓶颈之一是许多亚键盘得出校正后的IPSC克隆所需的步骤，并具有固有的积累大量的体细胞突变和核型异常（在与现场的明显证据体系达成协议）移植后体内不可预测的影响，并具有严重的影响关于一般基于IPSC的治疗的安全性。这代表了一个基本问题和杰出的优先级需要在翻译之前解决建立了基于IPSC的技术到诊所的技术。在这个建议中，我们是旨在通过提出一种新颖的方法来解决这个问题经遗传校正的IPSC携带纤细的体细胞突变。我们相信我们的建议将对基于IPSC的安全产生根本的影响疗法，将作为制定发布标准的范式将使用任何遗传和退化性疾病的IPSC银行资格。