The Role of Macrophages in Pulmonary Regeneration using a Bioengineered Whole Lung Tissue Model

使用生物工程全肺组织模型研究巨噬细胞在肺再生中的作用

• 批准号: 10387664
• 负责人: Allison Marie Greaney
• 金额: $ 6.72万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10387664
• 关键词: Air; Alveolar; Alveolar Cell; Alveolar Macrophages; Anti-Inflammatory Agents; Apoptotic; Architecture; Basal Cell; Behavior; Benchmarking; Biological Models; Biomechanics; Biomedical Engineering; Biomimetics; Bronchoalveolar Lavage; COVID-19; Cell Communication; Cells; Cessation of life; Chemicals; Coculture Techniques; Communication; Data; Dinoprostone; Disease; Disease Outcome; Distal; Endothelium; Engineering; Environment; Epithelial; Epithelial Cells; Evaluation; Faculty; Fibroblasts; Fibrosis; Goals; Histologic; Homeostasis; Human; Hypoxemia; In Vitro; Inflammatory; Injury; Institutes; Institution; Interferon Type II; Interferons; Interleukin-4; Intervention; Irrigation; Lead; Lipopolysaccharides; Liquid substance; Lung; Lung diseases; Maintenance; Massachusetts; Mentors; Mesenchymal; Mesenchyme; Methods; Muscle; Myoblasts; Myofibroblast; Natural regeneration; Paracrine Communication; Pattern; Pharmacology; Phenotype; Play; Population; Positioning Attribute; Production; Proteins; Protocols documentation; Publishing; Rattus; Reporting; Research; Research Project Grants; Role; Route; SARS-CoV-2 infection; Secure; Signal Transduction; Site; Skeletal muscle injury; Structure of parenchyma of lung; System; Technology; Therapeutic Intervention; Tissue Model; Tissues; Training; Universities; Vascular Endothelium; Work; alveolar epithelium; anticancer research; base; cell community; cell stroma; cell type; cytokine; epithelial repair; epithelial stem cell; epithelium regeneration; fibrotic lung; fibrotic lung disease; idiopathic pulmonary fibrosis; improved; in vivo; influenza infection; insight; lung regeneration; lung repair; macrophage; novel; organ growth; progenitor; recruit; regenerative; repaired; response; restoration; scaffold; severe injury; single-cell RNA sequencing; skills; stem cells; tenure track

PROJECT SUMMARY/ABSTRACT Fibrotic lung remodeling is a hallmark of many lung diseases, including severe influenza infection, idiopathic pulmonary fibrosis, and COVID-19. While a basal-like epithelial cell has been identified as a central player to this aberrant repair response, little has been done to investigate the behavior and interactions of other cells in the diseased tissue, particularly alveolar macrophages. Therefore, the goal of this study is to leverage a biomimetic engineered lung model system to investigate the relationship between regenerating basal-like progenitor cells and pulmonary macrophages. This engineered lung tissue system is based on cellular repopulation and culture of a decellularized native rat lung scaffold. This platform enables a well-controlled, native-like tissue environment for the evaluation of cell-cell interactions, without the systemic confounders of in vivo studies. It is expected that macrophages will significantly influence and direct epithelial remodeling by these basal-like cells, particularly in relation to the fibrotic or anti-fibrotic activation state of the macrophages. This work expands on previous findings that adding macrophages to engineered lung cultures containing basal-like progenitor cells significantly improves tissue architecture and regenerative epithelial cell phenotype, compared to engineered lung cultures without macrophages. First, pulmonary macrophages will be isolated from rats by bronchoavleolar lavage and characterized. Protocols will be developed to chemically stimulate macrophages in vitro to a disease-like inflammatory state, or to a reparative anti-fibrotic state. Next, macrophages of different activation states will be introduced to air-liquid interface cultures of regenerative basal cells, to evaluate epithelial-macrophage interactions in isolation. Finally, activated macrophages will be introduced to engineered lung cultures containing regenerative basal cells, fibroblasts, and endothelium to recapitulate essential native cellular communities. Engineered lung tissues will be evaluated for histologic and biomechanic changes between conditions, as well as differential cell signaling patterns, as evaluated by single-cell RNA sequencing. It is expected that inflammatory macrophages will contribute to fibrotic response in regenerating epithelium, whereas anti-fibrotic macrophages will contribute to more functional alveolar regeneration by basal-like cells. The findings of this study will elucidate the role of pulmonary macrophages in governing lung repair and regeneration in this model system. Further, this work may suggest possible routes for the treatment of fibrotic lung diseases. The proposed research project will be executed by Allison M. Greaney at the David H. Koch Institute for Integrative Cancer Research at the Massachusetts Institute of Technology (MIT), under the Sponsorship of Dr. Robert Langer, and Co-Sponsorship of Dr. Ruslan Medzhitov at Yale University. Dr. Langer and Dr. Medzhitov will mentor Allison in her Research and Professional Training Goals to develop new research and scientific communication skills, so she may be well-equipped to secure a tenure-track faculty position at a top research institution.

项目总结/摘要 纤维化肺重塑是许多肺部疾病的标志，包括严重的流感感染、特发性肺炎、呼吸道感染、 肺纤维化和新冠肺炎虽然基底样上皮细胞已被确定为这一过程的核心参与者， 异常的修复反应，很少有人做调查的行为和其他细胞的相互作用，在 病变组织，特别是肺泡巨噬细胞。因此，本研究的目标是利用仿生 工程化肺模型系统，以研究再生基底样祖细胞与 和肺巨噬细胞。这种工程肺组织系统是基于细胞再生和培养 脱细胞的原生大鼠肺支架。该平台能够实现良好控制的、类似天然的组织环境 用于评价细胞-细胞相互作用，而不存在体内研究的系统性混杂因素。预计在 巨噬细胞将显著影响和指导这些基底样细胞的上皮重塑，特别是在 与巨噬细胞的纤维化或抗纤维化活化状态有关。这项工作扩展了以前的发现 在含有基底样祖细胞的工程肺培养物中加入巨噬细胞， 组织结构和再生上皮细胞表型，与无 巨噬细胞首先，通过支气管肺泡灌洗从大鼠中分离肺巨噬细胞， 表征了将制定方案，在体外以化学方式刺激巨噬细胞， 炎症状态或修复性抗纤维化状态。接下来，不同激活状态的巨噬细胞将被 引入再生基底细胞的气-液界面培养，以评估上皮-巨噬细胞 孤立的互动。最后，将活化的巨噬细胞引入工程肺培养物中， 再生的基底细胞、成纤维细胞和内皮细胞，以重现基本的天然细胞群落。 工程肺组织将评估组织学和生物力学的变化之间的条件，以及 作为差异细胞信号模式，如通过单细胞RNA测序评估的。预计在 炎性巨噬细胞将有助于再生上皮中的纤维化反应，而抗纤维化巨噬细胞将有助于再生上皮中的纤维化反应。 巨噬细胞将通过基底样细胞促进更多功能性肺泡再生。这一发现 本研究将阐明肺巨噬细胞在该模型中调控肺修复和再生的作用 系统此外，这项工作可能为纤维化肺疾病的治疗提供可能的途径。拟议 研究项目将由Allison M.在大卫H的格里尼。科赫综合癌症研究所 在马萨诸塞州理工学院（MIT）的研究，由Robert Langer博士赞助， 耶鲁大学鲁斯兰·梅德日托夫博士的共同赞助。兰格博士和梅兹托夫博士将指导艾莉森 她的研究和专业培训目标，以发展新的研究和科学沟通技能， 她也许有能力在顶尖研究机构获得终身教职。