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

利用诱导多能干细胞研究肺泡2型细胞功能障碍在肺纤维化中的作用

• 批准号: 10591174
• 负责人: Konstantinos Alysandratos
• 金额: $ 16.22万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10591174
• 关键词: Adult; Alveolar; Biogenesis; Biological Models; Boston; Cell Differentiation process; Cell Line; Cells; Cessation of life; Childhood; Critical Care; Data; Data Set; Defect; Development; Diagnosis; Disease; Disease model; Epithelium; Event; FDA approved; Fibroblasts; Fibrosis; Functional disorder; Future; Gases; Genes; Genetic study; Genotype; Goals; Human; Hypersensitivity; Idiopathic Interstitial Pneumonia; Immune; Impairment; In Vitro; Inflammatory; International; Interstitial Lung Diseases; K-Series Research Career Programs; Literature; Lung; Lung diseases; Manuscripts; Medicine; Mentors; Mesenchymal; Mesenchyme; Metabolic; Methods; Mitochondria; Modeling; Morbidity - disease rate; Morphology; Mutant Strains Mice; Mutation; Organoids; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physicians; Pirfenidone; Pluripotent Stem Cells; Population; Preparation; Proteomics; Publishing; Pulmonary Fibrosis; Recombinants; Regenerative Medicine; Research; Role; Scientist; Sleep; Stress; Structure of parenchyma of lung; Syndrome; Systems Biology; Testing; Time; Training; Training Programs; Universities; Variant; alveolar epithelium; career development; disorder risk; effective therapy; fibrogenesis; genome wide association study; genomic locus; human disease; human model; idiopathic pulmonary fibrosis; in vitro Model; in vivo; in vivo evaluation; induced pluripotent stem cell; lung development; meetings; member; mitochondrial dysfunction; molecular phenotype; mortality; mouse model; new therapeutic target; nintedanib; novel; novel strategies; novel therapeutics; proteostasis; pulmonary function decline; self-renewal; single-cell RNA sequencing; stem cell biology; stem cell model; stem cells; timeline; transcriptomics

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.

项目摘要 这份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)模型系统研究肺泡上皮2型(AT2)细胞的作用 肺纤维化(PF)开始时的功能障碍。现在越来越多的文献表明肺泡上皮细胞 功能障碍在导致下游成纤维细胞激活的事件中发挥作用，最终导致无情 各种间质性肺疾病(ILD)综合征的纤维化，包括成人特发性肺纤维化(IPF)和儿童 ILD(儿童)。然而，由于无法访问特定于患者的人类上皮-间充质模型系统， 测试AT2细胞功能障碍如何导致人类疾病的假设的选择有限。概述 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，以更好地 了解AT2细胞功能障碍在最常见疾病背景下的机制- 相关的SFTPC变异体(SFTPCI73T)导致PF。AT2细胞功能障碍研究确定的机制 使用体外IPSC衍生模型将在SftpcI73T突变小鼠体内进一步验证。更确切地说， 这项建议的目的是：1)研究AT2细胞功能障碍的作用及其下游后果 利用一种新的人患者特异性IPSC在体外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AT2细胞内源性扰动假说 以蛋白平衡缺陷和代谢重编程为特征的AT2-to-AT1细胞受损 分化、炎症激活和纤维化间质激活，以及3)确定可用药 通过测试恢复AT2细胞蛋白平衡和线粒体的新方法为PF治疗提供新途径 功能。无论是要开发的模型系统，还是要揭示的致病机制都可能是 可推广到广泛多样化的Pf表型，为家族和 零星的PF疗法。AlySandratos博士有80%的保护时间来自肺，过敏，睡眠部门 和重症护理医学和波士顿大学医学部。他的导师，达雷尔·科顿博士 再生医学中心(CReM)是干细胞生物学的国际专家，专注于 应用干细胞来建模和了解肺发育和疾病，使他成为这方面的理想人选 职业发展奖聚焦于PF的IPSC模式系统。一支非凡的科学顾问团队 每个成员都带来了各自的专业知识，经过精心组装，以提供互补的 指导。提出了详细的培训计划，包括有指导的研究、授课课程、 在会议上的陈述，以及完成研究目标、准备手稿的时间表，以及 未来的R01应用。在完成这项建议时，候选人将制定出必要的 成功转型为独立内科医生-科学家的专业知识。