iPSC-Derived Alveolar Epithelial Cells for Intrapulmonary Therapies

 描述(申请人提供)：本申请是根据PAR-13-095提出的，目的是测试来源于人类诱导多能干细胞(HiPSC)的谱系定向细胞，用于治疗先天性肺部疾病。这项提案中概述的方法建立在我们团队在发育个体发育、组织工程、干细胞/祖细胞研究、新颖的活体成像模式和翻译的非人类灵长类动物模型方面的专业知识的基础上。我们研究了将人类多能细胞分化为与肺相关的细胞系的方法，包括呼吸道上皮细胞(AECs)；利用独特的支架和基质构建了工程结构；并开发了有效标记、移植和监测猕猴体内移植前体命运的技术。我们现在准备推进我们的先前发现，并开辟新的领域，通过以下具体目标解决与IPSC来源的前体细胞相关的关键问题，以治疗先天性肺疾病，如肺表面活性物质缺乏：(1)优化HiPSCs在3D培养中的传代承诺，以提高AEC的分化和纯度；以及(2)进行概念验证研究，以评估胎儿移植效率和细胞在体内的命运。我们将在3D培养中优化关键细胞群的增殖和整合，以提高谱系规格，并通过包括生物发光成像和正电子发射断层扫描/计算机断层扫描(PET/CT)在内的创新成像范式评估移植效率和体内细胞命运。肺的修复能力有限，基于发育原则的再生概念，根据患者的年龄量身定做，在翻译灵长类动物模型中进行测试，并使用最先进的成像技术，可能会对该领域产生重大影响。这些调查将提供新的见解和指导转化研究的新途径，这些研究可能会对儿童和所有年龄段的人产生重大影响和令人信服的好处。

 DESCRIPTION (provided by applicant): This application is submitted in response to PAR-13-095 with the intent of testing lineage committed cells derived from human induced pluripotent stem cells (hiPSC) for the treatment of congenital lung diseases. The approaches outlined in this proposal build on the expertise of our team in developmental ontogeny, tissue engineering, stem/progenitor cell studies, novel in vivo imaging modalities, and a translational nonhuman primate model. We have studied methods to differentiate human pluripotent cells towards lung-related lineages including airway epithelial cells (AECs); engineered constructs with unique scaffolds and matrices; and developed techniques to effectively label, transplant, and monitor the fate of transplanted precursors in vivo in rhesus monkeys. We are now poised to advance our prior discoveries and break new ground to address key questions related to iPSC-derived precursors to treat congenital lung diseases, such as pulmonary surfactant deficiency, through the following Specific Aims: (1) Optimize lineage commitment of hiPSCs in 3D culture to enhance AEC differentiation and purity, and (2) Conduct proof-of-concept studies to evaluate fetal transplant efficiency and cell fate in vivo. We will optimize proliferation and integration o key cell populations in 3D culture to enhance lineage specification, and evaluate transplant efficiency and cell fate in vivo through innovative imaging paradigms including bioluminescence imaging and positron emission tomography/computed tomography (PET/CT). The lung has limited capacity for repair, and regenerative concepts that are based on developmental principles, tailored to the age of the patient, tested in a translational primate model, and use state-of-the-art imaging technologies could have a major impact on the field. These investigations will provide new insights and guide new avenues of translational research that could have significant impact and compelling benefits for children and all age groups across the lifespan.