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部疾病的潜在治疗方法。这种治疗策略成功的关键在于，移植细胞及其后代对致病突变进行了纠正（包括基因型和表型），并且移植细胞不会引起受体的免疫反应。最近开发的诱导多能干细胞（iPS）和锌指核酸酶（ZFN）介导的基因组编辑方法，原则上使：a)产生携带遗传基因突变的自体、患者特异性iPS细胞成为可能；b)特异性纠正自身iPS细胞染色体DNA的相关基因突变，c) iPS细胞在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表面活性剂蛋白B （SP-B）缺乏或囊性纤维化（CF）患者生成校正的自体iPS细胞。对于SP-B缺乏症，我们将进一步生成校正后的肺祖细胞（肺肺泡上皮II型细胞；ATII），有可能用于该疾病患者的细胞治疗。这个潜在的高影响力的为期两年的项目汇集了两位高素质的主要研究人员的互补专业知识：一位（Brian R. Davis）在iPS细胞的生成/表征以及zdn介导的基因组编辑方面经验丰富，另一位（Rick A. Wetsel）在人类多能干细胞定向体外分化为肺祖细胞并将这种祖细胞移植到小鼠肺中。这种方法的新颖融合代表了一种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范例。

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.