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

描述(申请人提供)：肺干细胞/祖细胞的细胞移植代表了一种潜在的治疗各种遗传性肺部疾病的方法。这种治疗策略成功的关键是移植的细胞及其后代被纠正(包括基因和表型)以纠正致病突变，并且移植的细胞不会在接受者中引发免疫反应。最近开发的产生诱导多能干细胞(IPS)的方法和锌指核酸酶(ZFN)介导的基因组编辑原则上使以下方面成为可能：a)产生携带遗传突变的自体、患者特有的iPS细胞；b)特异地纠正自体iPS细胞染色体DNA中负责的基因突变；以及c)体外将iPS细胞分化为细胞移植治疗所需的各种细胞类型。我们建议使用这种方法为表面活性蛋白B(SP-B)缺乏或囊性纤维化(CF)的患者生成校正的、自体的iPS细胞。对于SP-B缺乏，我们将进一步培养纠正的肺祖细胞(肺泡上皮II型细胞；ATII)，为这种疾病的患者提供细胞治疗的可能性。这个潜在的高影响力项目为期两年，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细胞的产生/表征以及ZFN介导的基因组编辑方面经验丰富，另一位(Rick A.Wetsel)在人类多能干细胞体外定向分化为肺祖细胞并将此类祖细胞移植到小鼠肺内方面经验丰富。这种新的方法学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的范例。 公共卫生相关性：该项目的目标是为遗传性肺部疾病患者的细胞移植治疗产生校正的、患者特有的肺祖细胞。它代表了一种新的融合方法，用于从体细胞生成自体多能干细胞，有效地定点纠正染色体DNA的遗传突变，并将多能细胞定向分化为适合移植的细胞类型。

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.