Gene corrected iPS-derived lung cells for therapy of inherited lung diseases

描述（由申请人提供）：肺干/祖细胞的细胞移植代表了多种遗传性肺病的潜在治疗方法。这种治疗策略成功的关键是移植的细胞及其后代针对致病突变进行校正（基因型和表型），并且移植的细胞不会在接受者中引起免疫应答。最近开发的用于产生诱导的多能干（iPS）细胞和锌指核酸酶（ZFN）介导的基因组编辑的方法原则上使得：B）特异性校正自体iPS细胞的染色体DNA中的责任基因突变，和c）iPS细胞体外分化为细胞移植治疗所需的各种细胞类型。我们建议采用这种方法为患有表面活性蛋白B（SP-B）缺乏症或囊性纤维化（CF）的患者产生校正的自体iPS细胞。对于SP-B缺乏，我们将进一步产生具有对患有这种疾病的患者进行细胞治疗的潜力的校正的肺祖细胞（肺泡上皮II型细胞; ATII）。这个潜在的高影响力的两年期项目汇集了两个高素质的主要研究人员的互补专业知识：一个（布赖恩R。Davis）在iPS细胞的产生/表征以及ZFN介导的基因组编辑方面有经验，另一个（Rick A. Wetsel）在人多能干细胞体外定向分化为肺祖细胞和将这种祖细胞移植到小鼠肺中方面经验丰富。这种新的方法学的融合代表了适用于其他单基因引起的肺、心脏和血液系统疾病的细胞治疗的范例。 公共卫生相关性：本项目的目的是产生校正的、患者特异性的肺祖细胞，用于遗传性肺病患者的细胞移植治疗。它代表了从体细胞产生自体多能干细胞、染色体DNA中基因突变的有效位点特异性校正以及多能细胞定向分化为适于移植的细胞类型的方法学的新融合。

DESCRIPTION (provided by applicant): Cellular transplantation of lung stem/progenitor cells represents a potential therapeutic approach for a variety of inherited lung diseases. Crucial to the success of such a therapeutic strategy is that the transplanted cells and their progeny are corrected (both genotypically and phenotypically) for the disease-causing mutation and that the transplanted cells do not elicit an immune response in the recipient. Recently developed methodologies for generating induced pluripotent stem (iPS) cells and zinc finger nuclease (ZFN)-mediated genome editing make possible, in principle: a) the generation of autologous, patient-specific iPS cells carrying inherited genetic mutations; b) specific correction of the responsible genetic mutation in chromosomal DNA of the autologous iPS cells, and c) in vitro differentiation of iPS cells into various cell types required for cell transplantation therapy. We propose to employ this approach to generate corrected, autologous iPS cells for patients with either Surfactant Protein B (SP-B) Deficiency or Cystic Fibrosis (CF). For SP-B deficiency we will further generate corrected lung progenitor cells (lung alveolar epithelial type II cells; ATII) with the potential for cellular therapy of patients with this disease. This potentially high-impact two year project brings together the complementary expertise of two highly qualified principal investigators: one (Brian R. Davis) experienced in the generation/characterization of iPS cells together with ZFN-mediated genome editing, the other (Rick A. Wetsel) experienced in directed, in vitro differentiation of human pluripotent stem cells to lung progenitor cells and transplantation of such progenitor cells into mouse lung. This novel convergence of methodologies represents a paradigm applicable to cellular therapy of other single-gene caused diseases of the lung, heart, and blood systems. PUBLIC HEALTH RELEVANCE: The objective of this project is to generate corrected, patient-specific lung progenitor cells for cell transplantation therapy of patients with inherited lung disease. It represents a novel convergence of methodologies for generation of autologous, pluripotent stem cells from somatic cells, efficient site-specific correction of genetic mutations in chromosomal DNA, and directed differentiation of pluripotent cells to cell types appropriate for transplantation.