Alveolar Epithelial Cell Dysfunction in Pulmonary Fibrosis: Leveraging SFTPC Mutations for Discovery of Molecular and Cellular Targets

肺纤维化中的肺泡上皮细胞功能障碍：利用 SFTPC 突变发现分子和细胞靶点

• 批准号: 10407546
• 负责人: MICHAEL FRANCIS BEERS
• 金额: $ 55.15万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407546
• 关键词: Adopted; Adult; Affect; Alleles; Alveolar; Alveolitis; Appearance; Architecture; Biological Markers; CCL2 gene; Cell Communication; Cells; Cessation of life; Child; Cicatrix; Clinical; Codon Nucleotides; Complex; Complication; Consensus; Data; Development; Distal; Epithelial; Epithelial Cells; Event; Evolution; Fibroblasts; Fibrosis; Functional disorder; Funding; Gases; Gene Expression; Genes; Genetic; Genetic Models; Genetic Transcription; Growth; Hand; Homeostasis; Human; In Vitro; Inflammatory; Injury; Interstitial Lung Diseases; Kinetics; Knock-in Mouse; Ligands; Link; Lung; Mediator of activation protein; Medical; Mesenchymal; Modeling; Molecular; Molecular Conformation; Molecular Target; Mus; Mutation; Organelles; Organoids; Outcome; Paracrine Communication; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Peripheral; Phase; Phenotype; Physiological; Population; Pre-Clinical Model; Process; Production; Protein Isoforms; Proteins; Publishing; Pulmonary Fibrosis; Pulmonary Surfactant-Associated Protein C; Reagent; Reporting; Respiratory Failure; Role; Signal Pathway; Signal Transduction; Testing; Therapeutic; Time; Tissues; alveolar epithelium; base; cellular targeting; cysteinylglycine; cytokine; endoplasmic reticulum stress; fibrotic interstitial lung disease; fibrotic lung; fibrotic lung disease; genetic variant; in vitro Model; in vivo; insight; lung injury; monocyte; mouse model; multimodality; mutant; non-Native; novel; older patient; progenitor; programs; receptor; recruit; response; stem cell function; stem cells; tool; transcriptomics; wound healing; young adult

ABSTRACT Fibrotic lung remodeling represents the final common pathway to respiratory failure for a variety of Interstitial Lung Diseases (ILD) in children and adults. Based on a recent paradigm shift wherein the concepts of epithelial cell dysfunction and abnormal wound healing are postulated as “drivers” of pulmonary fibrosis, new opportunities are emerging for therapeutic discovery for ILD. Mutations in the Surfactant Protein C [SP-C] gene [SFTPC], an alveolar type 2 cell (AT2) restricted protein, have been found in sporadic and familial ILD and can provide important clues for understanding the role of epithelial cell dysfunction in ILD pathogenesis. Prior in vitro studies from our lab have shown that SFTPC mutations described in both adults and children with ILD result in production of aberrant SP-C proprotein isoforms that adopt non-native conformations resulting in at least 2 outcomes: ER stress and intracellular aggregation (BRICHOS) or mistrafficking to non-native organelles and inhibition of macroautophagy (Non-BRICHOS). This application proposes to leverage two novel knock-in mouse models of spontaneous lung fibrosis already in hand which express Non-BRICHOS or BRICHOS clinical SFTPC mutants in AT2 cells in an allelic and inducible fashion. Our Published and Preliminary Data reveal that expression of either a non-BRICHOS mutant (SP-CI73T) or BRICHOS mutant (SP- CC121G) is extremely toxic to the lung in vivo with each resulting in time-dependent spontaneous lung fibrosis marked by 3 phases: ii) early AT2 cytokine elaboration and monocyte recruitment,; (ii) a polycellular alveolitis and lung injury; (iii) physiologically restrictive peripheral fibrotic remodeling. These models also elaborate translationally relevant biomarkers reported in human ILD. In order to define both consensus and divergent molecular mechanisms linking these two AT2 cell phenotypes with the downstream lung injury and fibrotic lung remodeling, our experimental approach will be to exploit the unique features of these genetic models combined with tools, reagents, and expertise available in our program. In 3 specific aims, we propose to comprehensively characterize the transcriptomic and functional AT2 cell phenotypes evoked by expression of non-BRICHOS (SP-CI73T) and BRICHOS (SP-CC121G) Sftpc mutants [Specific Aim 1], to define the role of a key proinflammatory/profibrotic monocyte population recruited by AT2 mutant Sftpc expression in the development of fibrosis [Specific Aim 2], and to examine changes in epithelial-mesenchymal crosstalk that disrupt alveolar niche homeostasis and promote fibrotic remodeling [Specific Aim 3]. As epithelial dysfunction has not been studied extensively in vivo in the context of relevant preclinical models of fibrotic lung disease, this approach offers the unique opportunity to provide proof of concept both for the causal effect of mutant SFTPC in familial ILD and for the role of AT2 dysfunction as a key upstream driver of inflammatory cell and fibroblast activities that promote parenchymal remodeling. Importantly, mechanisms identified using a focused approach with these Sftpc models can be cross-purposed to better understand the pathogenesis of sporadic forms of ILD.

