Utilizing induced pluripotent stem cells to study the role of alveolar type 2 cell dysfunction in pulmonary fibrosis

项目摘要 该提案详细介绍了一项为期5年的职业发展培训计划，重点是开发针对患者的 诱导多能干细胞（iPSC）模型系统，以研究肺泡上皮2型（AT 2）细胞的作用 肺纤维化（PF）初期的功能障碍。越来越多的文献认为肺泡上皮细胞 功能障碍在导致下游成纤维细胞活化的事件中发挥作用， 各种间质性肺病（ILD）综合征中的纤维化，包括成人特发性PF（IPF）和儿童 ILD（儿童）。然而，由于没有获得患者特异性的人上皮-间充质模型系统， 对于测试AT 2细胞功能障碍如何导致人类疾病的假设，选择有限。概述的 该提案建立在候选人最近开发和发表的体外人体模型系统的基础上， 了解AT 2细胞功能障碍在最常见疾病中的机制- 相关的SFTPC变体（SFTPCI 73 T）导致PF。通过研究AT 2细胞功能障碍来确定其机制 将在SftpcI 73 T突变小鼠中进一步体内验证使用体外iPSC衍生模型的方法。更具体地说， 本提案的目的是：1）研究AT 2细胞功能障碍的作用和下游后果， 体外利用新型人类患者特异性iPSC诱导纤维化级联反应中的上皮功能障碍 上皮-间充质重组模型系统，2）检验AT 2细胞-内源性扰动 其特征在于蛋白质稳态缺陷和代谢重编程导致受损的AT 2-to-AT 1细胞 分化、炎性活化和纤维化间充质活化，以及3）鉴定可药物化的 通过测试恢复AT 2细胞蛋白质稳态和线粒体的新方法， 功能将要开发的模型系统和将要揭示的致病机制都可能是 可推广到广泛多样的PF表型，为家族性和 偶发性PF治疗。Alysandratos有80%的保护时间从肺，过敏，睡眠司 和重症监护医学和波士顿大学医学系。他的导师达雷尔·克顿博士在 再生医学中心（CReM）是干细胞生物学的国际专家，专注于 将干细胞应用于建模并了解肺部发育和疾病，使他非常适合这项研究。 职业发展奖专注于PF的iPSC模型系统。一个非凡的科学顾问团队 各成员都带来了各自的专门知识，经过精心召集， 指导一个详细的培训计划，包括指导研究，教学课程， 会议上的演讲，完成研究目标的时间轴，准备手稿，以及 未来的R 01应用。在完成这一建议时，候选人将制定必要的 专业知识，成功地过渡到一个独立的医生，科学家。

Project Summary This proposal details a 5-year career development training program focused on developing a patient-specific induced pluripotent stem cell (iPSC) model system to study the role of alveolar epithelial type 2 (AT2) cell dysfunction at the inception of pulmonary fibrosis (PF). A growing literature now implicates alveolar epithelial dysfunction as playing a role in the events that lead to downstream fibroblast activation culminating in relentless fibrosis in a variety of interstitial lung disease (ILD) syndromes, including adult idiopathic PF (IPF) and childhood ILD (chILD). However, without access to patient-specific human epithelial-mesenchymal model systems, there are limited options for testing hypotheses of how AT2 cell dysfunction leads to disease in humans. The outlined proposal builds on an in vitro human model system recently developed and published by the candidate to better understand the mechanisms by which AT2 cell dysfunction in the context of the most common disease- associated SFTPC variant (SFTPCI73T) leads to PF. The mechanisms identified by studying AT2 cell dysfunction using the in vitro iPSC-derived model will be further validated in vivo in SftpcI73T mutant mice. More specifically, the aims of this proposal are to: 1) study the role of AT2 cell dysfunction and the downstream consequences of epithelial dysfunction in eliciting a fibrotic cascade by utilizing a novel human patient-specific iPSC in vitro epithelial-mesenchymal recombinant model system, 2) test the hypothesis that AT2 cell-intrinsic perturbations characterized by proteostasis defects and metabolic reprograming result in impaired AT2-to-AT1 cell differentiation, inflammatory activation, and fibrogenic mesenchymal activation, and 3) identify druggable pathways for novel PF therapies by testing novel approaches to restore AT2 cell proteostasis and mitochondrial function. Both the model system to be developed and the pathogenic mechanisms to be revealed likely will be generalizable to a broad diversity of PF phenotypes, providing novel druggable targets for both familial and sporadic PF therapies. Dr. Alysandratos has 80% protected time from the Division of Pulmonary, Allergy, Sleep & Critical Care Medicine and the Boston University Department of Medicine. His mentor, Dr. Darrell Kotton at the Center for Regenerative Medicine (CReM), is an international expert in stem cell biology with a focus on applying stem cells to model and understand lung development and disease, making him ideally suited for this career development award focused on iPSC-model systems of PF. A team of extraordinary scientific advisory members, each bringing their specific expertise, has been carefully assembled to provide complementary guidance. A detailed training plan is presented that includes mentored research, didactic coursework, presentations at meetings, and a timeline for completion of the research aims, preparation of manuscripts, and future R01 application. At the completion of this proposal, the candidate will have developed the necessary expertise to successfully transition into an independent physician-scientist.