抽象的 纤维化肺重塑代表了多种间隙的最终呼吸衰竭途径 儿童和成人的肺部疾病（ILD）。基于最近的范式转变 上皮细胞功能障碍和异常伤口愈合被张贴为肺纤维化的“驱动因素”，新的 为ILD提供治疗发现的机会正在出现。表面活性剂蛋白C [sp-c]的突变 在零星和家族ILD中发现了基因[SFTPC]，这是一种大型2型细胞（AT2）受限蛋白 并可以提供重要的线索，以了解上皮细胞功能障碍在ILD发病机理中的作用。 我们实验室的先前体外研究表明，成人和儿童中描述的SFTPC突变 ILD导致产生异常的SP-C proprotein同工型，这些同工型采用非母构构，导致 至少有2个结果：ER应力和细胞内聚集（Brichos）或不舒服到非本地 细胞器和大型噬菌体的抑制（非砖）。该申请提案要利用两个 新颖的敲入肺肺纤维化的小鼠模型已经表达非砖头或 AT2细胞中的Brichos临床SFTPC突变体以等位基因和可诱导的方式。我们出版的 初步数据表明，非砖头突变体（SP-CI73T）或Brichos突变体（SP-）的表达 CC121G）对体内的肺极具毒性，每种都会导致时间依赖性的赞助肺纤维化 以3个阶段为单位：ii）AT2早期的AT2细胞因子阐述和单核细胞募集； （ii）多细胞肺泡炎 和肺部受伤； （iii）物理限制性的外周纤维化重塑。这些模型也详细说明 人类ILD报道了翻译相关的生物标志物。为了定义共识和分歧 将这两个AT2细胞表型与下游肺损伤和纤维化肺部联系起来的分子机制 重塑，我们的实验方法是探索这些遗传模型的独特特征 使用我们的计划中提供的工具，试剂和专业知识。在3个特定目标中，我们建议全面 表征非砖头表达引起的转录组和功能性AT2细胞表型 （SP-CI73T）和Brichos（SP-CC121G）SFTPC突变体[特定目标1]，以定义钥匙的作用 在开发中，AT2突变体SFTPC募集的促炎/纤维化单核细胞种群 纤维化[特定目标2]，并检查上皮 - 间质串扰的变化，这些变化破坏了肺泡 利基稳态并促进纤维化重塑[特定目标3]。由于上皮功能障碍尚未 在相关的纤维化肺疾病的临床前模型中，在体内进行了广泛的研究，这种方法 提供了独特的机会，可以为突变体SFTPC的因果效应提供概念证明 ILD和AT2功能障碍作为炎症细胞和成纤维细胞活动的关键上游驱动力的作用 这促进了副型重塑。重要的是，使用集中方法确定的机制 这些SFTPC模型可以交叉插入以更好地理解ILD的零星形式的发病机理。

项目成果

Microstructured Hydrogels to Guide Self-Assembly and Function of Lung Alveolospheres.

• DOI: 10.1002/adma.202202992
• 发表时间: 2022-07
• 期刊: ADVANCED MATERIALS
• 影响因子: 29.4
• 作者: Loebel, Claudia;Weiner, Aaron, I;Eiken, Madeline K.;Katzen, Jeremy B.;Morley, Michael P.;Bala, Vikram;Cardenas-Diaz, Fabian L.;Davidson, Matthew D.;Shiraishi, Kazushige;Basil, Maria C.;Ferguson, Laura T.;Spence, Jason R.;Ochs, Matthias;Beers, Michael F.;Morrisey, Edward E.;Vaughan, Andrew E.;Burdick, Jason A.
• 通讯作者: Burdick, Jason A.

Integrated plasma proteomics and lung transcriptomics reveal novel biomarkers in idiopathic pulmonary fibrosis.

• DOI: 10.1186/s12931-021-01860-3
• 发表时间: 2021-10-24
• 期刊: Respiratory research
• 影响因子: 5.8
• 作者: Sivakumar P;Ammar R;Thompson JR;Luo Y;Streltsov D;Porteous M;McCoubrey C;Cantu E 3rd;Beers MF;Jarai G;Christie JD
• 通讯作者: Christie JD

Chronic Expression of a Clinical SFTPC Mutation Causes Murine Lung Fibrosis with Idiopathic Pulmonary Fibrosis Features.

临床 SFTPC 突变的慢性表达导致具有特发性肺纤维化特征的小鼠肺纤维化。

• DOI: 10.1165/rcmb.2022-0203ma
• 发表时间: 2023
• 期刊: American journal of respiratory cell and molecular biology
• 影响因子: 6.4
• 作者: Rodriguez,Luis;Tomer,Yaniv;Carson,Paige;Dimopoulos,Thalia;Zhao,Ming;Chavez,Katrina;Iyer,Swati;Huang,Li;Ebert,Christina;Sereda,Larisa;Murthy,Aditi;Trujillo,Glenda;Beers,MichaelF;Katzen,Jeremy
• 通讯作者: Katzen,Jeremy

Immunophenotyping of Acute Inflammatory Exacerbations of Lung Injury Driven by Mutant Surfactant Protein-C: A Role for Inflammatory Eosinophils.

• DOI: 10.3389/fphar.2022.875887
• 发表时间: 2022
• 期刊: FRONTIERS IN PHARMACOLOGY
• 影响因子: 5.6
• 作者: Nguyen, Jacklyn;Armstrong, Brittnie S.;Cowman, Sophie;Tomer, Yaniv;Veerabhadraiah, Shivakumar R.;Beers, Michael F.;Venosa, Alessandro
• 通讯作者: Venosa, Alessandro

STAT3-BDNF-TrkB signalling promotes alveolar epithelial regeneration after lung injury.

• DOI: 10.1038/s41556-020-0569-x
• 发表时间: 2020-10
• 期刊: Nature cell biology
• 影响因子: 21.3
• 作者: Paris AJ;Hayer KE;Oved JH;Avgousti DC;Toulmin SA;Zepp JA;Zacharias WJ;Katzen JB;Basil MC;Kremp MM;Slamowitz AR;Jayachandran S;Sivakumar A;Dai N;Wang P;Frank DB;Eisenlohr LC;Cantu E 3rd;Beers MF;Weitzman MD;Morrisey EE;Worthen GS
• 通讯作者: Worthen GS